The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria and Bifidobacteria

*Ewa Kowalska and Małgorzata Ziarno*

## **Abstract**

In this study, we aimed to examine the effect of four different industrial starter cultures containing lactic acid bacteria and bifidobacteria on the selected characteristics of beverages prepared from buckwheat and stored at 4°C for 28 days. We estimated the pH of the beverages during fermentation and storage under refrigerated conditions. We also determined the number of lactic acid bacteria and bifidobacteria and performed a chromatographic analysis of the carbohydrates. According to the results, the tested starter cultures effectively fermented the buckwheat beverage. The viable cell count of the starter microflora was sufficient to demonstrate the health-promoting properties of buckwheat. The pH of beverages was stable during the refrigerated storage. However, the carbohydrate content of the stored beverages changed, which indicates a constant biochemical activity of the microflora.

**Keywords:** buckwheat, health, lactic acid bacteria, lactic acid fermentation, bifidobacteria, probiotics

## **1. Introduction**

In recent years, the eating habits of people have changed dramatically due to various reasons. One such reason is consumer awareness of the impact of food on human health. Products that have a natural composition, that are unprocessed, and are nongenetically modified are preferred the most by the consumers. Another important factor, which determines people's eating habits, is food allergies and intolerances, which eliminate the possibility of consumption of a particular food product. Food allergies and metabolic disorders have led to an increased demand for allergen-free food products that meet the daily requirements for protein and other nutrients. For example, in the case of gluten intolerance, it is impossible to eat food products containing gluten. For such individuals, an alternative food product is, among others, buckwheat, which, as a gluten-free pseudocereal, can be used as groats, flour in baking bread or cakes.

Buckwheat has a rich composition and high nutritional value and can be an ideal base for products that are enriched with lactic acid bacteria (LAB), including probiotics. They are defined as a functional food because when they are administered in adequate amounts, they confer specific health benefits to the consumer. Consuming functional foods helps to reduce the risk of developing diseases of affluence, such as diabetes, obesity, or cancer. Buckwheat beverage enriched with LAB and bifidobacterial is one such functional food. Its unique taste and nutritional value might be utilized to develop a new product dedicated to people with disorders of the digestive system, as well as for people who want to stay healthy.

Fermented plant-based products represent a better way to substitute dairy products that cannot be consumed by people with food allergies or intolerance. The plant-based products gain a pro-health value after the process of fermentation and at the same time, they require minimal processing. Furthermore, the probiotic LAB have a positive effect on human health by regulating the functions of the intestinal microbiota. They keep the digestive system healthy and increase immunity. They have anticarcinogenic and antiallergenic effects [1]. Food intolerances are not related to the immune system; they are caused by sensitivity to certain food ingredients, e.g. gluten [2]. At present, approximately 20% of the population is affected due to food intolerance [3]. So far, the detailed mechanism of food intolerance is not known, but it may be related to the neuroendocrine system of the digestive system [4]. In the case of treatment available for gluten intolerance, elimination of gluten from the diet is recommended. Any amount of gluten might be harmful to individuals who are gluten-intolerant. According to the literature, more than 50–100 mg of gluten per day can prove to be harmful to such individuals [5].

## **2. Plant-based beverages as an alternative for dairy-based probiotic beverages**

Buckwheat is a dicotyledonous plant and is referred to as a pseudocereal. It is classified as a secondary plant. It has a tap root system, which is 1 meter long, and has a straight stem, 60–90 cm high, and brown in color; it bears pink or white flowers. Different products are made from different parts of the plant. The grains are used to produce buckwheat flour and buckwheat, while straw, after threshing the seeds, is added to various types of fodder. During the flowering season, buckwheat provides nectar to the bees [6, 7]. Buckwheat kernels contain glutenfree protein and have well-balanced amino acid profile. The flour is a rich source of minerals such as copper, zinc, manganese, potassium, magnesium, phosphorus, and potassium. It is also rich in polyphenols such as rutin, orientin, vitexin, quercetin, isovitoxin, and isoorientin. Among the aforementioned polyphenols, rutin shows the strongest anti-inflammatory, anticancer, and protective effect. In terms of flavonoid content, tartar buckwheat seeds contain approximately 40 mg/g, whereas common buckwheat seeds contain 10 mg/g [8–10]. A previous study [11] reported that sucrose is the predominant sugar in buckwheat, whereas xylose, glucose, arabinose, and melibiose are present in much smaller quantities. According to a previous study [12], sucrose accumulates in large quantities when the dry matter content is increased. It mainly occurs in the central part of the ovule and the seed coat. Buckwheat also contains *R*-tocopherol, which shows vitamin E activity [13]. In addition, the ethanolic extract of buckwheat contains four catechins: epicatechin, catechin 7-O-β-D-glucopyranoside, (−)-epicatechin 3-O-p-hydroxybenzoate, and (−)-epicatechin-3-O-(3,4-di-O) gallate-methyl [13]. It should be noted that the content of individual components in the plant may change depending on the environmental factors such as temperature, UV radiation, and damage caused by pests. Genetic factors are also of great importance, and the influence of the height of cultivation to sea level has been recently demonstrated [14].

**79**

*The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria…*

they produce mucus; and finally, they synthesize B vitamins.

are polyphenols, sterols, carotenoids, probiotics, and prebiotics [21].

are removed to avoid the aggravation of the disease [22].

as yogurt reduces the risk of heart disease and type 2 diabetes [26, 27].

The word "probiotic" was borrowed from Greek, wherein "probios" means "for life." Probiotics are mainly bacterial strains from the genera *Lactobacillus* and *Bifidobacterium*. However, before a strain is considered a probiotic, clinical trials must be conducted to prove its health-promoting properties [15, 16]. There should be mutual benefit between the human body and the probiotic bacteria (on the basis of symbiosis). The intestines are one of the most important organs in maintaining the body's normal immunity. About 70% of the entire population of immune cells is associated with intestinal mucosa [1]. Literature shows that probiotics regulate the functioning of the bacterial microflora in the intestines through certain mechanisms [17]; for example, they compete with the pathogenic bacteria for the same receptors and nutrients; they produce lactic acid and acetic acid, which lower the pH of the environment and inhibit the colonization of pathogenic microorganisms;

Growing consumer needs have increased the demand for functional food, which means that the food industry introduces more and more interesting and a variety of products. Currently, Europe, Japan, and the United States are the largest markets for functional products [18]. Functional foods must contain one or more compounds that trigger specific changes in the body. In particular, they should help to reduce the risk of civilization diseases, which are the greatest threat to society; for example, cancer, diabetes, heart disease, osteoporosis, neurodegenerative diseases, and hypertension [19]. It is noteworthy that functional foods help to optimize the physiological functions of the body so that it is possible to initiate repair processes and maintain health. It cannot be treated as a drug in specific disease entities, but only as a support in therapy [20]. Compounds that can be used in functional foods

There are different categories of functional foods. The simplest ones are unprocessed conventional foods, for example, tomatoes, kale, raspberries, and broccoli. These foods contain a high content of ellagic acid and lycopene. is the next category is modified foods—this category contains foods that are modified by enrichment with specific ingredients. For example, orange juice with added calcium to support bone health, bread supplemented with folic acid, which is especially dedicated to pregnant women, and margarine enriched with plant stanols. The third category of functional food is a medical food, which is used in specific disease cases and can only be administered under the supervision of a doctor. These foods include supplements for phenylketonuria and diabetes and kidney and liver disease. The latter type is special-purpose food, which includes infant formulas, gluten-free foods, lactose-free foods, and foods used in a slimming diet. Therefore, it may be one of the food products that provides the necessary nutrients. In the case of some categories of food, for example gluten-free food, some components of the material

Fermented foods are grouped as functional foods.  Fermented foods can be obtained by the spontaneous or controlled growth of microorganisms and the enzymatic conversion of their main components. Currently, fermented foods can be produced very fast, which allows for the production of thousands of various products [24]. The fermentation process of some food products gives them new health properties and features that were not present in the starting material. Furthermore, recent clinical trials have shown an existing relationship between the consumption of fermented milk products and maintaining a healthy body weight [25]. Other studies have shown that regular consumption of fermented foods such

*DOI: http://dx.doi.org/10.5772/intechopen.94913*

#### *The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria… DOI: http://dx.doi.org/10.5772/intechopen.94913*

The word "probiotic" was borrowed from Greek, wherein "probios" means "for life." Probiotics are mainly bacterial strains from the genera *Lactobacillus* and *Bifidobacterium*. However, before a strain is considered a probiotic, clinical trials must be conducted to prove its health-promoting properties [15, 16]. There should be mutual benefit between the human body and the probiotic bacteria (on the basis of symbiosis). The intestines are one of the most important organs in maintaining the body's normal immunity. About 70% of the entire population of immune cells is associated with intestinal mucosa [1]. Literature shows that probiotics regulate the functioning of the bacterial microflora in the intestines through certain mechanisms [17]; for example, they compete with the pathogenic bacteria for the same receptors and nutrients; they produce lactic acid and acetic acid, which lower the pH of the environment and inhibit the colonization of pathogenic microorganisms; they produce mucus; and finally, they synthesize B vitamins.

Growing consumer needs have increased the demand for functional food, which means that the food industry introduces more and more interesting and a variety of products. Currently, Europe, Japan, and the United States are the largest markets for functional products [18]. Functional foods must contain one or more compounds that trigger specific changes in the body. In particular, they should help to reduce the risk of civilization diseases, which are the greatest threat to society; for example, cancer, diabetes, heart disease, osteoporosis, neurodegenerative diseases, and hypertension [19]. It is noteworthy that functional foods help to optimize the physiological functions of the body so that it is possible to initiate repair processes and maintain health. It cannot be treated as a drug in specific disease entities, but only as a support in therapy [20]. Compounds that can be used in functional foods are polyphenols, sterols, carotenoids, probiotics, and prebiotics [21].

There are different categories of functional foods. The simplest ones are unprocessed conventional foods, for example, tomatoes, kale, raspberries, and broccoli. These foods contain a high content of ellagic acid and lycopene. is the next category is modified foods—this category contains foods that are modified by enrichment with specific ingredients. For example, orange juice with added calcium to support bone health, bread supplemented with folic acid, which is especially dedicated to pregnant women, and margarine enriched with plant stanols. The third category of functional food is a medical food, which is used in specific disease cases and can only be administered under the supervision of a doctor. These foods include supplements for phenylketonuria and diabetes and kidney and liver disease. The latter type is special-purpose food, which includes infant formulas, gluten-free foods, lactose-free foods, and foods used in a slimming diet. Therefore, it may be one of the food products that provides the necessary nutrients. In the case of some categories of food, for example gluten-free food, some components of the material are removed to avoid the aggravation of the disease [22].

Fermented foods are grouped as functional foods.  Fermented foods can be obtained by the spontaneous or controlled growth of microorganisms and the enzymatic conversion of their main components. Currently, fermented foods can be produced very fast, which allows for the production of thousands of various products [24]. The fermentation process of some food products gives them new health properties and features that were not present in the starting material. Furthermore, recent clinical trials have shown an existing relationship between the consumption of fermented milk products and maintaining a healthy body weight [25]. Other studies have shown that regular consumption of fermented foods such as yogurt reduces the risk of heart disease and type 2 diabetes [26, 27].

*Milk Substitutes - Selected Aspects*

**beverages**

functional foods helps to reduce the risk of developing diseases of affluence, such as diabetes, obesity, or cancer. Buckwheat beverage enriched with LAB and bifidobacterial is one such functional food. Its unique taste and nutritional value might be utilized to develop a new product dedicated to people with disorders of the digestive

Fermented plant-based products represent a better way to substitute dairy products that cannot be consumed by people with food allergies or intolerance. The plant-based products gain a pro-health value after the process of fermentation and at the same time, they require minimal processing. Furthermore, the probiotic LAB have a positive effect on human health by regulating the functions of the intestinal microbiota. They keep the digestive system healthy and increase immunity. They have anticarcinogenic and antiallergenic effects [1]. Food intolerances are not related to the immune system; they are caused by sensitivity to certain food ingredients, e.g. gluten [2]. At present, approximately 20% of the population is affected due to food intolerance [3]. So far, the detailed mechanism of food intolerance is not known, but it may be related to the neuroendocrine system of the digestive system [4]. In the case of treatment available for gluten intolerance, elimination of gluten from the diet is recommended. Any amount of gluten might be harmful to individuals who are gluten-intolerant. According to the literature, more than 50–100 mg of

system, as well as for people who want to stay healthy.

gluten per day can prove to be harmful to such individuals [5].

**2. Plant-based beverages as an alternative for dairy-based probiotic**

Buckwheat is a dicotyledonous plant and is referred to as a pseudocereal. It is classified as a secondary plant. It has a tap root system, which is 1 meter long, and has a straight stem, 60–90 cm high, and brown in color; it bears pink or white flowers. Different products are made from different parts of the plant. The grains are used to produce buckwheat flour and buckwheat, while straw, after threshing the seeds, is added to various types of fodder. During the flowering season, buckwheat provides nectar to the bees [6, 7]. Buckwheat kernels contain glutenfree protein and have well-balanced amino acid profile. The flour is a rich source of minerals such as copper, zinc, manganese, potassium, magnesium, phosphorus, and potassium. It is also rich in polyphenols such as rutin, orientin, vitexin, quercetin, isovitoxin, and isoorientin. Among the aforementioned polyphenols, rutin shows the strongest anti-inflammatory, anticancer, and protective effect. In terms of flavonoid content, tartar buckwheat seeds contain approximately 40 mg/g, whereas common buckwheat seeds contain 10 mg/g [8–10]. A previous study [11] reported that sucrose is the predominant sugar in buckwheat, whereas xylose, glucose, arabinose, and melibiose are present in much smaller quantities. According to a previous study [12], sucrose accumulates in large quantities when the dry matter content is increased. It mainly occurs in the central part of the ovule and the seed coat. Buckwheat also contains *R*-tocopherol, which shows vitamin E activity [13]. In addition, the ethanolic extract of buckwheat contains four catechins: epicatechin, catechin 7-O-β-D-glucopyranoside, (−)-epicatechin 3-O-p-hydroxybenzoate, and (−)-epicatechin-3-O-(3,4-di-O) gallate-methyl [13]. It should be noted that the content of individual components in the plant may change depending on the environmental factors such as temperature, UV radiation, and damage caused by pests. Genetic factors are also of great importance, and the influence of the height of cultivation to sea level has been recently

**78**

demonstrated [14].

## **3. Buckwheat beverages fermented with industrial probiotic cultures**

In this study, we aimed to investigate the effect of four different bacterial cultures containing LAB and bifidobacteria on the selected features of buckwheat beverage. With regard to this, we performed fermentation of the selected cultures with buckwheat beverages and evaluated the parameters.

Fermented plant beverages are very popular in Asia and Africa, for example, boza, togwa, mahewu, makgeolli, or hardalie. The most popular plant-based fermented beverage in Poland and throughout Eastern Europe is kvass. It is a product of milk-alcohol fermentation of wholemeal bread with the addition of yeast, water, and a small amount of sugar. The microorganisms present in kvass are *Lactobacillus casei*, *Leuconostoc mesenteroides*, and *Saccharomyces cerevisiae* [28].

In recent years, many studies have reported the properties of plant-based fermented beverages. The most important feature of this type of product is the ability of LAB to carry out effective fermentation, and the pH value of the resulting product is an important parameter, which indicates the effectiveness of the fermentation process. In this study, this parameter was checked both during the fermentation process and after its completion (28 days).

Kowalska [29] used four yogurt starter cultures to ferment the buckwheat beverage: YO-MIX 207, YO-MIX 205, ABY-3, and VEGE 033. The microbial composition of the starters was as follows:

Buckwheat beverage was prepared with 200 g of boiled buckwheat and blended with 3000 mL of drinking water. Prior to the process of sterilization, the beverage was strained through a fine sieve to get rid of the groats. The strained beverage was sterilized at 121°C for 15 min [30]. Based on the recipe of the buckwheat beverage, which was obtained by mixing the buckwheat in water in the proportion 1:15, the nutritional value of 100 g of buckwheat beverage was calculated [29]:

The average water content of buckwheat beverage was 87.9% [30].

Kowalska [29] reported that the initial pH of buckwheat (beverage before fermentation at 37°C for 5 h) was on an average 6.550 for the samples intended for fermentation with YO-MIX 207, YO-MIX 205, and ABY-3 cultures, and 6.400 for the samples

**81**

**Table 1.**

*(based on [29]).*

*The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria…*

intended for fermentation with VEGE 033 culture (**Table 1**). The most effective fermentation process was observed in the case of beverage fermented with YO-MIX 207 culture, followed by the beverage fermented with YO-MIX 205. Within 1–2 h of fermentation, both beverages reached an average pH value of 4.8, which was statistically significantly from that of before fermentation (**Table 1**). ABY3 and VEGE 033 cultures were less efficient in terms of acidification, in which case, the pH value did not increase until 3–4 h of the fermentation process. After fermentation for 5 h, all of the beverages reached a pH of 4.5–4.9, which means that all the bacteria carried out the fermentation process efficiently [29]. A previous study [31] also reported similar results for soybean beverage fermented with *S. thermophilus*. However, a previous study [32] conducted on barley malt fermented with *Lactobacillus plantarum* (NCIMB 8826) and *L. acidophilus* (NCIMB 8821) recorded a pH value of approximately 4.0. This difference in pH value might be because of the specificity of plant matrices, as

well as the use of various bacterial cultures for fermentation (**Table 2**).

variation in pH value during refrigerated storage.

genus *Bifidobacterium* [35].

**Fermentation time [h]**

was not observed in the research conducted by Kowalska [29].

Kowalska [29] also measured the pH of buckwheat beverage during 28 days of refrigerated storage. During refrigerated storage, the most stable pH value was recorded for buckwheat beverage fermented with VEGE 033, ABY-3, and YO-MIX 205 cultures. However, the beverage fermented with YO-MIX 207 culture showed

**Table 1** shows the pH value of buckwheat beverage before and after fermentation with cultures tested by Kowalska [29]. Similar results were obtained by Ziarno et al. [33]. They reported the change in pH value of bean beverage (initial pH of 6.58) after fermentation, which was 4.47 and 4.45 when fermented with YO-MIX 205 and ABY-3 cultures, respectively. At 6°C, the pH value of beverages fermented with YO-MIX 205 and ABY-3 cultures respectively decreased to 4.40 and 4.39 on day 7, 4.34 and 4.29 on day 21, and 4.33 and 4.27 on day 28 [33]. This shows that LAB continued the process of fermentation during the entire storage period, which

Bacterial cell count is a very important parameter in determining the quality of the product and its health properties [34]. Manufacturers frequently check this parameter in fermented beverages. The minimum acceptable number of live LAB cells that should be present in fermented beverages is 7 log(CFU/mL) and at least 6 log(CFU/mL) for strains with probiotic properties, including probiotics of the

Kowalska [29] found that the changes in the number of live LAB and bifidobacteria in beverages fermented with the YO-MIX 205 and YO-MIX 207 cultures were

 6.550 ± 0.212a 6.550 ± 0.212a 6.550 ± 0.212a 6.400 ± 0.000a 5.185 ± 0.481b 5.020 ± 0.389b 5.610 ± 0.721ab 5.770 ± 0.000ab 4.840 ± 0.226b 4.805 ± 0.163b 5.085 ± 0.262b 5.170 ± 0.000b 4.730 ± 0.127<sup>b</sup> 4.675 ± 0.163b 4.860 ± 0.085b 4.910 ± 0.000b 4.640 ± 0.057<sup>b</sup> 4.600 ± 0.212b 4.825 ± 0.106b 4.880 ± 0.000b 4.590 ± 0.127<sup>b</sup> 4.595 ± 0.276b 4.790 ± 0.156b 4.950 ± 0.000b

*Note: a,b—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

*pH values of buckwheat beverage during the fermentation process (mean ± standard deviation)*

**Buckwheat beverages fermented by: YO-MIX 207 YO-MIX 205 ABY-3 VEGE 033**

*DOI: http://dx.doi.org/10.5772/intechopen.94913*

#### *The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria… DOI: http://dx.doi.org/10.5772/intechopen.94913*

intended for fermentation with VEGE 033 culture (**Table 1**). The most effective fermentation process was observed in the case of beverage fermented with YO-MIX 207 culture, followed by the beverage fermented with YO-MIX 205. Within 1–2 h of fermentation, both beverages reached an average pH value of 4.8, which was statistically significantly from that of before fermentation (**Table 1**). ABY3 and VEGE 033 cultures were less efficient in terms of acidification, in which case, the pH value did not increase until 3–4 h of the fermentation process. After fermentation for 5 h, all of the beverages reached a pH of 4.5–4.9, which means that all the bacteria carried out the fermentation process efficiently [29]. A previous study [31] also reported similar results for soybean beverage fermented with *S. thermophilus*. However, a previous study [32] conducted on barley malt fermented with *Lactobacillus plantarum* (NCIMB 8826) and *L. acidophilus* (NCIMB 8821) recorded a pH value of approximately 4.0. This difference in pH value might be because of the specificity of plant matrices, as well as the use of various bacterial cultures for fermentation (**Table 2**).

Kowalska [29] also measured the pH of buckwheat beverage during 28 days of refrigerated storage. During refrigerated storage, the most stable pH value was recorded for buckwheat beverage fermented with VEGE 033, ABY-3, and YO-MIX 205 cultures. However, the beverage fermented with YO-MIX 207 culture showed variation in pH value during refrigerated storage.

**Table 1** shows the pH value of buckwheat beverage before and after fermentation with cultures tested by Kowalska [29]. Similar results were obtained by Ziarno et al. [33]. They reported the change in pH value of bean beverage (initial pH of 6.58) after fermentation, which was 4.47 and 4.45 when fermented with YO-MIX 205 and ABY-3 cultures, respectively. At 6°C, the pH value of beverages fermented with YO-MIX 205 and ABY-3 cultures respectively decreased to 4.40 and 4.39 on day 7, 4.34 and 4.29 on day 21, and 4.33 and 4.27 on day 28 [33]. This shows that LAB continued the process of fermentation during the entire storage period, which was not observed in the research conducted by Kowalska [29].

Bacterial cell count is a very important parameter in determining the quality of the product and its health properties [34]. Manufacturers frequently check this parameter in fermented beverages. The minimum acceptable number of live LAB cells that should be present in fermented beverages is 7 log(CFU/mL) and at least 6 log(CFU/mL) for strains with probiotic properties, including probiotics of the genus *Bifidobacterium* [35].

Kowalska [29] found that the changes in the number of live LAB and bifidobacteria in beverages fermented with the YO-MIX 205 and YO-MIX 207 cultures were

#### *Note: a,b—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

#### **Table 1.**

*Milk Substitutes - Selected Aspects*

**3. Buckwheat beverages fermented with industrial probiotic cultures**

In this study, we aimed to investigate the effect of four different bacterial cultures containing LAB and bifidobacteria on the selected features of buckwheat beverage. With regard to this, we performed fermentation of the selected cultures

Fermented plant beverages are very popular in Asia and Africa, for example, boza, togwa, mahewu, makgeolli, or hardalie. The most popular plant-based fermented beverage in Poland and throughout Eastern Europe is kvass. It is a product of milk-alcohol fermentation of wholemeal bread with the addition of yeast, water, and a small amount of sugar. The microorganisms present in kvass are *Lactobacillus*

In recent years, many studies have reported the properties of plant-based fermented beverages. The most important feature of this type of product is the ability of LAB to carry out effective fermentation, and the pH value of the resulting product is an important parameter, which indicates the effectiveness of the fermentation process. In this study, this parameter was checked both during the fermenta-

Kowalska [29] used four yogurt starter cultures to ferment the buckwheat beverage: YO-MIX 207, YO-MIX 205, ABY-3, and VEGE 033. The microbial composition

a.ABY-3 (Chr. Hansen, Denmark)—Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus acidophilus La-5, Bifidobacterium

b.YO-MIX 207 (DuPont Danisco, Denmark)—*S. thermophilus*, L. delbrueckii

d.VEGE 033 LYO (DuPont Danisco, Denmark)—*S. thermophilus*, L. delbrueckii

Buckwheat beverage was prepared with 200 g of boiled buckwheat and blended with 3000 mL of drinking water. Prior to the process of sterilization, the beverage was strained through a fine sieve to get rid of the groats. The strained beverage was sterilized at 121°C for 15 min [30]. Based on the recipe of the buckwheat beverage, which was obtained by mixing the buckwheat in water in the proportion 1:15, the

c.YO-MIX 205 LYO (DuPont Danisco, Denmark)—*S. thermophilus*, L.

subsp. bulgaricus, L. acidophilus, Bifidobacterium lactis,

delbrueckii subsp. bulgaricus, L. acidophilus, B. lactis,

subsp. bulgaricus, L. acidophilus NCFM, B. lactis HN019.

nutritional value of 100 g of buckwheat beverage was calculated [29]:

The average water content of buckwheat beverage was 87.9% [30].

Kowalska [29] reported that the initial pH of buckwheat (beverage before fermentation at 37°C for 5 h) was on an average 6.550 for the samples intended for fermentation with YO-MIX 207, YO-MIX 205, and ABY-3 cultures, and 6.400 for the samples

• Fat—0.16 g (including 0.04 g of saturated acids)

• Carbohydrates—4.69 g (including 0.16 g of sugars)

with buckwheat beverages and evaluated the parameters.

tion process and after its completion (28 days).

of the starters was as follows:

animalis subsp. lactis BB-12,

*casei*, *Leuconostoc mesenteroides*, and *Saccharomyces cerevisiae* [28].

**80**

• Proteins—0.75 g.

*pH values of buckwheat beverage during the fermentation process (mean ± standard deviation) (based on [29]).*

#### **Table 2.**

*pH values of buckwheat beverage fermented with different starter cultures (mean values and standard deviations) (based on [29]).*

very similar. Interestingly, after fermentation, there was a slight reduction in the number of viable bacterial cells compared to the state before fermentation. In addition, during the refrigerated storage of the fermented beverage, there were fluctuations in the number of LAB cells, both lactobacilli and lactic streptococci, as well as bifidobacteria. The number of viable cells of lactobacilli, lactic streptococci, and bifidobacteria on day 28 was over 7 log(CFU/mL), which indicated the potential health-promoting properties of the tested beverages fermented with the YO-MIX 207 and YO-MIX 205 cultures.

The smallest variation in the population of lactobacilli, lactic streptococci, and bifidobacteria was recorded for beverages fermented with ABY-3 culture (**Table 3**) [29]. Contrary to buckwheat beverages fermented with the YO-MIX 207 (**Table 4**) and YO-MIX 205 (**Table 5**), there were no such significant changes in the number of bacterial cells. After fermentation, the number of bifidobacterial cells decreased the most. After 7 days of refrigerated storage (4°C), there was a slight change in the number of lactobacilli, lactic streptococci, and bifidobacteria. After 28 days of storage, the average bacterial cell count was 8.0 log(CFU/mL) for lactobacilli, 7.8 log(CFU/mL) for lactic streptococci, and 8.0 log(CFU/mL) for bifidobacteria. The number of viable cells of lactobacilli, lactic streptococci, and bifidobacteria on day 28 was over 7 log(CFU/mL), which indicated the potentially healthpromoting properties of the tested buckwheat beverages fermented with the ABY-3 culture [29].

According to Kowalska [29], in the case of buckwheat beverage fermented with VEGE 033 (**Table 6**), the greatest proportion in the population of bacterial cells prior to fermentation were lactic streptococci [29]. In the beverages fermented with the VEGE 033 culture, the lower number of bifidobacterial cells was found (during the entire period of cooling storage) compared to the buckwheat beverage fermented with ABY-3 culture. On the 7th day of storage of the samples of buckwheat beverages fermented with the VEGE 033 culture, the number of streptococcal cells was on average 8.2 log(CFU/mL). The number of viable lactobacilli, lactic streptococci, and bifidobacteria cells in the beverage fermented with VEGE 033 culture on day 28 was over 7 log(CFU/mL) [29].

A previous study conducted on rice beverage reported low counts of bacterial cells [36]. Prior to fermentation, the number of bacterial cells in rice beverage was lower than that observed for buckwheat beverage in the research conducted by Kowalska [29] - the population of LAB was 5.0 log(CFU/mL). However, after 16-hour fermentation process, the bacterial population increased to 8.1 log(CFU/mL) and remained at this level until the end of the fermentation process [29]. However, the previous study [37] reported that after fermentation of corn or rice-based beverages, the microbial cell population was at the level of 7–8 log(CFU/mL). This number indicates that the product has probiotic properties [38].

**83**

but their strain was different.

*The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria…*

After fermentation 7.9 ± 0.3a 7.7 ± 0.1b 8.0 ± 0.3a 7 day of storage 8.1 ± 0.3a 8.2 ± 0.2ab 8.0 ± 0.2a 28 days of storage 8.0 ± 0.2a 8.0 ± 0.2ab 7.8 ± 0.1a

*The population of live cells of lactic acid bacteria and bifidobacteria in buckwheat beverage fermented with ABY-3 culture and stored for 28 days under refrigerated condition (mean ± standard deviation) (based on [29]).*

After fermentation 7.8 ± 0.2b 7.2 ± 0.1c 7.9 ± 0.3a 7 day of storage 8.0 ± 0.3b 8.1 ± 0.3ab 8.0 ± 0.4a 28 days of storage 7.8 ± 0.0b 7.7 ± 0.1b 7.7 ± 0.3a

*The population of live cells of lactic acid bacteria and bifidobacteria in buckwheat beverage fermented with YO-MIX 207 culture and stored for 28 days under refrigerated condition (mean ± standard deviation)*

After fermentation 7.6 ± 0.2a 7.0 ± 0.2a 7.8 ± 0.5a 7 day of storage 7.9 ± 0.6a 7.6 ± 0.7a 8.1 ± 0.5a 28 days of storage 7.6 ± 0.1a 7.5 ± 0.3a 7.8 ± 0.2a

*—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

*Note: a–c—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

*Note: a,b—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

**Number of bifidobacteria [log(CFU/mL)]**

8.2 ± 0.2a 8.2 ± 0.4a 8.2 ± 0.4a

**Number of bifidobacteria [log(CFU/mL)]**

8.7 ± 0.3a 8.5 ± 0.2a 8.4 ± 1.1a

**Number of bifidobacteria [log(CFU/mL)]**

7.9 ± 0.8a 7.8 ± 0.7a 8.4 ± 0.8a

**Number of lactic streptococci [log(CFU/mL)]**

**Number of lactic streptococci [log(CFU/mL)]**

**Number of lactic streptococci [log(CFU/mL)]**

**Number of lactobacilli [log(CFU/mL)]**

**Number of lactobacilli [log(CFU/mL)]**

**Number of lactobacilli [log(CFU/mL)]**

However, another group of researchers [39] used different strains of LAB for fermentation of soy milk, including *L. delbrueckii* subsp. bulgaricus and *L. acidophilus*, which were also used in this study. The cell population of all cultures was 8 log(CFU/mL), which is similar to the results of this study with buckwheat. In each bacterial culture, *L. delbrueckii* subsp. bulgaricus and *L. acidophilus* were present,

*The population of live cells of lactic acid bacteria and bifidobacteria in buckwheat beverage fermented with YO-MIX 205 culture and stored for 28 days under refrigerated conditions (mean ± standard deviation)*

Kowalska [29] found that in all plant-based products, there were similarities in the population of LAB, despite the diversity of the *Lactobacillus* strains used. The

*DOI: http://dx.doi.org/10.5772/intechopen.94913*

**Determination**

**Determination**

**time**

Before fermentation

**Table 3.**

**time**

Before fermentation

**Table 4.**

*(based on [29]).*

**time**

Before fermentation

*Note: a*

**Table 5.**

*(based on [29]).*

**Determination**

*The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria… DOI: http://dx.doi.org/10.5772/intechopen.94913*

#### **Table 3.**

*Milk Substitutes - Selected Aspects*

**Storage time [day]**

**Table 2.**

*deviations) (based on [29]).*

207 and YO-MIX 205 cultures.

day 28 was over 7 log(CFU/mL) [29].

indicates that the product has probiotic properties [38].

culture [29].

very similar. Interestingly, after fermentation, there was a slight reduction in the number of viable bacterial cells compared to the state before fermentation. In addition, during the refrigerated storage of the fermented beverage, there were fluctuations in the number of LAB cells, both lactobacilli and lactic streptococci, as well as bifidobacteria. The number of viable cells of lactobacilli, lactic streptococci, and bifidobacteria on day 28 was over 7 log(CFU/mL), which indicated the potential health-promoting properties of the tested beverages fermented with the YO-MIX

*pH values of buckwheat beverage fermented with different starter cultures (mean values and standard*

*Note: a,b—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

 4.590 ± 0.127<sup>b</sup> 4.595 ± 0.276a 4.790 ± 0.156a 4.950 ± 0.000a 4.750 ± 0.071ab 4.750 ± 0.071a 4.850 ± 0.071a 4.900 ± 0.000a 4.875 ± 0.035ab 4.850 ± 0.071a 4.850 ± 0.071a 5.000 ± 0.000a 4.920 ± 0.028a 4.935 ± 0.049a 4.925 ± 0.035a 5.000 ± 0.000a

**Buckwheat beverage fermented by: YO-MIX 207 YO-MIX 207 YO-MIX 207 YO-MIX 207**

The smallest variation in the population of lactobacilli, lactic streptococci, and bifidobacteria was recorded for beverages fermented with ABY-3 culture (**Table 3**) [29]. Contrary to buckwheat beverages fermented with the YO-MIX 207 (**Table 4**) and YO-MIX 205 (**Table 5**), there were no such significant changes in the number of bacterial cells. After fermentation, the number of bifidobacterial cells decreased the most. After 7 days of refrigerated storage (4°C), there was a slight change in the number of lactobacilli, lactic streptococci, and bifidobacteria. After 28 days of storage, the average bacterial cell count was 8.0 log(CFU/mL) for lactobacilli, 7.8 log(CFU/mL) for lactic streptococci, and 8.0 log(CFU/mL) for bifidobacteria. The number of viable cells of lactobacilli, lactic streptococci, and bifidobacteria on day 28 was over 7 log(CFU/mL), which indicated the potentially health-

promoting properties of the tested buckwheat beverages fermented with the ABY-3

A previous study conducted on rice beverage reported low counts of bacterial cells [36]. Prior to fermentation, the number of bacterial cells in rice beverage was lower than that observed for buckwheat beverage in the research conducted by Kowalska [29] - the population of LAB was 5.0 log(CFU/mL). However, after 16-hour fermentation process, the bacterial population increased to 8.1 log(CFU/mL) and remained at this level until the end of the fermentation process [29]. However, the previous study [37] reported that after fermentation of corn or rice-based beverages, the microbial cell population was at the level of 7–8 log(CFU/mL). This number

According to Kowalska [29], in the case of buckwheat beverage fermented with VEGE 033 (**Table 6**), the greatest proportion in the population of bacterial cells prior to fermentation were lactic streptococci [29]. In the beverages fermented with the VEGE 033 culture, the lower number of bifidobacterial cells was found (during the entire period of cooling storage) compared to the buckwheat beverage fermented with ABY-3 culture. On the 7th day of storage of the samples of buckwheat beverages fermented with the VEGE 033 culture, the number of streptococcal cells was on average 8.2 log(CFU/mL). The number of viable lactobacilli, lactic streptococci, and bifidobacteria cells in the beverage fermented with VEGE 033 culture on

**82**

*The population of live cells of lactic acid bacteria and bifidobacteria in buckwheat beverage fermented with ABY-3 culture and stored for 28 days under refrigerated condition (mean ± standard deviation) (based on [29]).*

#### **Table 4.**

*The population of live cells of lactic acid bacteria and bifidobacteria in buckwheat beverage fermented with YO-MIX 207 culture and stored for 28 days under refrigerated condition (mean ± standard deviation) (based on [29]).*

#### **Table 5.**

*The population of live cells of lactic acid bacteria and bifidobacteria in buckwheat beverage fermented with YO-MIX 205 culture and stored for 28 days under refrigerated conditions (mean ± standard deviation) (based on [29]).*

However, another group of researchers [39] used different strains of LAB for fermentation of soy milk, including *L. delbrueckii* subsp. bulgaricus and *L. acidophilus*, which were also used in this study. The cell population of all cultures was 8 log(CFU/mL), which is similar to the results of this study with buckwheat. In each bacterial culture, *L. delbrueckii* subsp. bulgaricus and *L. acidophilus* were present, but their strain was different.

Kowalska [29] found that in all plant-based products, there were similarities in the population of LAB, despite the diversity of the *Lactobacillus* strains used. The

#### **Table 6.**

*The population of live cells of lactic acid bacteria and bifidobacteria in buckwheat beverage fermented with VEGE 033 culture and stored for 28 days under refrigerated condition (mean ± standard deviation) (based on [29]).*

good growth of LAB in plant-based beverages can be explained by the high amounts of mono and disaccharides in the plant media.

A previous study performed fermentation of bean beverages with ABY-3 culture [33]. Prior to fermentation, the number of viable lactobacilli was 7.7 log(CFU/mL), which gradually decreased during the cold storage. On days 7 and 28 of storage, the population of lactobacilli was 7.5 log(CFU/mL) and 6.9 log(CFU/mL), respectively. According to a previous study [30], the observed lower bacterial cells after the cold storage period may result from antimicrobial compounds produced by bacteria, e.g. hydrogen peroxide, bacteriocins, or organic acids. In contrast, in the research conducted by Kowalska [29], the number of viable lactobacilli in the buckwheat beverage fermented with ABY-3 culture was slightly higher. Prior to fermentation, on days 7 and 28 of storage, the number of viable lactobacilli was 8.2 log(CFU/mL) and 8.0 log(CFU/mL), respectively. The better growth on buckwheat substrate might be due to higher sugar content and availability in plant media.

Kowalska [29] verified the content of carbohydrates using high-performance liquid chromatography. The results showed the presence of 7 carbohydrates: xylose, melibiose, fructose, arabinose, glucose, sucrose, and maltose. The initial (before fermentation) content of carbohydrate in the fermented buckwheat beverage was 4.598 g in 100 g of the product. The chromatographic analysis includes only a few selected carbohydrates, whereas the calculated value of carbohydrate content takes into account all such compounds, including starch. Therefore, it can be concluded that as a result of the cooking and sterilization of buckwheat beverage in an aqueous solution, some complex carbohydrates or polysaccharides might be released, which were determined by chromatography [29].

Immediately after the end of fermentation of buckwheat beverages, the highest total carbohydrate content was found in the beverage fermented with the ABY-3 culture (**Table 7**), whereas the lowest was found in the beverage fermented with the YO-MIX 207 culture (**Table 8**). It should be noted that both the ABY-3 culture and the YO-MIX 207 culture had a rich microbiological composition, which not only included LAB but also included bifidobacteria of different strains [29].

In the case of beverage fermented with YO-MIX 205 culture (**Table 9**), we obtained statistically significant differences in terms of carbohydrate content before and after fermentation and during cold storage.

Contrary to the buckwheat beverages fermented with the YO-MIX 205 and YO-MIX 207 cultures, the beverage fermented with VEGE 033 culture (**Table 10**) contained a low amount of xylose after fermentation. In this case, the xylose content decreased slightly. As in the beverages fermented with the

**85**

*The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria…*

Xylose 0.000e 0.129e 0.065f 0.193b Fructose 0.096d 0.322b 0.132e 0.000e Arabinose 0.000e 0.241d 0.294c 0.000e Glucose 2.958a 0.280c 0.251d 0.152c Melibiose 0.000e 0.000f 0.318b 0.153c Sucrose 1.544b 2.300a 1.591a 0.698a Maltose 0.218c 0.000f 0.000g 0.000e All 4.598 3.273 2.650 1.196

**After fermentation**

**After fermentation**

Xylose 0.000f 0.000f 0.000e 0.143c Fructose 0.096d 0.115c 0.076d 0.164b Arabinose 0.000e 0.069d 0.086c 0.000g Glucose 2.958a 0.204b 0.186b 0.102d Melibiose 0.000e 0.000e 0.000f 0.087e Sucrose 1.544b 1.436a 1.388a 0.751a Maltose 0.218c 0.000f 0.000f 0.000f All 4.598 1.824 1.736 1.247

**After 7 days of storage**

**After 7 days of storage**

**After 28 days of storage**

**After 28 days of storage**

**Before fermentation**

*Note: a–e—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

*Content of carbohydrates in buckwheat beverages fermented with ABY-3 culture (based on [29]).*

**Before fermentation**

YO-MIX 205 and YO-MIX 207 cultures, the content of sucrose, glucose, and maltose also decreased, and the content of arabinose increased. The chromatographic analysis also did not detect the presence of melibiose. Statistical analysis showed significant differences in the results of carbohydrate content

*Content of carbohydrates in buckwheat beverages fermented with YO-MIX 207 culture (based on [29]).*

*Note: a–g—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

Our results show differences in the fermentation abilities of the tested starter cultures, resulting from different biochemical activities (mainly saccharolytic and

It can be assumed that the changes in the content of carbohydrates during refrigerated storage were due to the changes taking place in the analyzed samples; for example, the biochemical activity of LAB and bifidobacteria, as well as enzymatic changes [29]. Due to the lack of information, it is difficult to compare the results of

A previous study [40] reported contradictory results with respect to sugar content in the cooked buckwheat wort. According to the result of the aforementioned study, glucose was present in the highest quantities. However, in this study, sucrose was found to be the highest after fermentation and after the storage period, which

fermentation) of the strains present in the tested cultures [29].

was most likely the result of starch decomposition.

during cold storage of the samples.

this study with that of others.

*DOI: http://dx.doi.org/10.5772/intechopen.94913*

**Carbohydrates [g/ 100 g beverage]**

**Table 7.**

**Table 8.**

**Carbohydrates [g/ 100 g beverage]** *The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria… DOI: http://dx.doi.org/10.5772/intechopen.94913*

**Table 7.**

*Milk Substitutes - Selected Aspects*

**Determination**

**time**

Before fermentation

**Table 6.**

*(based on [29]).*

of mono and disaccharides in the plant media.

**Number of lactobacilli [log(CFU/mL)]**

were determined by chromatography [29].

and after fermentation and during cold storage.

good growth of LAB in plant-based beverages can be explained by the high amounts

*The population of live cells of lactic acid bacteria and bifidobacteria in buckwheat beverage fermented with VEGE 033 culture and stored for 28 days under refrigerated condition (mean ± standard deviation)*

After fermentation 7.1 ± 0.1d 7.3 ± 0.0b 7.9 ± 0.0c 7 day of storage 8.1 ± 0.1b 8.2 ± 0.2a 8.2 ± 0.1b 28 days of storage 7.7 ± 0.0c 7.4 ± 0.1b 7.5 ± 0.0d

*Note: a–d—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

Kowalska [29] verified the content of carbohydrates using high-performance liquid chromatography. The results showed the presence of 7 carbohydrates: xylose, melibiose, fructose, arabinose, glucose, sucrose, and maltose. The initial (before fermentation) content of carbohydrate in the fermented buckwheat beverage was 4.598 g in 100 g of the product. The chromatographic analysis includes only a few selected carbohydrates, whereas the calculated value of carbohydrate content takes into account all such compounds, including starch. Therefore, it can be concluded that as a result of the cooking and sterilization of buckwheat beverage in an aqueous solution, some complex carbohydrates or polysaccharides might be released, which

Immediately after the end of fermentation of buckwheat beverages, the highest total carbohydrate content was found in the beverage fermented with the ABY-3 culture (**Table 7**), whereas the lowest was found in the beverage fermented with the YO-MIX 207 culture (**Table 8**). It should be noted that both the ABY-3 culture and the YO-MIX 207 culture had a rich microbiological composition, which not only

might be due to higher sugar content and availability in plant media.

included LAB but also included bifidobacteria of different strains [29].

In the case of beverage fermented with YO-MIX 205 culture (**Table 9**), we obtained statistically significant differences in terms of carbohydrate content before

Contrary to the buckwheat beverages fermented with the YO-MIX 205 and YO-MIX 207 cultures, the beverage fermented with VEGE 033 culture (**Table 10**) contained a low amount of xylose after fermentation. In this case, the xylose content decreased slightly. As in the beverages fermented with the

A previous study performed fermentation of bean beverages with ABY-3 culture [33]. Prior to fermentation, the number of viable lactobacilli was 7.7 log(CFU/mL), which gradually decreased during the cold storage. On days 7 and 28 of storage, the population of lactobacilli was 7.5 log(CFU/mL) and 6.9 log(CFU/mL), respectively. According to a previous study [30], the observed lower bacterial cells after the cold storage period may result from antimicrobial compounds produced by bacteria, e.g. hydrogen peroxide, bacteriocins, or organic acids. In contrast, in the research conducted by Kowalska [29], the number of viable lactobacilli in the buckwheat beverage fermented with ABY-3 culture was slightly higher. Prior to fermentation, on days 7 and 28 of storage, the number of viable lactobacilli was 8.2 log(CFU/mL) and 8.0 log(CFU/mL), respectively. The better growth on buckwheat substrate

**Number of bifidobacteria [log(CFU/mL)]**

8.7 ± 0.0a 7.1 ± 0.1b 9.0 ± 0.1a

**Number of lactic streptococci [log(CFU/mL)]**

**84**

*Content of carbohydrates in buckwheat beverages fermented with ABY-3 culture (based on [29]).*

#### **Table 8.**

*Content of carbohydrates in buckwheat beverages fermented with YO-MIX 207 culture (based on [29]).*

YO-MIX 205 and YO-MIX 207 cultures, the content of sucrose, glucose, and maltose also decreased, and the content of arabinose increased. The chromatographic analysis also did not detect the presence of melibiose. Statistical analysis showed significant differences in the results of carbohydrate content during cold storage of the samples.

Our results show differences in the fermentation abilities of the tested starter cultures, resulting from different biochemical activities (mainly saccharolytic and fermentation) of the strains present in the tested cultures [29].

It can be assumed that the changes in the content of carbohydrates during refrigerated storage were due to the changes taking place in the analyzed samples; for example, the biochemical activity of LAB and bifidobacteria, as well as enzymatic changes [29]. Due to the lack of information, it is difficult to compare the results of this study with that of others.

A previous study [40] reported contradictory results with respect to sugar content in the cooked buckwheat wort. According to the result of the aforementioned study, glucose was present in the highest quantities. However, in this study, sucrose was found to be the highest after fermentation and after the storage period, which was most likely the result of starch decomposition.

*Note: a–f—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

#### **Table 9.**

*Content of carbohydrates in buckwheat beverages fermented with YO-MIX 205 culture (based on [29]).*

#### **Table 10.**

*Content of carbohydrates in buckwheat beverages fermented by VEGE 033 culture (based on [29]).*

A previous study [41] reported that sucrose was the predominant carbohydrate, whereas xylose, glucose, arabinose, and melibiose were present in much smaller quantities. Another study [42] reported that with an increasing amount of water and lengthening heating time, the content of glucose increases.

According to the literature [43], fermentation of buckwheat beverages with the use of *Propionibacterium freudenreichii* subsp. *shermanii* resulted in a significant increase in the content of fructose, glucose, and galactose. In addition, there was a significant increase in sucrose content.

### **4. Conclusion**

The results of this study indicate a high potential of fermented buckwheat beverage as a probiotic product with pro-health properties. The demand for gluten-free cereal beverages is growing among people suffering from celiac disease and food intolerance. Good bacterial survival during the storage period allows achieving a therapeutic effect similar to that caused by consuming fermented milk products, such as kefir, buttermilk, or yoghurt. In addition, an additional advantage of the product is the lack of allergenic milk proteins. More and more people are

**87**

**Author details**

**Acknowledgements**

**Conflict of interest**

WULS-SGGW.

this research.

Ewa Kowalska1

(WULS-SGGW), Warsaw, Poland

(WULS-SGGW), Warsaw, Poland

provided the original work is properly cited.

\* and Małgorzata Ziarno2

civilization diseases such as diabetes, obesity, or cancer.

\*Address all correspondence to<EMAIL_ADDRESS>
1 Institute of Horticultural Sciences, Warsaw University of Life Sciences - SGGW

2 Division of Milk Technology, Department of Food Technology and Assessment,

© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium,

Institute of Food Science, Warsaw University of Life Sciences - SGGW

*The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria…*

experiencing side effects after drinking milk and other dairy products such as gas, indigestion, and diarrhea, which are causing them to be excluded from their diet. In such a case, dietary supplements containing probiotic strains are often used to supplement the intestinal microflora and increase the body's immunity. Fermented buckwheat beverages can replace these types of supplements and provide other essential nutrients for the body. The product is dedicated not only to people suffering from disorders of the digestive system but also to healthy people who care about a balanced diet and want to have a healthy lifestyle. In addition to LAB and bifidobacteria, the base of the buckwheat beverage is important, as it is also a medium necessary for the growth of the bacterial population used for fermentation. Our results show that buckwheat can be successfully fermented by LAB and bifidobacteria. Its proven health properties mean that the beverage can be used to prevent

This work was supported by a grant from Warsaw University of Life Sciences -

Authors have declared that they do not have any conflict of interest in publishing

*DOI: http://dx.doi.org/10.5772/intechopen.94913*

*The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria… DOI: http://dx.doi.org/10.5772/intechopen.94913*

experiencing side effects after drinking milk and other dairy products such as gas, indigestion, and diarrhea, which are causing them to be excluded from their diet. In such a case, dietary supplements containing probiotic strains are often used to supplement the intestinal microflora and increase the body's immunity. Fermented buckwheat beverages can replace these types of supplements and provide other essential nutrients for the body. The product is dedicated not only to people suffering from disorders of the digestive system but also to healthy people who care about a balanced diet and want to have a healthy lifestyle. In addition to LAB and bifidobacteria, the base of the buckwheat beverage is important, as it is also a medium necessary for the growth of the bacterial population used for fermentation. Our results show that buckwheat can be successfully fermented by LAB and bifidobacteria. Its proven health properties mean that the beverage can be used to prevent civilization diseases such as diabetes, obesity, or cancer.

## **Acknowledgements**

*Milk Substitutes - Selected Aspects*

**Before fermentation**

*Note: a–f—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

**Before fermentation**

*Content of carbohydrates in buckwheat beverages fermented with YO-MIX 205 culture (based on [29]).*

Xylose 0.000e 0.080e 0.625c 0.705a Fructose 0.096d 0.094d 0.000e 0.000e Arabinose 0.000e 0.741b 1.299b 0.264c Glucose 2.958a 0.237c 0.328d 0.106d Melibiose 0.000e 0.000f 0.000e 0.000e Sucrose 1.544b 1.237a 1.598a 0.338b Maltose 0.218c 0.000f 0.000e 0.000e All 4.598 2.389 3.851 1.413

**After fermentation**

**After fermentation**

Xylose 0.000e 0.000e 0.042e 0.233c Fructose 0.096d 0.099d 0.093d 0.244b Arabinose 0.000e 0.152c 0.873b 0.000f Glucose 2.958a 0.000e 0.000f 0.123d Melibiose 0.000e 0.286b 0.188c 0.075e Sucrose 1.544b 1.514a 1.763a 0.595a Maltose 0.218c 0.000e 0.000f 0.000f All 4.598 2.051 2.959 1.270

**After 7 days of storage**

**After 7 days of storage**

**After 28 days of storage**

**After 28 days of storage**

**Carbohydrates [g/ 100 g beverage]**

**Table 9.**

**Table 10.**

**Carbohydrates [g/ 100 g beverage]**

A previous study [41] reported that sucrose was the predominant carbohydrate, whereas xylose, glucose, arabinose, and melibiose were present in much smaller quantities. Another study [42] reported that with an increasing amount of water

According to the literature [43], fermentation of buckwheat beverages with the use of *Propionibacterium freudenreichii* subsp. *shermanii* resulted in a significant increase in the content of fructose, glucose, and galactose. In addition, there was a

The results of this study indicate a high potential of fermented buckwheat beverage as a probiotic product with pro-health properties. The demand for gluten-free cereal beverages is growing among people suffering from celiac disease and food intolerance. Good bacterial survival during the storage period allows achieving a therapeutic effect similar to that caused by consuming fermented milk products, such as kefir, buttermilk, or yoghurt. In addition, an additional advantage of the product is the lack of allergenic milk proteins. More and more people are

and lengthening heating time, the content of glucose increases.

*Note: a–f—values in columns with the same letter do not differ statistically significantly for* α *= 0.05.*

*Content of carbohydrates in buckwheat beverages fermented by VEGE 033 culture (based on [29]).*

significant increase in sucrose content.

**86**

**4. Conclusion**

This work was supported by a grant from Warsaw University of Life Sciences - WULS-SGGW.

## **Conflict of interest**

Authors have declared that they do not have any conflict of interest in publishing this research.

## **Author details**

Ewa Kowalska1 \* and Małgorzata Ziarno2

1 Institute of Horticultural Sciences, Warsaw University of Life Sciences - SGGW (WULS-SGGW), Warsaw, Poland

2 Division of Milk Technology, Department of Food Technology and Assessment, Institute of Food Science, Warsaw University of Life Sciences - SGGW (WULS-SGGW), Warsaw, Poland

\*Address all correspondence to<EMAIL_ADDRESS>
© 2020 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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**89**

*The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria…*

[25] Mozaffarian D. Changes in diet and lifestyle and long-term weight gain in women and men. The New England Journal of Medicine.

[26] Chen M, Sun Q, Giovannucci E, Mozaffarian D, Manson JE, Willett WC, et al. Dairy consumption and risk of type 2 diabetes: 3 cohorts of US adults and an updated metaanalysis. BMC

2011;**364**:2392-2404

Medicine. 2014;**12**(1):215

Nutrition. 2015;**113**:131-135

Polish, Abstract in English)

English)

[27] Tapsell LC. Fermented dairy food and CVD risk. British Journal of

[28] Dziugan P., Dziedziczak K., Ambroziak W.: Błonnik w pieczywie. Cukiernictwo i Piekarstwo. 2006; 10 (05), 60-62 (in Polish, Abstract in

[29] Kowalska E. The influence of lactic acid bacteria and bifidobacteria on selected characteristics of fermented buckwheat beverages. 2019. [thesis] (in

[30] Hassan AA, Aly MM, El-Hadidie ST. Production of cereal-based probiotic beverages. World Applied Sciences Journal. 2012;**19**(10):1367-1380

[31] Champagne CP, Green-Johnson J, Raymond Y, Barrette J, Buckley N. Selection of probiotic bacteria for the fermentation of a soy beverage in combination with Streptococcus thermophilus. Food Research International. 2009;**42**:612-621

[32] Rathore S, Salmerón I, Pandiella SS. Production of potentially probiotic beverages using single and mixed cereal substrates fermented with lactic acid bacteria cultures. Food Microbiology.

[33] Ziarno M, Zaręba D, Maciejak M, Veber AL. The impact of dairy starter cultures on selected qualitative

2012;**30**(1):239-244

*DOI: http://dx.doi.org/10.5772/intechopen.94913*

[17] Egert M, de Graaf A, Smidt H, de Vos W, Venema K. Beyond diversity: functional microbiomics of the human colon. Trends in Microbiology.

[18] Blandon J., Cranfield J., Henson S.: Functional Food and Natural Health Product Issues: The Canadian and International Context. International Food Economy Research Group Department of Food, Agricultural and Resource Economics, University of

2006;**14**:86-91

Guelph, Kanada. 2007

[19] Arihara K., Nakashima Y.,

[20] Sanders ME. Overview of

1998;**8**:341-347

2010;**121**:899-906

WileyBlackwell. 2008

2017;**44**:94-102

functional foods: emphasis on probiotic bacteria. International Dairy Journal.

[21] Granato D, Castro IA, Masson ML, Ribeiro JCB. Sensory evaluation and physicochemical optimization of soy-based desserts using response surface methodology. Food Chemistry.

[22] ADA: Position of the American Dietetic Association: functional foods. Journal of the American Dietetic Association. 2009; 109, 735-46

[23] Hutkins RW. Microbiology and Technology of Fermented Foods.

[24] Marco ML, Heeney D, Binda S, Cifelli CJ, Cotter PD, Foligné B, et al. Health benefits of fermented foods: microbiota and beyond. Current Opinion in Biotechnology.

Ishikawa S., Itoh M.: Antihypertensive activities generated from porcine skeletal muscle proteins by lactic acid bacteria. In: Abstracts of 50th International Congress of Meat Science and Technology; Aug 8-13 2004, Helsinki, Finlandis, Elsevier

*The Possibility of Obtaining Buckwheat Beverages Fermented with Lactic Acid Bacteria… DOI: http://dx.doi.org/10.5772/intechopen.94913*

[17] Egert M, de Graaf A, Smidt H, de Vos W, Venema K. Beyond diversity: functional microbiomics of the human colon. Trends in Microbiology. 2006;**14**:86-91

[18] Blandon J., Cranfield J., Henson S.: Functional Food and Natural Health Product Issues: The Canadian and International Context. International Food Economy Research Group Department of Food, Agricultural and Resource Economics, University of Guelph, Kanada. 2007

[19] Arihara K., Nakashima Y., Ishikawa S., Itoh M.: Antihypertensive activities generated from porcine skeletal muscle proteins by lactic acid bacteria. In: Abstracts of 50th International Congress of Meat Science and Technology; Aug 8-13 2004, Helsinki, Finlandis, Elsevier

[20] Sanders ME. Overview of functional foods: emphasis on probiotic bacteria. International Dairy Journal. 1998;**8**:341-347

[21] Granato D, Castro IA, Masson ML, Ribeiro JCB. Sensory evaluation and physicochemical optimization of soy-based desserts using response surface methodology. Food Chemistry. 2010;**121**:899-906

[22] ADA: Position of the American Dietetic Association: functional foods. Journal of the American Dietetic Association. 2009; 109, 735-46

[23] Hutkins RW. Microbiology and Technology of Fermented Foods. WileyBlackwell. 2008

[24] Marco ML, Heeney D, Binda S, Cifelli CJ, Cotter PD, Foligné B, et al. Health benefits of fermented foods: microbiota and beyond. Current Opinion in Biotechnology. 2017;**44**:94-102

[25] Mozaffarian D. Changes in diet and lifestyle and long-term weight gain in women and men. The New England Journal of Medicine. 2011;**364**:2392-2404

[26] Chen M, Sun Q, Giovannucci E, Mozaffarian D, Manson JE, Willett WC, et al. Dairy consumption and risk of type 2 diabetes: 3 cohorts of US adults and an updated metaanalysis. BMC Medicine. 2014;**12**(1):215

[27] Tapsell LC. Fermented dairy food and CVD risk. British Journal of Nutrition. 2015;**113**:131-135

[28] Dziugan P., Dziedziczak K., Ambroziak W.: Błonnik w pieczywie. Cukiernictwo i Piekarstwo. 2006; 10 (05), 60-62 (in Polish, Abstract in English)

[29] Kowalska E. The influence of lactic acid bacteria and bifidobacteria on selected characteristics of fermented buckwheat beverages. 2019. [thesis] (in Polish, Abstract in English)

[30] Hassan AA, Aly MM, El-Hadidie ST. Production of cereal-based probiotic beverages. World Applied Sciences Journal. 2012;**19**(10):1367-1380

[31] Champagne CP, Green-Johnson J, Raymond Y, Barrette J, Buckley N. Selection of probiotic bacteria for the fermentation of a soy beverage in combination with Streptococcus thermophilus. Food Research International. 2009;**42**:612-621

[32] Rathore S, Salmerón I, Pandiella SS. Production of potentially probiotic beverages using single and mixed cereal substrates fermented with lactic acid bacteria cultures. Food Microbiology. 2012;**30**(1):239-244

[33] Ziarno M, Zaręba D, Maciejak M, Veber AL. The impact of dairy starter cultures on selected qualitative

**88**

*Milk Substitutes - Selected Aspects*

[1] Nowak A., Slizewska K., Libudzisz Z.: Probiotyki-historia i mechanizmy dzialania. Żywność Nauka Technologia Jakość. 2010; 17(4), 5-19 (in

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[2] Turnbull JL, Adams HN, Gorard DA. The diagnosis and management of food allergy and food intolerances. Alimentary Pharmacology and Therapeutics. 2015;**41**(1):3-25

inhibitory activities. Current Advances in Buckwheat Research. 1995;**1**:927-934

[9] Christa K, Soral-Smietana M. Buckwheat grains and buckwheat products nutritional and prophylactic value of their components- a review. Czech Journal of Food Sciences.

[10] Ikeda S, Tomura K, Yamashita Y, Kreft I. Minerals in buckwheat flour subjected to enzymatic digestion. Fagopyrum. 2001;**18**:45-48

*Fagopyrum esculentum* Moench. Kanada: International Plant Genetic Resources

[11] Campbell CG. Buckwheat:

[12] Duffus CM, Binnie J. Sucrose relationships during endosperm and embryo development in wheat. Plant Physiology and Biochemistry.

[13] Kalinova J, Triska J, Vrchotova N. Distribution of vitamin E, squalene, epicatechin and rutin in common buckwheat plants (Fagopyrum esculentum Moench). Journal of Agricultural and Food Chemistry.

[14] Zhang ZL, Zhou ML, Tang Y, Li FL, Tang YX, Shao JR, et al. Bioactive compounds in functional buckwheat food. Food Research International.

[15] Schaafsama G.: State-of-theart concerning probiotic strains in milk products. International Dairy Federation, Nutrition Newsletter. 1996;

[16] Dominguez-Bello MG,

2008;**10**:1072-1076

Blaser MJ. Do you have a probiotic in your future? Microbes an Infection.

2008;**26**:153-162

Institute; 1997

1990;**28**(2):161-165

2006;**54**:5330-5335

2012;**49**(1):389-395

5, 23-24 83

[3] Zopf Y, Baenkler HW, Silbermann A, Hahn EG, Raithel M. The differential

diagnosis of food intolerance. Deutsches Ärzteblatt International.

[4] Lomer MCE. The aetiology, diagnosis, mechanisms and clinical evidence for food intolerance. Alimentary Pharmacology and Therapeutics. 2015;**41**(3):262-275

[5] Hischenhuber C, Crevel R,

[6] Zarzecka K., Marek G.,

Polish, Abstract in English)

[7] Grochowicz J., Dominik P., Fabisiak A. The possibilities of using natural foods as a result of the global

trend in food demand health

prevention. Zeszyty Naukowe Uczelni Vistula. 2017; 54.3. Turystyka III, 223- 240 (in Polish, Abstract in English)

[8] Kawakami A, Inbe T, Kayahara H, Horii A. Preparation of enzymatic hydrolysates of buckwheat globulin and their angiotensin I converting enzyme

Jarry B, Maki M, Moneret-Vautrin DA, Romano A, et al. Review article: Safe amounts of gluten for patients with wheat allergy or coeliac disease. Alimentary Pharmacology and Therapeutics. 2006;**23**:559-575

Iwona M.: Nutritional and pro-health value of buckwheat. Problemy Higieny i Epidemiologii. 2015; 96(2), 410-413 (in

2009;**106**:359-369

properties of functional fermented beverage prepared from germinated White Kidney Beans. Journal of Food and Nutrition Research. 2019;**2**:167-176 ISSN: 1338-4260

[34] Zareba D, Ziarno M, Ścibisz I, Gawron J. The importance of volatile compound profile in the assessment of fermentation conducted by Lactobacillus casei DN-114001. International Dairy Journal. 2014;**35**:11-14

[35] Mituniewicz-Małek A, Ziarno M, Dmytrów I, Balejko J. Effect of the addition of Bifidobacterium monocultures on the physical, chemical and sensory characteristics of fermented goat milk. Journal of Dairy Science. 2017;**100**:6972-6979

[36] Ramos CL, de Almeida EG, Freire AL, Schwan RF. Diversity of bacteria and yeast in the naturally fermented cotton seed and rice beverage produced by Brazilian Amerindians. Food Microbiology. 2011;**28**(7):1380-1386

[37] Němečková I, Dragounova H, Pechačová M, Rysova J, Roubal P. Fermentation of vegetable substrates by lactic acid bacteria as a basis of functional foods. Czech Journal of Food Sciences. 2012;**29**:42-48

[38] FAO/WHO: Codex Alimentarius Commision. Annex Proposed Draft Standard for Fermented Milks. 1997

[39] Santos AD, Auler CC, Da Silva Libeck B, Schwan RF. Co-culture fermentation of peanut-soy milk for the development of a novel functional beverage. International Journal of Food Microbiology. 2014;**186**:32-41

[40] Phiarais NPB, Mauch A, Schehl DB, Zarnkow M, Gastl M, Herrmann M, et al. Processing of a top fermented beer brewed from 100% buckwheat malt with sensory and analytical

characterisation. Journal of the Institute of Brewing. 2010;**116**(3):265-274

[41] Campbell C. G.: Buckwheat: Fagopyrum esculentum Moench. International Plant Genetic Resources Institute. 1997, Canada

[42] Zondag MD. Effect of microwave heat-moisture and annealing treatments on buckwheat starch characteristics. Research Paper: University of Wisconsin, Stout; 2003

[43] Wajcht M. Research on the use of *Propionibacterium* sp. in the production of fermented milk and vegetable drinks, Warsaw University of Life Sciences; 2019. [thesis] (in Polish, Abstract in English)

**91**

**Chapter 6**

**Abstract**

lactic acid bacteria, bifidobacteria

**1. Introduction**

Functional Fermented Beverage

*Anna Veber, Dorota Zaręba and Małgorzata Ziarno*

Prepared from Germinated White

Kidney Beans (*Phaseolus vulgaris* L.)

The current demand for plant-based food indicates that the food market is providing alternatives for products that are currently commercially available. This chapter discusses the possible use of germinated bean seeds as a raw material in the production of substitutes for dairy products, including fermented ones. Beans are a valuable source of easily digestible protein, carbohydrates, minerals, and various vitamins (e.g., B vitamin group). They also contain significant amounts of fiber which affects the proper functioning of the digestive system and antioxidant compounds.  However, it is mainly (about 70%) constituted by unsaturated fatty acids, including the polyunsaturated ones such as linoleic acid or linolenic acid, which are desirable in the human diet for the prevention of cardiovascular diseases or cancer. Biological processes such as germination or fermentation may improve the nutritional value of bean seeds (by increasing the content, digestibility, and bioavailability of some nutrients and by eliminating undesirable components) and deliver live cells of prohealth bacteria (lactic acid bacteria, propionic acid bacteria, or bifidobacteria).

**Keywords:** bean-based beverages, white kidney beans, germination, fermentation,

Common bean (*Phaseolus vulgaris* L.) is an annual angiosperm belonging to the Fabaceae family. It originates from Central and South America, where it was cultivated as early as 7000 years ago. Due to their ability to self-pollinate, beans can produce seeds after pollination with pollen from their own flower [1]. There are about 200 species identified so far, of which the first known were green pods with dark seeds. Based on the evolutionary rate, organisms of the genus *Phaseolus* are estimated to be about six million years old, which suggests that it is a relatively young group of plants [2, 3]. Currently, most varieties of beans are cultivated for

Bean varieties are classified into two main groups: dwarf and tic. In the case of dwarf varieties, a poorly developed stem reaches 60 cm, whereas in climbing varieties, the stem can grow up to 200–300 cm. Features such as shape, color, and size of the pods are related to the variety. There is a relationship between the shape of the seeds and the shape of the pod. Elongated and cylindrical seeds are found in

food, on all continents and in various climatic zones [1].

## **Chapter 6**

*Milk Substitutes - Selected Aspects*

ISSN: 1338-4260

2014;**35**:11-14

[35] Mituniewicz-Małek A,

Science. 2017;**100**:6972-6979

2011;**28**(7):1380-1386

Sciences. 2012;**29**:42-48

[36] Ramos CL, de Almeida EG, Freire AL, Schwan RF. Diversity of bacteria and yeast in the naturally fermented cotton seed and rice beverage produced by Brazilian Amerindians. Food Microbiology.

[37] Němečková I, Dragounova H, Pechačová M, Rysova J, Roubal P. Fermentation of vegetable substrates by lactic acid bacteria as a basis of functional foods. Czech Journal of Food

[38] FAO/WHO: Codex Alimentarius Commision. Annex Proposed Draft Standard for Fermented Milks. 1997

[39] Santos AD, Auler CC, Da Silva Libeck B, Schwan RF. Co-culture fermentation of peanut-soy milk for the development of a novel functional beverage. International Journal of Food

Microbiology. 2014;**186**:32-41

[40] Phiarais NPB, Mauch A, Schehl DB, Zarnkow M, Gastl M, Herrmann M, et al. Processing of a top fermented beer brewed from 100% buckwheat malt with sensory and analytical

Ziarno M, Dmytrów I, Balejko J. Effect of the addition of Bifidobacterium monocultures on the physical,

chemical and sensory characteristics of fermented goat milk. Journal of Dairy

properties of functional fermented beverage prepared from germinated White Kidney Beans. Journal of Food and Nutrition Research. 2019;**2**:167-176 characterisation. Journal of the Institute

of Brewing. 2010;**116**(3):265-274

[41] Campbell C. G.: Buckwheat: Fagopyrum esculentum Moench. International Plant Genetic Resources

[42] Zondag MD. Effect of microwave heat-moisture and annealing treatments on buckwheat starch characteristics. Research Paper: University of Wisconsin, Stout; 2003

[43] Wajcht M. Research on the use of *Propionibacterium* sp. in the production of fermented milk and vegetable drinks, Warsaw University of Life Sciences; 2019. [thesis] (in Polish, Abstract in

Institute. 1997, Canada

English)

[34] Zareba D, Ziarno M, Ścibisz I, Gawron J. The importance of volatile compound profile in the assessment of fermentation conducted by Lactobacillus casei DN-114001. International Dairy Journal.

**90**

## Functional Fermented Beverage Prepared from Germinated White Kidney Beans (*Phaseolus vulgaris* L.)

*Anna Veber, Dorota Zaręba and Małgorzata Ziarno*

## **Abstract**

The current demand for plant-based food indicates that the food market is providing alternatives for products that are currently commercially available. This chapter discusses the possible use of germinated bean seeds as a raw material in the production of substitutes for dairy products, including fermented ones. Beans are a valuable source of easily digestible protein, carbohydrates, minerals, and various vitamins (e.g., B vitamin group). They also contain significant amounts of fiber which affects the proper functioning of the digestive system and antioxidant compounds.  However, it is mainly (about 70%) constituted by unsaturated fatty acids, including the polyunsaturated ones such as linoleic acid or linolenic acid, which are desirable in the human diet for the prevention of cardiovascular diseases or cancer. Biological processes such as germination or fermentation may improve the nutritional value of bean seeds (by increasing the content, digestibility, and bioavailability of some nutrients and by eliminating undesirable components) and deliver live cells of prohealth bacteria (lactic acid bacteria, propionic acid bacteria, or bifidobacteria).

**Keywords:** bean-based beverages, white kidney beans, germination, fermentation, lactic acid bacteria, bifidobacteria

## **1. Introduction**

Common bean (*Phaseolus vulgaris* L.) is an annual angiosperm belonging to the Fabaceae family. It originates from Central and South America, where it was cultivated as early as 7000 years ago. Due to their ability to self-pollinate, beans can produce seeds after pollination with pollen from their own flower [1]. There are about 200 species identified so far, of which the first known were green pods with dark seeds. Based on the evolutionary rate, organisms of the genus *Phaseolus* are estimated to be about six million years old, which suggests that it is a relatively young group of plants [2, 3]. Currently, most varieties of beans are cultivated for food, on all continents and in various climatic zones [1].

Bean varieties are classified into two main groups: dwarf and tic. In the case of dwarf varieties, a poorly developed stem reaches 60 cm, whereas in climbing varieties, the stem can grow up to 200–300 cm. Features such as shape, color, and size of the pods are related to the variety. There is a relationship between the shape of the seeds and the shape of the pod. Elongated and cylindrical seeds are found in round and long pods, while flattened seeds are found in flat pods. The bean fruit is an elongated pod, which varies in color, shape, and fiber content, depending on the variety.

Beans are one of the most important plants that are directly consumed in the world. Due to their nutritional and health benefits, they are used in many dishes and are also consumed by people following vegan and vegetarian diets as a valuable source of vegetable protein. In some regions, such as South and Central American and African countries, beans are a staple in the daily diet and usually consumed after soaking and cooking.

Consuming the seeds of legumes, which include beans, can result in many physiological and health benefits, including the prevention of cardiovascular disease, diabetes, and cancer. Beans are high-fiber, high-protein vegetables that contain a less amount of fats. They are valuable sources of not only easily digestible protein but also minerals and various vitamins (e.g., B vitamins). Furthermore, they contain a wide range of phytochemical and antioxidant compounds, including flavonoids, such as anthocyanins, flavonols, phenolic acids, and isoflavones, which are compounds regulating the expression of genes responsible for the processes of β-fat oxidation, lithogenesis, and hepatic gluconeogenesis. Beans are also rich in oligosaccharides, lectins, saponins, phytates, and polyphenolic compounds, the main classes of which are tannins, phenolic acids, and flavonoids. Phenolic extracts from various types of beans exhibit antioxidant properties [4]. The polyphenolic components present in beans are mainly concentrated in the seed coat and give the seeds their color. Legume seeds also have a lower glycemic index compared to other starchy foods such as cereals and potatoes. When added in the daily diet, legumes can exert many beneficial physiological effects and prevent common metabolic diseases such as diabetes, coronary artery disease, and cancer [5–7]. They are effective in lowering blood pressure, normalizing body weight and lipid metabolism, reducing insulin resistance, and thus regulating the components of the metabolic syndrome and are therefore recommended for its prevention and treatment [8].

Use of the germination of bean seeds and then the fermentation process of beverages obtained from the germinated bean seeds allow to increase the health and nutritional values of these beverages. This chapter presents various studies about technology, quality, and nutritional value of fermented bean-based beverages prepared from germinated white kidney beans (*Phaseolus vulgaris* L.).

## **2. Preparation of beverages from beans**

Plant milk substitutes are obtained mainly by water extraction of selected plant material. The production process is of different types, depending on the raw material used (legumes, cereals, vegetables, nuts, seeds), but all the variants have a common outline. Generally, the preparation process involves the following stages: selection of the raw material, soaking and wet or dry grinding of the raw material, water extraction of the raw material, heating, separation of the solid fraction, cooling, standardization, homogenization, thermal fixation, aseptic packaging, and storage [9–12].

In some cases, additional blanching or roasting of the raw material is carried out prior to soaking and grinding. Blanching is usually done in boiling water for 1–5 minutes, for example, to inactivate trypsin and lipoxygenase inhibitors in the case of soybean beverages. Moreover, it prevents the formation of foam during the technological process. Roasting is carried out at a temperature above 100°C, in hot air, and its duration is determined by the type of the raw material and the temperature of the process. This process is used to improve the taste and aroma of the final

**93**

process [9, 12].

**and propionic acid bacteria**

*Functional Fermented Beverage Prepared from Germinated White Kidney Beans…*

product; however, it may reduce the protein solubility and extraction efficiency [9]. In the case of bean-based beverages, before soaking and grinding the seeds, it is

Soaking and grinding (or only grinding) of the raw material are employed for further processing steps and to facilitate the release of nutrients contained in it. Water inactivates some of the inhibitors and reduces the amount of phytic acid, which increases the absorption of nutrients and their bioavailability [9–14]. In the case of beans, wet grinding is performed after thermal treatment of the seeds to induce starch thermohydrolysis. For some plant materials, enzymes are also added at this stage to induce enzymatic hydrolysis of starch or the polysaccharides present. An example of an enzyme used is alpha-amylase, which catalyzes the hydrolysis of the α-1,4-glycosidic linkage of amylose and amylopectin present in starch and results in the formation of shorter-chain compounds, mainly in the form of dextrins. Proteolytic enzymes are also used for increasing the protein digestibility and extraction efficiency, as well as for improving the suspension stability [9, 15]. Beans can also be subjected to such a process. Alternatively, initial dry grinding of the raw material can be employed, followed by its aqueous extraction at an elevated

The next step in the production of plant-based beverages is the separation of the solid from the liquid fraction, by filtration or centrifugation of the obtained suspension. The resulting plant-based beverage may undergo the standardization process in order to obtain a product with a previously assumed composition, enriched with vitamins or minerals. In the case of bean seeds, subsequent additional heating is carried out to a particular temperature depending on the specificity of the

Usually, ultrahigh temperature (UHT) treatment is applied, where the product is heated in flow to 135–150°C for a few seconds to obtain a commercially sterile one. This process degrades and converts the vegetative forms into microorganisms, while maintaining the taste and aroma of the product. The obtained, microbiologically safe product is poured into unit packages, stored, and distributed [9, 10, 12].

Plant-based beverages exhibit low suspension stability due to the presence of solid particles (e.g., protein, starch, fiber, and other solid residues of plant material). To increase the stability of cow milk substitutes, hydrocolloids of plant origin are added before the final thermal run. Alternatively, the resulting suspension is subjected to homogenization and micronization, which leads to an increase in the stability of the system without the addition of hydrocolloids. This process involves simultaneous grinding and mixing of the particles of the dispersed phase, while forcing the liquid heterogeneous system under high pressure (15–25 MPa) through the homogenizing gap. After micronization, the particle size usually ranges from 0.5 μm to 10 μm. Consequently, the obtained product will have greater creaminess and homogeneity compared to the beverage subjected only to the homogenization

**3. Fermentation of bean-based beverages using lactic acid bacteria**

For many years, attempts have been made to obtain fermented plant-based beverages with the same amount of lactic acid bacteria as in the fermented dairy beverages [16–18]. Several biotechnology companies offer starter cultures for the fermentation of plant products. The addition of these cultures is aimed at obtaining vegan fermented beverages, which are substitutes for milk yogurts. These products most often contain microorganisms that are used for the fermentation of milk,

*DOI: http://dx.doi.org/10.5772/intechopen.95818*

advisable to carry out germination [10, 12].

temperature [9].

raw material [9, 10, 12].

#### *Functional Fermented Beverage Prepared from Germinated White Kidney Beans… DOI: http://dx.doi.org/10.5772/intechopen.95818*

product; however, it may reduce the protein solubility and extraction efficiency [9]. In the case of bean-based beverages, before soaking and grinding the seeds, it is advisable to carry out germination [10, 12].

Soaking and grinding (or only grinding) of the raw material are employed for further processing steps and to facilitate the release of nutrients contained in it. Water inactivates some of the inhibitors and reduces the amount of phytic acid, which increases the absorption of nutrients and their bioavailability [9–14]. In the case of beans, wet grinding is performed after thermal treatment of the seeds to induce starch thermohydrolysis. For some plant materials, enzymes are also added at this stage to induce enzymatic hydrolysis of starch or the polysaccharides present. An example of an enzyme used is alpha-amylase, which catalyzes the hydrolysis of the α-1,4-glycosidic linkage of amylose and amylopectin present in starch and results in the formation of shorter-chain compounds, mainly in the form of dextrins. Proteolytic enzymes are also used for increasing the protein digestibility and extraction efficiency, as well as for improving the suspension stability [9, 15]. Beans can also be subjected to such a process. Alternatively, initial dry grinding of the raw material can be employed, followed by its aqueous extraction at an elevated temperature [9].

The next step in the production of plant-based beverages is the separation of the solid from the liquid fraction, by filtration or centrifugation of the obtained suspension. The resulting plant-based beverage may undergo the standardization process in order to obtain a product with a previously assumed composition, enriched with vitamins or minerals. In the case of bean seeds, subsequent additional heating is carried out to a particular temperature depending on the specificity of the raw material [9, 10, 12].

Usually, ultrahigh temperature (UHT) treatment is applied, where the product is heated in flow to 135–150°C for a few seconds to obtain a commercially sterile one. This process degrades and converts the vegetative forms into microorganisms, while maintaining the taste and aroma of the product. The obtained, microbiologically safe product is poured into unit packages, stored, and distributed [9, 10, 12].

Plant-based beverages exhibit low suspension stability due to the presence of solid particles (e.g., protein, starch, fiber, and other solid residues of plant material). To increase the stability of cow milk substitutes, hydrocolloids of plant origin are added before the final thermal run. Alternatively, the resulting suspension is subjected to homogenization and micronization, which leads to an increase in the stability of the system without the addition of hydrocolloids. This process involves simultaneous grinding and mixing of the particles of the dispersed phase, while forcing the liquid heterogeneous system under high pressure (15–25 MPa) through the homogenizing gap. After micronization, the particle size usually ranges from 0.5 μm to 10 μm. Consequently, the obtained product will have greater creaminess and homogeneity compared to the beverage subjected only to the homogenization process [9, 12].

## **3. Fermentation of bean-based beverages using lactic acid bacteria and propionic acid bacteria**

For many years, attempts have been made to obtain fermented plant-based beverages with the same amount of lactic acid bacteria as in the fermented dairy beverages [16–18]. Several biotechnology companies offer starter cultures for the fermentation of plant products. The addition of these cultures is aimed at obtaining vegan fermented beverages, which are substitutes for milk yogurts. These products most often contain microorganisms that are used for the fermentation of milk,

*Milk Substitutes - Selected Aspects*

after soaking and cooking.

variety.

round and long pods, while flattened seeds are found in flat pods. The bean fruit is an elongated pod, which varies in color, shape, and fiber content, depending on the

Beans are one of the most important plants that are directly consumed in the world. Due to their nutritional and health benefits, they are used in many dishes and are also consumed by people following vegan and vegetarian diets as a valuable source of vegetable protein. In some regions, such as South and Central American and African countries, beans are a staple in the daily diet and usually consumed

Consuming the seeds of legumes, which include beans, can result in many physiological and health benefits, including the prevention of cardiovascular disease, diabetes, and cancer. Beans are high-fiber, high-protein vegetables that contain a less amount of fats. They are valuable sources of not only easily digestible protein but also minerals and various vitamins (e.g., B vitamins). Furthermore, they contain a wide range of phytochemical and antioxidant compounds, including flavonoids, such as anthocyanins, flavonols, phenolic acids, and isoflavones, which are compounds regulating the expression of genes responsible for the processes of β-fat oxidation, lithogenesis, and hepatic gluconeogenesis. Beans are also rich in oligosaccharides, lectins, saponins, phytates, and polyphenolic compounds, the main classes of which are tannins, phenolic acids, and flavonoids. Phenolic extracts from various types of beans exhibit antioxidant properties [4]. The polyphenolic components present in beans are mainly concentrated in the seed coat and give the seeds their color. Legume seeds also have a lower glycemic index compared to other starchy foods such as cereals and potatoes. When added in the daily diet, legumes can exert many beneficial physiological effects and prevent common metabolic diseases such as diabetes, coronary artery disease, and cancer [5–7]. They are effective in lowering blood pressure, normalizing body weight and lipid metabolism, reducing insulin resistance, and thus regulating the components of the metabolic syndrome and are therefore recommended for its prevention and treatment [8]. Use of the germination of bean seeds and then the fermentation process of beverages obtained from the germinated bean seeds allow to increase the health and nutritional values of these beverages. This chapter presents various studies about technology, quality, and nutritional value of fermented bean-based beverages

prepared from germinated white kidney beans (*Phaseolus vulgaris* L.).

Plant milk substitutes are obtained mainly by water extraction of selected plant material. The production process is of different types, depending on the raw material used (legumes, cereals, vegetables, nuts, seeds), but all the variants have a common outline. Generally, the preparation process involves the following stages: selection of the raw material, soaking and wet or dry grinding of the raw material, water extraction of the raw material, heating, separation of the solid fraction, cooling, standardization, homogenization, thermal fixation, aseptic packaging, and

In some cases, additional blanching or roasting of the raw material is carried out prior to soaking and grinding. Blanching is usually done in boiling water for 1–5 minutes, for example, to inactivate trypsin and lipoxygenase inhibitors in the case of soybean beverages. Moreover, it prevents the formation of foam during the technological process. Roasting is carried out at a temperature above 100°C, in hot air, and its duration is determined by the type of the raw material and the temperature of the process. This process is used to improve the taste and aroma of the final

**2. Preparation of beverages from beans**

**92**

storage [9–12].

including lactic acid bacteria and bifidobacteria such as *Streptococcus thermophilus*, *Lactobacillus delbrueckii* subsp. *bulgaricus*, *Lactobacillus acidophilus*, *Lactobacillus paracasei*, *Bifidobacterium animalis*, and *Bifidobacterium lactis* [19]. Furthermore, Wajcht's research [14] proves the possibility of using *Propionibacterium freudenreichii* subsp. *shermanii* for the production of fermented bean-based beverages (obtained from germinated adzuki bean and mung bean seeds). After 24-hour fermentation of bean beverages at 37°C, the pH was in the range of 4.3–4.7, and the population of propionibacteria was not lower than 7 log10 CFU/mL [14]. Which proves that *Propionibacterium freudenreichii* subsp. *shermanii* show the ability to grow and ferment sugars contained in bean-based beverages to an extent no worse than the lactic acid bacteria do.

The population size of these microorganisms throughout the shelf life of such beverages is an important factor. It is one of the indicators of the quality and health-promoting properties of these products. The minimum number of cells in such products should be at least 7 log10 CFU/mL or g for starter bacteria and at least 6 log10 CFU/mL or g for additional microorganisms (e.g., probiotics) [20]. It has already been established that the selection of bacteria used for fermentation and the composition and acidity of the product have a significant impact on the viability of the starter and prebiotic bacteria, as well as on the maintenance of the required bacterial population [18, 21]. Ziarno et al. [10] used two commercially available industrial yogurt starter cultures, namely Yo-Mix 205 LYO (DuPont Danisco; consisting of *S. thermophilus*, *L. delbrueckii* subsp. *bulgaricus, L. acidophilus*, and *B. lactis* with sacharose and maltodextrins as carriers) and ABY-3 Probio-Tec (Chr. Hansen; consisting of *S. thermophilus*, *L. delbrueckii* subsp. *bulgaricus*, and documented probiotic strains *L. acidophilus* La-5 and *B. animalis* subsp. *lactis* BB-12), for the fermentation of bean-based beverages obtained from germinated bean seeds of the "Piękny Jaś Karłowy" variety. In the fermented beverages thus obtained, the level of bacteria of the starter culture and additional microorganisms was, respectively, at least 7 log10 CFU/mL and at least 6 log10 CFU/mL. During 28 days of storage at 6°C, there was a significant reduction in the population of *S. thermophilus*, *L. acidophilus*, and *Bifidobacterium* in the beverages; however, the levels of microorganisms were not below 7 log10 CFU/mL and 6 log10 CFU/mL, respectively. Maciejak [12] noted that the population of starter microorganisms was at the level of 8.1 log10 CFU/mL in the bean beverage fermented with the industrial starter culture ABY-3. Zaręba and Ziarno [18] showed that the fermentation of plant-based beverage matrices (soy, rice, and coconut beverages) is more conducive to the development and survival of streptococci compared to lactobacilli. However, the survival rate of both lactobacilli and lactobacilli depends on the type of plant-based beverages and the starter culture used, as well as the degree of fermentation of the beverage (final pH). Therefore, the starter culture should be carefully selected for the specific type of plant beverage, and the storage temperature of the final product should also be adjusted.

### **4. Nutritional and health value of bean seeds and bean-based beverages**

Depending on the variety, white bean seeds contain approximately 21–23 g of total proteins, approximately 0.8–1.6 g of fat, approximately 60–63 g of total carbohydrates, including approximately 40 g of starch and approximately 15–24 g of dietary fiber per 100 g [22–26].

The nutritional and health value of bean beverages depends on the recipe composition and the amount of bean seeds in the product [10, 12, 14, 19, 27]. Ziarno et al. [10] obtained a bean-based beverage by blending 100 g of presoaked white

**95**

*Functional Fermented Beverage Prepared from Germinated White Kidney Beans…*

plant-based beverages have a comparable protein content.

bean seeds of the "Piękny Jaś Karłowy" variety with 900 g of drinking water. Then, the homogenate was boiled to gelatinize the starch contained in the beans. The obtained mixture was filtered through a sieve and sterilized at 121°C for 20 minutes. Among other components, the protein content of the beverage obtained was estimated at 2.3 g/100 g. The study by Jeske et al. [28] reported that commercial

The main types of proteins found in legume seeds, including bean seeds, are globulins and albumin, which account for 50–70% and 20% of all proteins, respectively [25]. Globulins, which are the dominant fraction of proteins, are referred to as storage proteins. These are stored in membrane-bound organelles, vacuole stores, or protein bodies. In parenchymal cells, globulins survive drying out during seed maturation and undergo proteolysis during germination, providing free amino

The storage proteins of bean seeds, like other legumes, have a relatively low content of methionine (approximately 0.1–0.37 g/100 g), cysteine and cystine (0.23–0.25 g/100 g), and also tryptophan (0.25–0.27 g/100 g); however, they are rich in lysine (1.4–1.6 g/100 g), which is the limiting amino acid [26]. Consuming legumes and cereals together in a ratio of 35:65 significantly improves the quality of the supplied protein, making the meal wholesome with a favorable composition of

Most legume seeds, including the white bean ones, contain a maximum of about 5% fat in dry matter (DM), except chickpeas and soybeans, which contain approximately 15% and 47% of fat, respectively [32]. However, in the fermented beanbased beverages prepared by Maciejak [12], the total fat content was 0.16 g/100 g. The fat level in commercial plant beverages is usually regulated (and increased), for example, by the addition of vegetable oils [19]. According to the research of Jeske et al. [28], commercial plant-based beverages have a comparable fat content as the

The main lipid components of bean seeds are triacylglycerols (TAGs) and phospholipids [33]. The lipid fraction is rich in mono- and polyunsaturated fatty acids (PUFA) (approximately 0.07–0.10 g/100 g and 0.36–0.51 g/100 g, respectively), which constitute approximately 60% of all fatty acids [26, 34]. In some legumes, PUFA are present in the form of linoleic acid (C18:2, included in the omega-6 acid fraction) and α-linolenic acid (C18:3, included in the omega-3 acid fraction). These acids cannot be synthesized by the human body, so they must be constantly supplied in the diet [31]. Among the unsaturated fatty acids, linoleic acid dominates (0.19–0.27 g/100 g), accounting for 21–53% of all fatty acids in beans [24, 29]. The content of linolenic acid is in the range of 026.16–0.23 g/100 g or accounts for 4–22% of the total pool of fatty acids. The seeds of all types of legumes are characterized by a very low content of trans fatty acids, constituting less than 1% [26, 35]. The fat present in legumes does not contain cholesterol [34]. While examining the lipid profile of white bean seeds of the "Piękny Jaś Karłowy" variety, Ziarno et al. [36] showed that the dominant fatty acid was linolenic acid (C18:2 n-6c), amounting to 39.23% of the total pool. Other unsaturated acids present in significant amounts were α-linolenic acid (23.23% of all fatty acids) and oleic acid (17.58% of all fatty acids). The most abundant saturated acids were palmitic acid (12.78% of all fatty acids) and stearic acid (3.68% of all fatty acids). Overall, all unsaturated and saturated acids, respectively, amounted to 81.87% and 18.13% of the total pool of fatty acids. The remaining acids present were about 0.5% or less of all fatty acids. For comparison, Ryan et al. [37] estimated the share of α-linolenic, linoleic, and oleic acid in kidney beans, which amounted to 45.69%, 26.04%, and 11.97% of all fatty acids, respectively. Palmitic acid and stearic acid, respectively, amounted to 14.2% and 1.3% of the total pool. The proportion of unsaturated fatty

*DOI: http://dx.doi.org/10.5772/intechopen.95818*

acids [7, 24, 29, 30].

amino acids [7, 24, 29–31].

bean-based beverages prepared by Maciejak [12].

#### *Functional Fermented Beverage Prepared from Germinated White Kidney Beans… DOI: http://dx.doi.org/10.5772/intechopen.95818*

bean seeds of the "Piękny Jaś Karłowy" variety with 900 g of drinking water. Then, the homogenate was boiled to gelatinize the starch contained in the beans. The obtained mixture was filtered through a sieve and sterilized at 121°C for 20 minutes. Among other components, the protein content of the beverage obtained was estimated at 2.3 g/100 g. The study by Jeske et al. [28] reported that commercial plant-based beverages have a comparable protein content.

The main types of proteins found in legume seeds, including bean seeds, are globulins and albumin, which account for 50–70% and 20% of all proteins, respectively [25]. Globulins, which are the dominant fraction of proteins, are referred to as storage proteins. These are stored in membrane-bound organelles, vacuole stores, or protein bodies. In parenchymal cells, globulins survive drying out during seed maturation and undergo proteolysis during germination, providing free amino acids [7, 24, 29, 30].

The storage proteins of bean seeds, like other legumes, have a relatively low content of methionine (approximately 0.1–0.37 g/100 g), cysteine and cystine (0.23–0.25 g/100 g), and also tryptophan (0.25–0.27 g/100 g); however, they are rich in lysine (1.4–1.6 g/100 g), which is the limiting amino acid [26]. Consuming legumes and cereals together in a ratio of 35:65 significantly improves the quality of the supplied protein, making the meal wholesome with a favorable composition of amino acids [7, 24, 29–31].

Most legume seeds, including the white bean ones, contain a maximum of about 5% fat in dry matter (DM), except chickpeas and soybeans, which contain approximately 15% and 47% of fat, respectively [32]. However, in the fermented beanbased beverages prepared by Maciejak [12], the total fat content was 0.16 g/100 g. The fat level in commercial plant beverages is usually regulated (and increased), for example, by the addition of vegetable oils [19]. According to the research of Jeske et al. [28], commercial plant-based beverages have a comparable fat content as the bean-based beverages prepared by Maciejak [12].

The main lipid components of bean seeds are triacylglycerols (TAGs) and phospholipids [33]. The lipid fraction is rich in mono- and polyunsaturated fatty acids (PUFA) (approximately 0.07–0.10 g/100 g and 0.36–0.51 g/100 g, respectively), which constitute approximately 60% of all fatty acids [26, 34]. In some legumes, PUFA are present in the form of linoleic acid (C18:2, included in the omega-6 acid fraction) and α-linolenic acid (C18:3, included in the omega-3 acid fraction). These acids cannot be synthesized by the human body, so they must be constantly supplied in the diet [31]. Among the unsaturated fatty acids, linoleic acid dominates (0.19–0.27 g/100 g), accounting for 21–53% of all fatty acids in beans [24, 29]. The content of linolenic acid is in the range of 026.16–0.23 g/100 g or accounts for 4–22% of the total pool of fatty acids. The seeds of all types of legumes are characterized by a very low content of trans fatty acids, constituting less than 1% [26, 35]. The fat present in legumes does not contain cholesterol [34]. While examining the lipid profile of white bean seeds of the "Piękny Jaś Karłowy" variety, Ziarno et al. [36] showed that the dominant fatty acid was linolenic acid (C18:2 n-6c), amounting to 39.23% of the total pool. Other unsaturated acids present in significant amounts were α-linolenic acid (23.23% of all fatty acids) and oleic acid (17.58% of all fatty acids). The most abundant saturated acids were palmitic acid (12.78% of all fatty acids) and stearic acid (3.68% of all fatty acids). Overall, all unsaturated and saturated acids, respectively, amounted to 81.87% and 18.13% of the total pool of fatty acids. The remaining acids present were about 0.5% or less of all fatty acids. For comparison, Ryan et al. [37] estimated the share of α-linolenic, linoleic, and oleic acid in kidney beans, which amounted to 45.69%, 26.04%, and 11.97% of all fatty acids, respectively. Palmitic acid and stearic acid, respectively, amounted to 14.2% and 1.3% of the total pool. The proportion of unsaturated fatty

*Milk Substitutes - Selected Aspects*

than the lactic acid bacteria do.

including lactic acid bacteria and bifidobacteria such as *Streptococcus thermophilus*, *Lactobacillus delbrueckii* subsp. *bulgaricus*, *Lactobacillus acidophilus*, *Lactobacillus paracasei*, *Bifidobacterium animalis*, and *Bifidobacterium lactis* [19]. Furthermore, Wajcht's research [14] proves the possibility of using *Propionibacterium freudenreichii* subsp. *shermanii* for the production of fermented bean-based beverages (obtained from germinated adzuki bean and mung bean seeds). After 24-hour fermentation of bean beverages at 37°C, the pH was in the range of 4.3–4.7, and the population of propionibacteria was not lower than 7 log10 CFU/mL [14]. Which proves that *Propionibacterium freudenreichii* subsp. *shermanii* show the ability to grow and ferment sugars contained in bean-based beverages to an extent no worse

The population size of these microorganisms throughout the shelf life of such

**4. Nutritional and health value of bean seeds and bean-based beverages**

Depending on the variety, white bean seeds contain approximately 21–23 g of total proteins, approximately 0.8–1.6 g of fat, approximately 60–63 g of total carbohydrates, including approximately 40 g of starch and approximately 15–24 g

The nutritional and health value of bean beverages depends on the recipe composition and the amount of bean seeds in the product [10, 12, 14, 19, 27]. Ziarno et al. [10] obtained a bean-based beverage by blending 100 g of presoaked white

beverages is an important factor. It is one of the indicators of the quality and health-promoting properties of these products. The minimum number of cells in such products should be at least 7 log10 CFU/mL or g for starter bacteria and at least 6 log10 CFU/mL or g for additional microorganisms (e.g., probiotics) [20]. It has already been established that the selection of bacteria used for fermentation and the composition and acidity of the product have a significant impact on the viability of the starter and prebiotic bacteria, as well as on the maintenance of the required bacterial population [18, 21]. Ziarno et al. [10] used two commercially available industrial yogurt starter cultures, namely Yo-Mix 205 LYO (DuPont Danisco; consisting of *S. thermophilus*, *L. delbrueckii* subsp. *bulgaricus, L. acidophilus*, and *B. lactis* with sacharose and maltodextrins as carriers) and ABY-3 Probio-Tec (Chr. Hansen; consisting of *S. thermophilus*, *L. delbrueckii* subsp. *bulgaricus*, and documented probiotic strains *L. acidophilus* La-5 and *B. animalis* subsp. *lactis* BB-12), for the fermentation of bean-based beverages obtained from germinated bean seeds of the "Piękny Jaś Karłowy" variety. In the fermented beverages thus obtained, the level of bacteria of the starter culture and additional microorganisms was, respectively, at least 7 log10 CFU/mL and at least 6 log10 CFU/mL. During 28 days of storage at 6°C, there was a significant reduction in the population of *S. thermophilus*, *L. acidophilus*, and *Bifidobacterium* in the beverages; however, the levels of microorganisms were not below 7 log10 CFU/mL and 6 log10 CFU/mL, respectively. Maciejak [12] noted that the population of starter microorganisms was at the level of 8.1 log10 CFU/mL in the bean beverage fermented with the industrial starter culture ABY-3. Zaręba and Ziarno [18] showed that the fermentation of plant-based beverage matrices (soy, rice, and coconut beverages) is more conducive to the development and survival of streptococci compared to lactobacilli. However, the survival rate of both lactobacilli and lactobacilli depends on the type of plant-based beverages and the starter culture used, as well as the degree of fermentation of the beverage (final pH). Therefore, the starter culture should be carefully selected for the specific type of plant beverage, and the storage temperature of the final product

**94**

should also be adjusted.

of dietary fiber per 100 g [22–26].

acids was 83.8% and that of saturated fatty acids was 16.5%. In another work [38], the fatty acid profile was determined for beans (*P. vulgaris*) and other legumes. For common beans, the content of linoleic acid, α-linolenic acid, and oleic acid (C18:1 n-9) was, respectively, 43.1%, 12.4%, and 13.9% of all fatty acids. The following saturated fatty acids were found in the highest content: palmitic C16:0 (16.8% of all fatty acids) and stearic acids C18:0 (3.5% of all fatty acids). In Adzuki bean seeds, the dominant fatty acids were palmitic, stearic, oleic, linoleic, and α-linolenic acids. Of these, unsaturated fatty acids, mainly linoleic, α-linolenic, and oleic acids, were found in large proportions, which constituted from 70.6% to 73.8% of the total content of fatty acids depending on the variety [34]. Another type of legume seeds studied were mung bean seeds, in which the total amount of lipids was 1.2–1.56% of dry weight depending on the cultivar. As in the previous case, the dominant fatty acid was linoleic acid, which was in the range of 340.5–465.7 mg/100 g of dry weight, while there were significant amounts of palmitic, oleic, α-linolenic, and stearic acids as well [39].

In addition, the positional distribution of fatty acids in the TAG molecules has found interest in increasing research works. Significant differences have been observed in the distribution of fatty acids in TAGs depending on the variety of beans and other legume seeds [33, 37, 38, 40, 41]. In his diploma thesis, Ziarno et al. [36] described the positional distribution of fatty acids in the TAG molecules in the lipids of bean seeds of the "Piękny Jaś Karłowy" variety (**Table 1**). Ryan et al. [37] determined the content of α-linolenic, linoleic, and oleic acids in kidney beans to be, respectively, 45.69%, 26.04%, and 11.97% of all fatty acids. Palmitic acid and stearic acid, respectively, amounted to 14.2% and 1.3% of the total pool of fatty acids. Unsaturated fatty acids amounted to 83.8% of all fatty acids, and saturated acids to 16.5%. In another study, Grela and Gunter [38] determined the fatty acid profile for beans (*P. vulgaris*) and other legumes. In common beans, linoleic acid, α-linolenic acid, and oleic acid amounted to 43.1%, 12.4%, and 13.9% of all fatty acids, respectively. Among the saturated fatty acids, palmitic (16.8% of all fatty acids) and stearic (3.5% of all fatty acids) acids were found in the highest content.

For comparison, in Adzuki bean seeds, most of the unsaturated fatty acids (>96% of fatty acids) were accumulated in the sn-2 position of the TAG molecules. Saturated acids were accumulated in the sn-1 and sn-3 positions, except oleic acid, which was accumulated evenly in all three positions [34]. As for the positional distribution of the TAG molecules in broad bean seeds, similarly to Adzuki beans, a significant amount of unsaturated fatty acids (>95% of fatty acids) were accumulated in the sn-2 position. Only oleic acid was found to be almost evenly occupying the sn-1, sn-2, and sn-3 positions. Saturated acids, such as palmitic and stearic

#### **Table 1.**

*Positional distribution of fatty acids in triacylglycerol (TAG) molecules in the lipids of the white bean seeds of the "Pi*ę*kny Ja*ś *Kar*ł*owy" variety (based on [36]).*

**97**

stomach [24, 43].

was recorded for verbascose.

*Functional Fermented Beverage Prepared from Germinated White Kidney Beans…*

acids, were accumulated in the sn-1 and sn-3 positions. An almost identical distribution was noted in peas, where more than 90% of unsaturated fatty acids were accumulated in the sn-2 position, while saturated acids were mainly accumulated in

The carbohydrates contained in legume seeds, including bean seeds, are mainly composed of starch, fiber, nonstarch polysaccharides, and nondigestible oligosaccharides, which together constitute 30–40% of the dry weight of seeds in the case of species with a high protein content (e.g., lupines and soybeans) or 50–65% of DM in those containing less protein in seeds [25, 42]. However, in the obtained unfermented bean beverages, Cichońska [19] determined a total carbohydrate content of

Bean seeds contain more than 40% of starch and 10–25% of dietary fiber [26]. They are also rich in the resistant fraction of starch, which is not hydrolyzed in the small intestine but is fermented by colonic microorganisms at different rates [7]. The ratio of soluble fiber to insoluble fiber is similar to that found in cereals, which is approximately 1:3 [35]. Consumption of bean fiber is associated with many health and physiological benefits, including improvement in the metabolism of glucose and lipids [24, 43]. When resistant starch in beans is fermented, short-chain fatty acids (such as acetic, butyric, and propionic acid) are produced, the concentration and distribution of which depend on the microorganisms and the carbohydrate content in the gut. Therefore, resistant starch is often considered a prebiotic component as it can regulate the amount and activity of the intestinal flora. It is a precursor of butyrates, which are known for their anti-inflammatory and anticancer properties [44]. In addition, resistant starch binds with bile acids, lowers the absorption of cholesterol and fat, and reduces the absorption of glucose. It also influences the acceleration and regulation of intestinal peristalsis, prevents constipation, and supports the development of beneficial microflora in the large intestine. Furthermore, it reduces hunger and increases satiety after a meal, as it swells in the

Consuming legume seeds, including bean seeds, can cause gas production due to bacterial fermentation, including undigested leftovers. The main oligosaccharide found in legumes is raffinose, which is attributed to the properties of a prebiotic (fermented by probiotic bacteria to short-chain fatty acids); thus, legumes can

Proper preparation of bean beverages can reduce the content of nonenzymatically decomposed ingredients in the human digestive tract [10, 13, 14]. Studies have shown that when lactic acid bacteria are used for the fermentation of legume seeds, the level of stachyose is reduced, which in turn reduces digestive discomfort and flatulence [45–49]. Maciejak [12] obtained fermented and unfermented bean beverages from the germinated white bean seeds of the "Piękny Jaś Karłowy" variety (**Table 2**). Their data showed a significant reduction in the content of all analyzed saccharides. For instance, the content of sucrose was reduced by 82.46%, and raffinose content reached 82.84%. The content of stachyose after the fermentation process was reduced by 68.64%, while the smallest change, amounting to 60.61%,

Of all the seeds of legumes, beans have the highest content of minerals. They can act as an important source of iron (7–11 mg/100 g), zinc (3–4 mg/100 g), copper (0.6–1.0 mg/100 g), phosphorus (300–450 mg/100 g), and potassium (1500–1800 mg/100 g) in the daily diet [23, 24, 26, 50, 51]. Although the content of minerals varies depending on the variety of beans [23, 24], the seeds of white beans are identified as a source of calcium (170–240 mg/100 g) and magnesium (180–190 mg/100 g) [26, 44]. In bean-based beverages, the content of minerals is

improve colon health and reduce the risk of colon cancer [31].

*DOI: http://dx.doi.org/10.5772/intechopen.95818*

the sn-1 and sn-3 positions [33, 40, 41].

1.8–3.7 g/100 g depending on the production recipe used.

#### *Functional Fermented Beverage Prepared from Germinated White Kidney Beans… DOI: http://dx.doi.org/10.5772/intechopen.95818*

acids, were accumulated in the sn-1 and sn-3 positions. An almost identical distribution was noted in peas, where more than 90% of unsaturated fatty acids were accumulated in the sn-2 position, while saturated acids were mainly accumulated in the sn-1 and sn-3 positions [33, 40, 41].

The carbohydrates contained in legume seeds, including bean seeds, are mainly composed of starch, fiber, nonstarch polysaccharides, and nondigestible oligosaccharides, which together constitute 30–40% of the dry weight of seeds in the case of species with a high protein content (e.g., lupines and soybeans) or 50–65% of DM in those containing less protein in seeds [25, 42]. However, in the obtained unfermented bean beverages, Cichońska [19] determined a total carbohydrate content of 1.8–3.7 g/100 g depending on the production recipe used.

Bean seeds contain more than 40% of starch and 10–25% of dietary fiber [26]. They are also rich in the resistant fraction of starch, which is not hydrolyzed in the small intestine but is fermented by colonic microorganisms at different rates [7]. The ratio of soluble fiber to insoluble fiber is similar to that found in cereals, which is approximately 1:3 [35]. Consumption of bean fiber is associated with many health and physiological benefits, including improvement in the metabolism of glucose and lipids [24, 43]. When resistant starch in beans is fermented, short-chain fatty acids (such as acetic, butyric, and propionic acid) are produced, the concentration and distribution of which depend on the microorganisms and the carbohydrate content in the gut. Therefore, resistant starch is often considered a prebiotic component as it can regulate the amount and activity of the intestinal flora. It is a precursor of butyrates, which are known for their anti-inflammatory and anticancer properties [44]. In addition, resistant starch binds with bile acids, lowers the absorption of cholesterol and fat, and reduces the absorption of glucose. It also influences the acceleration and regulation of intestinal peristalsis, prevents constipation, and supports the development of beneficial microflora in the large intestine. Furthermore, it reduces hunger and increases satiety after a meal, as it swells in the stomach [24, 43].

Consuming legume seeds, including bean seeds, can cause gas production due to bacterial fermentation, including undigested leftovers. The main oligosaccharide found in legumes is raffinose, which is attributed to the properties of a prebiotic (fermented by probiotic bacteria to short-chain fatty acids); thus, legumes can improve colon health and reduce the risk of colon cancer [31].

Proper preparation of bean beverages can reduce the content of nonenzymatically decomposed ingredients in the human digestive tract [10, 13, 14]. Studies have shown that when lactic acid bacteria are used for the fermentation of legume seeds, the level of stachyose is reduced, which in turn reduces digestive discomfort and flatulence [45–49]. Maciejak [12] obtained fermented and unfermented bean beverages from the germinated white bean seeds of the "Piękny Jaś Karłowy" variety (**Table 2**). Their data showed a significant reduction in the content of all analyzed saccharides. For instance, the content of sucrose was reduced by 82.46%, and raffinose content reached 82.84%. The content of stachyose after the fermentation process was reduced by 68.64%, while the smallest change, amounting to 60.61%, was recorded for verbascose.

Of all the seeds of legumes, beans have the highest content of minerals. They can act as an important source of iron (7–11 mg/100 g), zinc (3–4 mg/100 g), copper (0.6–1.0 mg/100 g), phosphorus (300–450 mg/100 g), and potassium (1500–1800 mg/100 g) in the daily diet [23, 24, 26, 50, 51]. Although the content of minerals varies depending on the variety of beans [23, 24], the seeds of white beans are identified as a source of calcium (170–240 mg/100 g) and magnesium (180–190 mg/100 g) [26, 44]. In bean-based beverages, the content of minerals is

*Milk Substitutes - Selected Aspects*

stearic acids as well [39].

**Type of fatty acid TAG content of**

*the "Pi*ę*kny Ja*ś *Kar*ł*owy" variety (based on [36]).*

**fatty acid [% of fatty acids]**

Palmitic (C16:0) 12.78 8.68 14.83 22.63 Stearic (C18:0) 3.68 2.16 4.44 19.55 Oleic (C18:1 n-9c) 17.58 16.79 17.97 31.83 Linoleic (C18:2 n-6c) 39.23 46.52 35.59 39.53 α-Linolenic (C18:3 n-3c) 23.23 22.67 23.51 32.53

*Positional distribution of fatty acids in triacylglycerol (TAG) molecules in the lipids of the white bean seeds of*

**Content of fatty acid [% of fatty acids] in position**

**[% of fatty acids] sn-2 sn-1,3**

**Content of fatty acid in the sn-2 position**