Chapter 3

Vol. 4. Cham: Springer; 2018.

Research Trends and Challenges in Smart Grids

Proceedings of the IEEE. 2018;106(1):

[22] Cui J, Liew LS, Sabaliauskaite G, Zhou F. A review on safety failures, security attacks, and available countermeasures for autonomous vehicles. Ad Hoc Networks. 2018. DOI:

10.1016/j.adhoc.2018.12.006

160-170

[15] Rodier C. Travel Effects and Associated Greenhouse Gas Emissions of Automated Vehicles. White paper from The National Center for Sustainable Transportation, Institute for Transportation Studies, UC Davis;

[16] Stephens TS, Gonder J, Chen Y, Lin Z, Liu C, Gohlke D. Estimated bounds and important factors for fuel use and consumer costs of connected and automated vehicles. Technical Report NREL/TP-5400-67216. National Renewable Energy Laboratory, CO,

[17] Stephens TS, Auld J, Chen Y, Gonder J, Kontou E, Lin Z, et al. Assessing energy impacts of connected and automated vehicles at the U.S. National Level—Preliminary bounds and proposed methods. In: Meyer G, Beiker S, editors. Road Vehicle

Automation, Lecture Notes in Mobility.

[18] Wadud Z, MacKenzie D, Leiby P. Help or hindrance? The travel, energy and carbon impact of highly automated vehicles. Transportation Research Part

[19] Petit J, Shladover SE. Potential cyberattacks on automated vehicles. IEEE Transactions on Intelligent Transportation Systems. 2015;16(2):

[20] Parkinson S, Ward P, Wilson K, Miller M. Cyber threats facing autonomous and connected vehicles: Future challenges. IEEE Transactions on Intelligent Transportation Systems.

[21] Karnouskos S, Kerschbaum F. Privacy and integrity considerations in hyperconnected autonomous vehicles.

2017;18(11):2898-2915

Vol. 5. Cham: Springer; 2019.

pp. 113-123

2018

USA; 2016

pp. 105-115

A. 2016;86:1-18

546-556

28


**5. Common oncogenic features exhibited by environmental carcinogens**

The landscape of the genomic disruptions induced by environmental carcinogens is extensive. It has been demonstrated that these compounds can induce alterations such as chromosomal abnormalities, DNA double-strand breaks, gene expression dysregulation, and epigenetic aberrations. While each agent presents a unique mechanism and clinical challenge, a number of parallels can be seen. The molecular effects of exposure to arsenic, asbestos, and radon converge in that each compound can result in DNA damage induced by ROS and inflammation. As these events occur early during tumor development, the identification of the underlying genomic and epigenomic abnormalities caused by these compounds is extremely relevant in identifying early oncogenic events and individual susceptibility differences.

Although the intricacies of the molecular mechanisms of alteration may differ between the various toxic agents, ROS generation is a common outcome of exposure that can lead to extensive DNA damage and further perturbations in various cellular compartments and processes [119]. As mitochondria are one of the primary sources of ROS, they are also key targets of oxidative toxicity [120]. Arsenic exposure is associated with dysfunction of the mitochondria, through the ability of its metabolites to disrupt the mitochondrial membrane potential and reduce mitochondrial ATP levels, as well as ROS-induced mitochondrial damage [121, 122]. Mitochondrial damage induced by arsenic can then lead to numerous alterations in key signaling pathways, such as the decreased expression of apoptotic regulator protein Bcl-2 [122]. Regardless of the molecular mechanism, mitochondrial insult culminates in apoptosis and increased inflammation, in addition to the exacerbation of reactive species generation; events that commonly precede tumorigenesis [121, 123].

Another frequently observed early consequence of exposure to environmental carcinogens is an inflammatory response. Indeed, inflammation caused by infiltrating immune cells underlies numerous hallmarks of cancer biology by providing key molecules for tumor survival and growth, as well as the promotion of genomic aberrations, again through the generation of ROS [124]. Asbestos-induced carcinogenesis is thought to rely heavily on the inflammatory response, where the macrophages of the innate immune system attempt to clear the carcinogenic fibers through phagocytosis [125]. However, these fibers are inherently difficult to digest, leading to the eventual death of the macrophage and subsequent release of proinflammatory cytokines, ROS, and other growth factors [126]. Interestingly, many malignancies have noticeable local immune responses prior to tumor development, highlighting the complex and dichotomous role of host immune cells in both proand anti-tumor functions [127]. Thus, exposure to environmental carcinogens threatens the genetic and epigenetic landscape of oncogene expression in the development of malignancies, and subsequently changes cellular and systemic processes.

The intertwined role of genetic and epigenetic aberrations resulting from exposure to these compounds highlights the complexity of environmentally-induced lung cancer. However, the carcinogenic mechanisms associated with exposure to these agents have been mainly identified using a "one-agent-at-a-time" approach. Further, we have yet to understand how these factors interplay with one another in cases of

**83**

*Oncogenetics of Lung Cancer Induced by Environmental Carcinogens*

combined exposure and how individual genomes modulate the molecular events that arise following exposure. For example, it is difficult to accurately assess the relative risk of lung cancer in an individual who is exposed to occupational asbestos, arsenic-contaminated water, and high levels of domestic indoor air radon. Whether these factors synergize in terms of their molecular effects is not clearly understood and has critical implications to patient monitoring and disease management.

Recently, the idea of the human exposome has sought to provide a method for analyzing individual risk factors by integrating the effects of factors ranging from DNA-level alterations to geographic location. The human exposome is defined as the sum of every exposure to which an individual is subjected to from conception to death [128]. The exposome is dynamic: the nature, amount, and conditions of exposure change over time. It also includes exposure from internal (e.g., metabolism, endogenous hormones, gut microflora, inflammation, oxidative stress, etc.) and external (e.g., radiation, infectious agents, chemical contaminants and environmental pollutants, among others) sources [129]. The lungs are one of the organs at the highest risk of disease development from environmental exposures as the lung exposome can be comprised of an array of molecules and environmental insults. Arsenic, asbestos, and radon, together with air pollution and tobacco smoke, constitute a fraction of the complex mix of environmental carcinogens posing risks for developing thoracic malignancies in humans. However, understanding the oncogenic events following exposure to these agents may allow for the identification of key intervention points to minimize environmentally-induced lung cancer in

**6. Translational outlook for environmentally-induced cancer**

As the molecular mechanisms of environmentally-induced carcinogenesis continue to emerge, a need to characterize the clinical utility of these findings should be underscored. This need is further emphasized by the complex interplay between the numerous features of the lung exposome. Many of the single cancer-associated genes that are affected by exposure to these environmental agents are promising therapeutic intervention points. For instance, targeted inhibitors of EGFR (e.g., erlotinib, afatinib)—a protein transcribed from a gene commonly up-regulated upon exposure to arsenic—are used in lung cancer treatment to interfere with the aberrant growth pathways activated by the upregulation of this signaling receptor [130]. Additionally, the association between radon exposure and *ALK* gene rearrangements in lung cancer patients may be amenable to therapy with inhibitors of the ALK protein (e.g., crizotinib, ceritinib) [131]. However, patients that do not present with alterations in genes that are clinically actionable remain extremely difficult to treat beyond standard regimes. Thus, it is critical to analyze the oncogenetic alterations induced by environmental carcinogens, to not only identify the contribution of each of these widely-distributed agents to tumorigenesis, but also to direct the development of novel treatment and risk-management strategies. Concurrent analysis of altered genes, transcripts, and proteins may help to parse out the risk associated with the varying molecular aberrations that have been observed to be induced by these compounds [132]. This approach, while difficult in terms of scale, necessitates the use of geographic, demographic, and exposome level data, which can be scarce in areas where environmental carcinogen levels are especially concerning. **Table 1** summarizes the currently available sources of information for carcinogens found in the environment that are associated with lung cancer. Overall, future mitigation of the environmental risk factors that lead to lung cancer will rely on the integration of information from the genomic to epidemiological levels.

*DOI: http://dx.doi.org/10.5772/intechopen.81064*

at-risk populations.


*Article* **Scabertopin Derived from** *Elephantopus scaber* **L. Mediates Necroptosis by Inducing Reactive Oxygen Species Production in Bladder Cancer In Vitro**

**Yuanhui Gao 1,†, Zhenyu Nie 1,†, Hui Cao 1, Denggao Huang 1, Mei Chen 1, Yang Xiang 1, Xiaolong Yu 2,\* and Shufang Zhang 1,\***

**Simple Summary:** Scabertopin is one of the major sesquiterpene lactones from *Elephantopus scaber* L. Sesquiterpene lactones are thought to have fairly strong anti-cancer efficacy. However, there has not been any research report on the efficacy and mechanism of Scabertopin in the treatment of bladder cancer. The aim of this study is to evaluate the antitumor activity of scabertopin in bladder cancer and its potential molecular mechanism in vitro. In this study, we found that scabertopin can induce necroptosis in bladder cancer cells by promoting the production of mitochondrial reactive oxygen species, and also inhibit the migration and invasion ability of bladder cancer cells. At the same time, we also demonstrated that the half-inhibition rate of scabertopin on various bladder cancer cell lines is much lower than that on human ureteral epithelial immortalized cells. This shows that scabertopin is a safe and effective anti-bladder cancer drug.

**Abstract:** Bladder cancer remains one of the most common malignant tumors that threatens human health worldwide. It imposes a heavy burden on patients and society due to the high medical costs associated with its easy metastasis and recurrence. Although several treatment options for bladder cancer are available, their clinical efficacy remains unsatisfactory. Therefore, actively exploring new drugs and their mechanisms of action for the clinical treatment of bladder cancer is very important. Scabertopin is one of the major sesquiterpene lactones found in *Elephantopus scaber* L. Sesquiterpene lactones are thought to have fairly strong anti-cancer efficacy. However, the anticancer effect of sesquiterpenoid scabertopin on bladder cancer and its mechanism are still unclear. The aim of this study is to evaluate the antitumor activity of scabertopin in bladder cancer and its potential molecular mechanism in vitro. Our results suggest that scabertopin can induce RIP1/RIP3-dependent necroptosis in bladder cancer cells by promoting the production of mitochondrial reactive oxygen species (ROS), inhibit the expression of MMP-9 by inhibiting the FAK/PI3K/Akt signaling pathway, and ultimately inhibit the migration and invasion ability of bladder cancer cells. At the same time, we also demonstrated that the half-inhibition concentration (IC50) of scabertopin on various bladder cancer cell lines (J82, T24, RT4 and 5637) is much lower than that on human ureteral epithelial immortalized cells (SV-HUC-1). The above observations indicate that scabertopin is a potential therapeutic agent for bladder cancer that acts by inducing necroptosis and inhibiting metastasis.

**Keywords:** scabertopin; bladder cancer; reactive oxygen species; necroptosis; *Elephantopus scaber* L.


Sustainable Buildings from the Perspective Environmental Bioethics

*Nesrin Çobanoğlu*

#### **Abstract**

Today, modern cities are one of the main reasons for current environmental problems. For the solution of these problems, it appears as a necessity to transform the buildings, which are the building blocks of cities, into sustainable structures and also to assign environmental bioethical responsibilities on both individual and society levels. In terms of sustainability, it is important to understand that only a human-centered perspective will not be sufficient; therefore, while constructing and using sustainable buildings, the designing should be based on an environmental bioethics value system. Thus, while constructing sustainable buildings, the environmental bioethics value system should be taken as basis, as well as conservation of energy and human needs. For a sustainable life, sustainable buildings are required within the scope of biopolitics created by adopting environmental bioethical values. Sustainability generally refers to the dynamic situations where systematic needs can be fulfilled in the long term. However, in the ecological context, sustainability can be used to define healthy, self-renewable, no waste-generating, durable, and flexible ecosystems. A sustainable socio-ecological system must include acceptable norms in terms of ethics in addition to these characteristics.

**Keywords:** environmental bioethics, sustainable building, ethics, future generations, sustainability

#### **1. Introduction**

 In general, environmental bioethics is the systematic study of moral relations between people and their natural environment. The theory of environmental ethics has to explain what these rules are, to whom and to what people have responsibilities, and to demonstrate how these responsibilities are justified. These buildings have ecological responsibility and resource productivity throughout their life cycle from design to construction, operation to maintenance, and renewal to destruction. Those buildings which are designed on the principle of minimizing the solid, liquid, and gas wastes as well as the renewable consumption of resources such as energy, water, and building materials can be regarded as sustainable buildings. When constructing sustainable buildings, it is important to design them within the framework of environmental bioethical values.

#### **2. Environmental bioethics**

While trying to find answers to open-ended questions in human-to-human relations, the ethics makes assessments based on "good" and "bad," limiting the responsibility aspect, which is an important component of it, only to human beings. Based on the bioethical concept, the scope of human responsibility covers all living beings such as animals and plants, whereas in terms of environmental ethics, this scope is extended to the relationship with the ecosystem with all its components such as air and water [1].

 In the past, the philosophy of values was used to examine the dilemmas in a narrow space and a narrow period of time, whereas, today, with the reflection of unlimited, fast, and apparent developments in technology, future generations and universal concepts have also appeared as the determinants of those dilemmas. Environmental bioethics should be closely related to social policies, environmental problems, and bioethical concepts.

 There are two main approaches in the environmental ethics. The humancentered "anthropocentric" perspective values the ecosystem as a means of human benefit; on the other hand, the environmental ethics, with an "ecocentric" point of view that adopts the ecosystem as an objective and considers it as the central fact, places the responsibility on human beings while subjectivating the environment due to its own value. They used to defend that the value system was only valid for humans as living beings who think and who are responsible for their behaviors and the subject of values could not be anything other than humans; therefore, living beings other than humans (environment, animals, air, ecosystem as a whole, etc.) could not exist within our world of values due to their values resulting from their own existence and adopted as an objective. Within the scope of environmental ethics, it is defended that, as a result of intellectual developments, the ecosystem is the subject of ethics, not only because of human benefit but also because of its inherent value. Accordingly, the concept of environmental bioethics examines the ethical issues that arise in the network of relations between all living beings and the ecosystem [1–3].

 As in other areas, in environmental terms, the right action is the result of knowledge that follows the interest. "Sliding on ice," which is a bioethical term, should also be considered in terms of environmental ethics. (Sliding on ice is a concept of bioethics that expresses the possibility that the steps we take in good faith toward the right action may lead to catastrophic consequences for the humanity and future generations in cases where we cannot predict all the consequences of the action.) Lack of knowledge or interest results in environmental disasters. The birth of environmental ethics, in fact, is a result of human beings' efforts to survive.

Environmental ethics can be addressed at the universal, professional, and individual levels as the activities of states and countries. In this era, people are unable to fully utilize the life potential due to malnutrition, pollution, and diseases resulting from the deterioration of the ecosystem and social services. In addition to the deterioration of the social services, the ecosystem contains important threats to this integrity and health. Although it is increasingly accepted that the social system and ecosystem cannot function fully and properly, there are very few attempts for the sustainability of both systems. Within the framework of ethical approaches for assuring sustainability, applied ethics is in the nature of guiding ideas for establishing a sustainable society that will enable people to reintegrate with the ecosystem and human health.

As a general definition, the subject of bioethics is life, not only human life, which is the main subject of medical ethics, but also the life of all organisms that exist in the nature. The subject of environmental ethics is the nature with all its living and

#### *Sustainable Buildings from the Perspective Environmental Bioethics DOI: http://dx.doi.org/10.5772/intechopen.87836*

 nonliving elements. Perhaps the subtle distinction between bioethics and ecocentric ethics, or shortly environmental ethics, can be understood by defining the distinction between life and nature. When we examine the concepts in terms of content, nature has a meaning that also involves life. Nature is a habitat that ensures the continuation of life, nurtures it, protects it, transforms it, and enables it to reproduce itself on its own. A nature without life can exist, whereas a life without nature is out of question. What gives birth to life is nature. At this point, in terms of meaning, it would not be wrong to say that many thoughts are within the bioethical environmental ethics, as also believed by human beings. However, from a practical point of view, life has to recognize and cover all elements of nature, for it cannot sustain itself without preserving nature. A view which places life in the center cannot isolate itself from its nature. In other words, bioethics is a wide area that includes environmental ethics.

The future of the bioenvironment endangered by human actions is an important issue of our time. This issue has also an ethical dimension. The existence of humanity is closely related to and dependent on all life on earth. In that case, how can we reconcile the rapid destruction of life (or bios) with our existence? At this very point, it becomes necessary to shift to the bio-centered, in other words lifecentered, perspective. This transition, as mentioned earlier, has to take place in every aspect of life. People should not put themselves in the heart of every decision they take or every policy they apply, but they must put life, which they are part of themselves, in the center.

Bioethics is a concept that has emerged as a result of such a requirement. Nevertheless, as we cannot think of a life that is without any environment, the environmental bioethics appears as a concept in the intersection of environmental ethics and bioethics. In the solution of the current global ethical problem sets that we encounter during the formation of social policies, we must create a sustainable future with the biopolitics based on the concept of "environmental bioethics."

 In order to create sustainable living spaces, ecological cities, and sustainable buildings of the future, first it is important that the structural elements which constitute the management systems of societies adopt this concept. So as to have an ecological city model, environmental bioethical values and political culture of the urban society must be formed. Lack of environmental bioethical value and inadequate level of organization of democratic elements can be mentioned among the most dominant reasons for the gap between what is known to be best for the environment in human actions and what is actually applied [4–6].

#### **3. Sustainable buildings as an environmental bioethical value**

Sustainable buildings are the product of the biopolitics, which are primarily formed by a holistic environmental bioethical approach. Based on the fact that the idea sets the action, sustainable buildings can be accepted as the practical reflection of environmental bioethics and biopolitics. While trying to find answers to openended questions in humans to human relations, ethics used to make assessments based on "good" and "bad," limiting the responsibility aspect, which is an important component of it, only to human beings.

With the concept of bioethics, the scope of human responsibility has been extended to their relationship with the ecosystem together with all its components such as animals, plants, air, and water. Mankind is part of the nature, not the center of it. But they are responsible against the whole. In each decision taken, bioethics requires that minimum harm be given to the other life forms; the ecological cycles not be broken, to the extent it is possible; and the interference be at the minimum level. The first step is to internalize the respect for other life forms [1–3].

 Reducing the destruction can also be considered within the scope of protection. Sustainable buildings that protect and improve the environment indirectly can thus be considered a bioethical value.

Urban life is the search of a heterogeneous society, which must live together, for ways to live in the best and most harmonious way. What is best is relative. The values generated and shared by urban people also generate the system of values of urban people. Urban social values and individual values are in mutual interaction [5–8].

 As defined in the Cultural Dictionary, "Through values, people become aware of their social relationships, goals and historical roles. With the help of values, they find a place within the community; they form their own interests, norms, life styles, world views, etc. individually. Values act as the active driving force of human actions in the process of social life" [9]. In this case, urban values are the material and mental driving force of the facilities of individuals in the urban life and ultimately of the society to realize themselves toward the best. "The best," which is represented by the values in the ecological city, is expected to include the decisions that respect the ecological balances and life in the world, namely, the practice of environmental bioethics.

 The system of values functions through the transformation of the good behavioral rules within that society into normative rules and supervision of these rules via a system of sanctions, and these values are respected. These two mechanisms, namely, the expectation of roles and sanctions, are adopted by the institutions that ensure the order of the society. If there is an established bioethical point of view in the value system, the society, and the individual, self-sacrificing efforts for the ecological city can be continuously developed [9].

It can be said that such a society that has adopted the idea of environmental bioethics represents the ideal of ecological society. According to Aykut N. Çoban [10], the political project of environmentalism is the establishment of an ecological/sustainable society: However, the problem with transition to such society is the assumption that the material structures such as the economy can be transformed by changing ethics/beliefs, which is an exaggeration of the change of individuals' ideas. Other dynamics which affect the development of cities, such as globalization, economy, and politics, are inevitably important. Nevertheless, there are many good and bad examples in the history that show how ideas are directing societies [8, 10].

Considering sustainable buildings as a purpose; taking all kinds of legal, political, and technical decisions for this purpose; and, consequently, the realization and expansion of the ecological city are closely related with the system of individual and social values. Most of the time, the reason for the gap between knowing and applying the truth is the lack of knowledge of ethical values. Tekeli expresses as follows the necessity for integrating theories of education and democracy: "Every theory of democracy that does not relate the theory of education to itself is incomplete." The desired education is an education nourished by philosophy and art, where especially children and young people will learn how to think, question, and learn [2].

 According to Arvanitis, the aim and responsibility of bio-education is "to revive the spirit of humanity to reverse the established value judgments that lead to significant environmental destruction." Bio-education strengthens the idea of integrating environmental concerns to each human effort and enables us to become aware of the interdisciplinary nature of environmental protection [11].

Especially in the sustainable buildings to be constructed in cities, it should be considered as a bridge of communication between pets, urban people, and other bios. The value that children should gain through pets is to respect the unique worlds and needs of animals [12].

*Sustainable Buildings from the Perspective Environmental Bioethics DOI: http://dx.doi.org/10.5772/intechopen.87836*

 It is important that sustainable buildings be addressed in terms of environmental bioethics, instead of a human-centered perspective. This is critical so that the urban environmental problems can be comprehended deeply and widely and can be overcome rather than achieving superficial successes. Thus, besides the future of human beings, sustainable buildings should be defended as a requirement of environmental bioethical consciousness and responsibility. If a human-centric view is adopted while designing an ecological city constructed with sustainable buildings, even if physical arrangements are realized, it will not be possible to understand and resolve the mental, economic, and political relations which are the source of environmental problems. The difference between preserving other lives by seeing them as means and respecting their essential value in terms of ecological balance is important. It is especially important when it comes to interests, because it will determine our decisions, preferences, and priorities. While taking a decision to destruct a wildlife area, whose material value cannot be measured, for the sake of urban income as a measurable instrumental value, it can simply be sacrificed by attributing a purely instrumental value to it. However, our point of view will differ when we attribute an essential value [6–8, 12–14].

#### **4. Conclusion and evaluation**

 Considering the fact that right to life is valid for all living beings all over the world, all practices that threaten this right can be characterized as being against the bioethical values. Today, the main reason behind the environmental problems is the erosion of global bioethical values. Therefore, the environmental damages caused by today's construction practices concern not only the cities they are in but the whole world. Sustainable buildings, by definition, are compatible with ecosystem, sustainable and self-sufficient in terms of resource utilization, and are compatible with environmental bioethical values.

 Building construction, which has been seen as the source and symbol of civilization throughout the history, is controversial today with the existing risks of destructing the civilization. Today, in the world and in Turkey, the root of urban and environmental problems is based on rapid and unplanned industrialization, the fact of continual profit, and not using the technology for the benefit of humans and nature. In this regard, the cities where the majority of human population resides have turned into places that are alienated to the humans as a result of consumption society, "developments" based on continual profit, and raid and plunder. The most important urban problems of our time, namely, poverty, immigration, housing problem, infrastructural deficiencies, and environmental pollution, are the products of a system, which is based on capital needs, not on human life and needs. An understanding based on environmental bioethical values will ensure sustainability of life.

Migration from rural areas to cities and cheap labor have always been encouraged in line with the needs of capital. Today, the urban living environment in our country is characterized by unplanned construction, illegal housing, and poor urban people. Turkey's urbanization level is found to be 55.4% in 1990. And today, 92.3% of Turkey's population live in cities and towns [15]. Environmental problems have increased to a non-negligible level through urbanization that has evolved mainly by this way. The increase in population and urbanization will also increase urban environmental problems. The ecological city model and sustainable buildings are the products of such a search. The international nature of the capital has become more important in recent years, and the need for capital restructuring with the economic crisis in the 1970s has brought up the capital accumulation and

the dimensions of this accumulation process in terms of social classes and space. Therefore, the city has gained a special importance as the focus of capital accumulation. In order to achieve the goal of transforming the modern cities that emerged in the Industrial Revolution into ecological cities, it should also be aimed to ensure the economic, political, social, and environmental justice. Because just like the rings of the chain in the ecological cycle, these are all interconnected.

It is obvious that the sustainable buildings are a requirement in the world and in Turkey. A system should be developed where renewable energy resources can be used easily in the buildings and the technology which enables that can be mounted to all kinds of buildings. Development of such system is technically possible and is a must in terms of environmental bioethics.

#### **Author details**

Nesrin Çobanoğlu Department of Medical Ethics and History, Gazi University, Ankara, Turkey

\*Address all correspondence to<EMAIL_ADDRESS>
© 2019 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.

*Sustainable Buildings from the Perspective Environmental Bioethics DOI: http://dx.doi.org/10.5772/intechopen.87836*

#### **References**

[1] Çobanoğlu N. Theoretical and Applied Medicine Ethics. Ankara, Turkey: Eflatun Publishing House; 2009. p. 242. (In Turkish)

[2] Carson R. Silent Spring. Boston: The Riverside Press Cambridge; 1962

[3] Jardins JR. Environmental Ethics Introduction to Environmental Philosophy (Translated by Ruşen Keleş). Ankara: İmge Bookstore; 2006. p. 18. (In Turkish)

[4] Jean R. City and Folk. Ankara, Turkey: Ütopya Publications; 1999. p. 9. (In Turkish)

[5] Kuban. Istanbul an Urban History: Byzantion, Constantinopolis. Istanbul: Türkiye İşbankası Kültür Yay; 1996

 [6] Register R. Ecocities: Rebuilding Cities in Balance with Nature. Revised ed. Canada: New Society Publishers; 2006

[7] Tekeli I. Participatory Democracy and Civil Society Organizations. Ankara: Publication of Social Democracy Association; 2004. p. 10. (In Turkish)

[8] Dewey J. Freedom and Cultur, (Translation. Günyol V.). İstanbul: Remzi Publishes; 1987. (In Turkish)

[9] Cultural Dictionary, (Translated and Edited by Aziz Çalışlar), Altın Kitaplar Publishing House; 1983. p. 66. (In Turkish)

[10] Çoban NA. Articulation of Ideological Elements of Environmentalism. Journal of Political Sciences Faculty. 2002;**57**(3):28-30. (In Turkish)

[11] Agni Vilavianos Arvanitis, Biopolitics: Establishing a Vision and Hope Society, Tribute to Rusen Keles Series, Environment and Politics

Longing for Another World (Compiled by Ayşegül Mengi), Imge Bookstore, Ankara; 2007. p. 294. (In Turkish)

[12] Gazi S, Çobanoğlu N. Reflection of Urban Culture Elements in Cities as a Biological Value, Planlama Journal, TMMOB Chamber of City Planners Publication, Ankara; 2008/1-2. p. 19. (In Turkish)

[13] Newman P, Jennings I. Cities as Sustainable Ecosystems. Washington, D.C., USA: Island Press; 2009. p. 15

[14] Bacaksız TS, Çobanoğlu N. Turkey's Biopolitical Policies within the scope of Ecological Buildings, International Sustainable Buildings Symposium (ISBS); 26-28 May 2010; Ankara, Turkey. (In Turkish)

[15] [Internet]. Available from: http://www.tuik.gov.tr/PreTablo. do?alt\_id=1059

**863**

formed:

**Chapter 70**

**Abstract**

design

**1. Introduction**

**2. Transformation**

*Hulki Cevizoğlu*

Urban Empires

Sustainable Urban Design and

In this chapter, structural changes (both building and human structures) in metropolises in our age, which is defined as late modernity, postmodernity, or fluid modernity, are discussed. Discriminatory growth between cities, primate city formations, growth, collapses, depressions, migration, marginalization, freedoms, gentrification studies, security, surveillance and sustainable urban design, and environmental policies are among the main research topics. In addition, a new

**Keywords:** urban empires, modernity, structural change, freedom, sustainable urban

Bauman argues that his sociology is "a way of thinking about human world." Mills places "sociological imagination" at the basis of sociological thought and analysis. When the developmental process of the modern society is taken into consideration, the place of the classics introduces the problematic of "how society

This presentation aims to discuss the realities experienced in the contemporary world with the sociological theories of the classical period by referring to the concepts such as "poverty, hunger, new colonialism, migration, technological changes, globalization, inequalities, polarization of economic sources, and human rights."

The title above does not include Kafka's "*Metamorphosis,*" it rather naturally includes social and urban sociological transformation. The world has evolved from the bipolar world of the "cold war" period to the unipolar world upon the dissolution of the Soviet Union. Later, the electronic and internet revolution has made the world a "global village." The geographical, physical, economic, and electronic borders are shattered. In the expressions of Hardt and Negri, the "*Empire*" has been

The hegemony has achieved a new form, and this form is composed of a series of national and supranational organs united under one type of governing logic. This

type of new global hegemony is the thing we call the empire [2].

concept is introduced to the literature as "Urban Empires."

needs to understand its historical origins" [1].

#### **Chapter 70**


*2.2. Chemical Composition of Mentha EOs*

Selected GC–MS analysis chromatograms of the two studied EOs are shown in Figures 2 and 3. The component relative concentrations (calculated as GC peak area percentages and retention index) are shown in Tables 1 and 2. GC–MS results identified that the principal bioactive components belonged to different chemical groups. The major constituents in *M. spicata* EO included thymol (28.19%), adipic acid (25.82%), piperitone (24.76%), and menthol (24.18%), whereas, the major constituents in *M. longifolia* EO included menthol (51.4%), menthyl acetate (20.5%), and d-limonene (11.15%).

**Figure 2.** GC–MS chromatogram of *M. spicata* EO. Where (\*) is the Total Ion Current.

**Figure 3.** GC–MS chromatogram of *M. longifolia* EO. Where (\*) is the Total Ion Current.

MS: Mass spectrometry (GC–MS).


**Tectonic Process of the Sedimentary Basin Formation and Evolution in the Late Cenozoic Arc-Arc Collision Zone, Central Japan**

Akira Takeuchi

[38] Kano, K. and Yoshida, F., 1984, Radiometric ages of the Neogene in central eastern Shimane Prefecture, Japan and their implication in stratigraphic correlation. Bulletin

[39] Kano, K., Yamauchi, S., Takayasu, K., Matsuura, H., and Bunno, M., 1994, Geology of the Matsue District. With Geological Sheet Map at 1:50, 000, Geological Survey of Ja‐

[40] Van Wagoner, J.C., Posamentier, H.W., Mitchum, R.M., Vail, P.R., Sarg, J.F., Loutit, T.S. and Hardenbol, J., 1988, An overview of the fundamentals of sequence stratigra‐

[41] Baker, B.H., 1986, Tectonics and volcanism of the southern Kenya Rift Valley and its influence on rift sedimentation. Geological Society of London Special Publication, 25,

[42] Saneyoshi, M., Nakayama, K., Sakai, T., Sawada, Y. and Ishida, H., 2006, Half-graben filling processes in the early phase of continental rifting: The Miocene Namurungule

[43] Sawada, Y., Saneyhoshi, M., Nakayama, K., Sakai, T., Itaya, T., Hyodo, M., Mukyo‐ kya, Y., Pickford, M. Senut, B., Tanaka, S., Chujo, T. and Ishida, H., 2006, The ages and geological backgrounds of Miocene Nacholapithecus, Samburupithecus, and Or‐ rorin from Kenya. In: Human Origins and Environmental Backgrounds (Eds H. Ishi‐ da, R.H. Tuttle, M. Pickford, N. Ogihara, and M. Nakatsukasa), Springer, New York,

[44] Sakai, T., Saneyoshi, M., Tanaka, S., Sawada, Y., Nakatsukasa, M., Mbua, E. and Ishi‐ da, H., 2010, Climate shift around 10 Ma recorded in Miocene succession of Samburu Hills, northern Kenya Rift, and its significance. Geological Society of London Special

[45] Kunimatsu, Y., Nakatsukasa, M., Sawada, Y., Sakai, T., Hyodo, M., Hyodo, H., Itaya, T., Nakaya, H., Saegusa, H., Mazurier, A., Saneyoshi, M., Tsujikawa, H., Yamamoto, A. and Mbua E.,2007, A new Late Miocene great ape from Kenya and its implica‐ tions for the origins of African great apes and humans. Proceedings of the National

[46] Gorzáles-Acebrón, R., Arribas, J., Mas, R., 2007, Provenance of fluvial sandstones at the start of late Jurassic-Early Cretaceous in the Cameros Basin (N. Spain). Sedimen‐

[47] Cande, S.C. and Kent, D.V., 1995, Revised calibration of magnetostratigraphic polari‐ ty timescale for the Late Cretaceous and Cenozoic. Journal of. Geophysical Research,

[48] Johnson, C.L. and Graham, S.A., 2004, Cycles in perilacustrine facies of late Mesozoic rift basins, southeastern Mongolia. Journal of Sedimentary Research, 74 786-804.

of Geological Survey of Japan, 35, 159-170.(in Japanese with English Abstract)

pan, 57 p.(in Japanese with English Abstract)

45-57.

71-96.

Publication, 342, 109-127.

tary Geology, 202, 138-157.

100, 6093-6095.

Academy of Sciences, 104, 19220-19225.

phy and key definitions. SEPM Special Publication, 42, 39-45.

108 Mechanism of Sedimentary Basin Formation - Multidisciplinary Approach on Active Plate Margins

Formation of the Kenya Rift. Sedimentary Geology, 186, 111-131.

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/56805


**Surface and Bulk Modification of Synthetic Textiles to Improve Dyeability**

Mazeyar Parvinzadeh Gashti1, Julie Willoughby2 and Pramod Agrawal3,4 *1Department of Chemistry and Biochemistry, University of Bern, Bern 2Fiber and Polymer Science Program, Department of Textiles Engineering, Chemistry and Science, North Carolina State University, Raleigh NC, 3Engineering of Fibrous Smart Materials, Department of Engineering Technology, University of Twente, Enschede, 4Knowledge Centre Design & Technology, School of Applied Art & Technology, Saxion University of Applied Sciences, Enschede, 1Switzerland 2USA 3,4The Netherlands*


2.2.1. Characterisation of ESBL Harbouring Gram Negative Bacilli

Identified Enterobacteriaceae were characterized for their resistance pattern by susceptibility testing according to EUCAST (EUCAST V2.0, 2012) [22], with ampicillin (AM), amoxicillin/clavulanic acid (AMC), piperacillin/tazobactam (TZP), cefalexin (CN), cefuroxime (CXM), cefoxitin (FOX), cefotaxime (CTX), ceftazidime (CAZ), cefepime (FEP), imipenem (IPM), meropenem (MEM), gentamicin (GM), trimethoprim/sulfamethoxazole (SXT), nalidixic acid (NA), ciprofloxacin (CIP), moxifloxacin (MOX), tetracycline (TE) and chloramphenicol (C) BD BBLTM Sensi-DiscTM paper discs (BD, Sparks, MD, USA). The inhibition zone diameters were interpreted according to EUCAST guidelines, except Enterobacteriaceae tested for tetracycline, chloramphenicol and nalidixic acid, which were evaluated by the Clinical Laboratory Standards Institute (CLSI, 2011) guidelines [23]. There are no interpretation guidelines for zone diameters of these three antibiotics according to EUCAST.

*E. coli* 25299 was used as reference. The inhibition zone diameters were interpreted according to EUCAST guidelines. The antimicrobials tested and resistance breakpoints applied can be found in the Supplementary Materials (Table S1).

All isolates were screened for ESBL gene families, *bla*CTX-M-1group, *bla*CTX-M-2group, *bla*CTX-M-9group*, bla*GES*, bla*SHV, *bla*TEM, and *bla*VEB by PCR and sequencing as described previously [24,25]. False-positive (not estimated Enterobacteriaceae) strains growing on ESBL-media with a green or brownish colour were identified as Pseudomonadales and Aeromonadales; we decided to include them in the study and therefore these strains were also screened for ESBL genes. Identified Pseudomonadales were characterized for their resistance pattern by susceptibility testing according to EUCAST (EUCAST V2.0, 2012); piperacilin/tazobactam (TZP), ceftazidime (CAZ), cefepime (FEP), meropenem (MEM), imipenem (IPM), amikacin (AN), gentamicin (GM), tobramycin (NN), ciprofloxacin (CIP) and levofloxacin (LEV).


**Author details**

Vladimir A. Testov Mathematics Department, Vologda State University, Vologda, Russia

\*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.


**Author details**

The type strains of *C. bescii* typically yield a mixture of lactic and acetic acid in addition to hydro-

have been noted to produce low ethanol titers. Work by Cha [89] deleted the gene coding for lactate dehydrogenase by introducing a non-replicating plasmid via marker replacement. The resultant knockout strain did demonstrate increased biomass yield as well as acetate and hydrogen production with no lactate production when grown on cellobiose and lactose as well as switch grass hydrolysates. Subsequent work by Chung [81] inserted a NADHdependent *adhE* gene (from *Clostridium thermocellum*) into the *ldh* mutant (JWCB018) result-

but produced 14.8 mM of ethanol from 29.2 mM cellobiose or 12.7% of the theoretical yield. It should be noted that this strain only used a small portion (4.4 mM of 29.2 mM cellobiose) provided and not produce ethanol above 65°C. Work by Cha [89] and Chung [93] introduced the alcohol dehydrogenase genes (*adhB* and *adhE*) from *Thermoanaerobacter pseudoethanolicus* into the *ldh* deficient strain. The two resultant strains yielded ethanol at temperatures greater than 65°C although titers were lower than the aforementioned strain JWCB032 (*ldh*− *adhE+*

The *C. thermocellum* strain with *adhB* only produced 1.4 mM ethanol on avicel and 0.4 mM on switch grass while a strain with *adhE* gave 2.3 and 1.6 mM of ethanol on avicel and switch grass, respectively. One of the reasons for suggested for the low ethanol titers is the availability of cofactors and it should be noted that *T. pseudoethanolicus* ADHs utilize NADPH while the gene products from *C. thermocellum* use NADH as a source of reducing potential. Additional work is therefore needed to more carefully mimic the complex NAD(P)H system

Overall, efforts to engineer thermophilic anaerobes to increase ethanol titers has resulted in modest gains in yields while minimizing or eliminating the formation of unwanted end products. Future targets for genetic manipulation might include the inclusion of the cellulolytic machinery of *C. thermocellum* into highly ethanologenic *Thermoanaerobacter* and

Bioethanol production from lignocellulosic biomass with thermophilic bacteria needs robust microbes regarding several aspects. One of the main advantages of thermophilic bacteria is their broad substrate spectra with many strains capable of simultaneous pentose and hexose degradations. Additionally, some thermophiles degrade complex carbohydrates like cellulose and hemicellulose although many of these strains are not highly ethanologenic. Recent advantages in genetic engineering have improved ethanol yields, mostly by knocking out pathways of undesired end-products. On the back side is the fact that yields and ethanol tolerance as well as low tolerance for high initial substrate concentrations still limits the use of thermophiles for large scale operations. The use of stable co-cultures where on microbe hydrolyses the sugar polymers and the other one ferments the sugars released to ethanol is an

ing in strain *C. bescii* JWCB032. The resultant *ldh*− *adhE*+

of multiple ADHs in *Thermoanaerobacter pseduoethanolicus*.

attractive way to go forward but warrants further investigations.

*Thermoanaerobacterium* strains.

**7. Conclusions**

as end-products although other strains within the genus of *Caldicellulosiruptor*

strain yielded less acetate (4.3 mM)

).

gen and CO2

112 Fuel Ethanol Production from Sugarcane

Sean Michael Scully and Johann Orlygsson\*

\*Address all correspondence to<EMAIL_ADDRESS>
Faculty of Natural Resource Sciences, University of Akureyri, Akureyri, Iceland

## **References**

[13] Wagner ID, Wiegel J. Diversity of thermophilic anaerobes. In: Incredible anaerobes: From physiology to genomics fuels. Annals of the New York Academy of Sciences. 2008; **1125**:1-43

[24] Euzéby JP. List of bacterial names with standing in nomenclature: A folder available on

Progress in Second Generation Ethanol Production with Thermophilic Bacteria

http://dx.doi.org/10.5772/intechopen.78020

115

[25] Parte AC. LPSN-list of prokaryotic names with standing in nomenclature. Nucleic Acids

[26] Wiegel J, Tanner R, Rainey FA. An introduction to the family clostridae. In: Dworkin M, Falkow S, Rosenberg E, Schleifer K-H, Stackebrandt E, editors. The Prokaryotes, 3rd ed.

[27] Canganella F, Wiegel J. The potential of thermophilic clostridia in biotechnology. In: Woods DR, editor. The Clostridia and Biotechnology. Vol. 23. Freepost, England: Butter-

[28] Carreira LH, Ljungdahl LG. Production of ethanol from biomass using anaerobic thermophilic bacteria. In: Wise DL, editor. Liquid Fuel Developments. Boca Raton, Flordia,

[29] Nölling J, Breton G, Omelchenko MV, Makarova KS, Zeng Q, Gibson R, Lee HM, Dubois J, Qiu D, Hitti J, Wolf YI, Tatusov RL, Sabathe F, Doucette-Stamm L, Soucaille P, Daly MJ, Bennett GN, Koonin EV, Smith DR. Genome sequence and comparative analysis of the solvent-producing bacterium *Clostridium acetobutylicum*. Journal of Bacteriology.

[30] Sabathe F, Belaich A, Soucaille P. Characterization of the cellulolytic complex (cellulosome) of *Clostridium acetobutylicum*. FEMS Microbiology Letters. 2002;**217**:15-22

[31] Han SO, Yukawa H, Inui M, Doi RH. Transcription of *Clostridium cellulovorans* cellulosomal cellulase and hemicellulase genes. Journal of Bacteriology. 2003;**185**:2520-2527

[32] Balusu R, Paduru RMR, Seenyya G, Reddy G. Production of ethanol from cellulosic biomass by *Clostridium thermocellum* SS19 in submerged fermentation: Screening of nutrients using Plackett-Burman design. Applied Biochemistry and Biotechnology. 2004;**117**:133-141

[33] Rani KS, Seenayya G. High ethanol tolerance of new isolates of *Clostridium thermocellum* strains SS21 and SS22. World Journal of Microbiology & Biotechnology. 1999;**15**:173-178

[34] Orlygsson J. Ethanol production from biomass by a moderate thermophile. *Clostridium*

[35] Collins MD, Lawson PA, Willems A, Cordoba JJ, Fernandez-Garayzabal J, Garcia P, Cai J, Hippe H, Farrow JA. The phylogeny of the genus *Clostridium*: Proposal of five new genera and eleven new species combinations. International Journal of Systematic

[36] Schink B, Zeikus JG. *Clostridium thermosulfurogenes* sp. nov, a new thermophile that produces elementar sulfur from thiosulfate. Journal of General Microbiology. 1983;**129**:

[37] Lee YE, Jain MK, Lee C, Lowe SE, Zeikus JG. Taxonomic distinction of saccharolytic thermophilic anaerobes: Description of *Thermoanaerobacterium xylanolyticum* gen. Nov., sp. nov., and *Thermoanaerobacterium saccharolyticum* gen. Nov., sp. nov.; reclassification

the. International Journal of Systematic Bacteriology. 1997;**47**:590-592

Springer: New York, NY, USA; 2006; part 1. pp. 654-678

USA: CRC Press, ISBN<PHONE_NUMBER>; 1993. pp. 1-28

AK1. Icelandic Agricultural Science. 2012;**25**:25-35

Bacteriology. 1994;**44**:812-826

1145-1158

Research. 2014;**42**:D613-D616

worth-Heinemann; 1993. pp. 394-429

2001;**183**:4823-4838

[24] Euzéby JP. List of bacterial names with standing in nomenclature: A folder available on the. International Journal of Systematic Bacteriology. 1997;**47**:590-592

[13] Wagner ID, Wiegel J. Diversity of thermophilic anaerobes. In: Incredible anaerobes: From physiology to genomics fuels. Annals of the New York Academy of Sciences. 2008;

[14] Slobodkin AI, Tourova TP, Kuznetsov BB, Kostrikina NA, Chernyh NA, Bonch-Osmolovskaya EA. *Thermoanaerobacter siderophilus* sp. nov., a novel dis-similatory Fe(III)-reducing, anaerobic, thermophilic bacterium. International Journal of Systematic

[15] Wiegel J, Ljungdahl LG. *Thermoanaerobacter ethanolicus* gen. Nov., spec. Nov., a new, extreme thermophilic, anaerobic bacterium. Archives of Microbiology. 1981;**128**:343-348

[16] Larsen L, Nielsen P, Ahring BK. *Thermoanaerobacter mathranii* sp. nov, an ethanolproducing, extremely thermophilic anaerobic bacterium from a hot spring in Iceland.

[17] Jessen JE, Orlygsson J. Production of ethanol from sugars and lignocellulosic biomass by *Thermoanaerobacter* J1 isolated from a hot spring in Iceland. Journal of Biomedicine and

[18] Fong JCN, Svenson CJ, Nakasugi K, Leong CTC, Bowman JP, Chen B, Glenn DR, Neilan BA, Rogers PL. Isolation and characterization of two novel ethanol-tolerant facultative-anaerobic thermophilic bacteria strains from waste compost. Extremophiles.

[19] Sekiguchi Y, Imachi H, Susilorukmi A, Muramatsu M, Ohashi A, Harada H, Hanada S, Kamagata Y. *Tepidanaerobacter syntrophicus* gen. Nov., sp. nov., an anaerobic, moderately thermophilic, syntrophic alcohol- and lactate-degrading bacterium isolated from thermophilic digested sludges. International Journal of Systematic and Evolutionary

[20] Cayol JL, Ollivier B, Patel BKC, Ravot G, Magot M, Ageron E, Grimont PAD, Garcia JL. Description of *Thermoanaerobacter brockii* subsp. *lactiethylicus* subsp. nov., isolated from a deep subsurface French oil well, a proposal to reclassify *Thermoanaerobacter finnii* as *Thermoanaerobacter brockii* subsp.*finnii* comb. nov., and an emended description of *Thermoanaerobacter brockii*. International Journal of Systematic Bacteriology. 1995;

[21] Cann IK, Stroot PG, Mackie KR, White BA, Mackie RI. Characterization of two novel saccharolytic, anaerobic thermophiles, *Thermoanaerobacterium polysaccharolyticum* sp. nov. and *Thermoanaerobacterium zeae* sp. nov., and emendation of the genus *Thermoanaerobacterium*. International Journal of Systematic and Evolutionary Microbiology. 2001;**51**:

[22] Amend JP, Shock EL. Energetics of overall metabolic reactions of thermophilic and hyperthermophilic *Archaea* and bacteria. FEMS Microbiology Reviews. 2001;**25**:175-243

[23] Sveinsdottir M, Baldursson SRB, Orlygsson J. Ethanol production from monosugars and lignocellulosic biomass by thermophilic bacteria isolated from Icelandic hot springs.

**1125**:1-43

114 Fuel Ethanol Production from Sugarcane

2006;**10**:363-372

**45**:783-789

293-302

Microbiology. 2006;**56**:1621-1629

Icelandic Agricultural Sciences. 2009;**22**:45-58

Bacteriology. 1999;**49**:1471-1478

Archives of Microbiology. 1997;**168**:114-119

Biotechnology. 2012;**186982**. DOI: 10.1155/2012/186982

of *Thermoanaerobium brockii*, *Clostridium thermosulfurogenes*, and *Clostridium thermohydrosulfuricum* E100-69 as *Thermoanaerobacter brockii* comb. nov., *Thermoanaerobacterium thermosulfurigenes* comb. nov., and *Thermoanaerobacter thermohydrosulfuricus* comb. nov., respectively; and transfer of *Clostridium thermohydrosulfuricum* 39E to *Thermoanaerobacter ethanolicus*. International Journal of Systematic Bacteriology. 1993;**43**:41-51

[49] Zambare V, Bhalla A, Muthukumarappan K, Sani RK, Christopher L. Bioprocessing of agricultural waste to ethanol utilizing a cellulolytic extremophile. Extremophiles.

Progress in Second Generation Ethanol Production with Thermophilic Bacteria

[50] Crespo RE, Badshah M, Alvarez MT, Mattiasson B. Ethanol production by continuous fermentation of d-(+)-cellobiose, d-(+)-xylose and sugarcane bagasse hydrolysate using the thermoanaerobe *Caloramator boliviensis*. Bioresource Technology. 2012;

[51] He Q, Lokken PM, Chen S, Zhou J. Characterization of the impact of acetate and lactate on ethanolic fermentation by *Thermoanaerobacter ethanolicus*. Bioresource and Technology.

bial metabolism. International Journal of Hydrogen Energy. 2008;**33**:5122-5130

[53] Hallenbeck PC. Fermentative hydrogen production: Principles, progress and prognosis.

[54] Fardeau ML, Patel BKC, Magot M, Ollivier B. Utilization of serine, leucine, isoleucine and valine by *Thermoanaerobacter brockii* in the presence of thiosulfate or *Methanobacterium* sp

[55] Tomás AF, Karakashev D, Angelidaki I. *Thermoanaerobacter pentosaceus* sp. nov., an anaerobic, extreme thermophilic, high ethanol-yielding bacterium isolated from household waste. International Journal of Systematic and Evolutionary Microbiology. 2012;**63**:2396-2404

[56] Hahn Hagerdahl B, Galbe M, Gorwa-Grauslund MF, Liden G, Zacchi G. Bio-ethanol the fuel of tomorrow from residues today. Trends in Biotechnology. 2006;**24**:549-556

[57] Lovitt RW, Longin R, Zeikus JG. Ethanol production by thermophilic bacteria: Physiological comparison of solvent effects on parent and alcohol-tolerant strains of *Clostridium thermohydrosulfuricum*. Applied and Environmental Microbiology. 1984;**48**:171-177

[58] Wang DIC, Avgerinos GC, Biocic I, Wang SD, Fang HY. Ethanol from cellulosic biomass. Philosophical transactions of the Royal Society of London Series B-Biological Sciences.

[59] Hild HM, Stuckey DC, Leak DJ. Effect of nutrient limitation on product formation during continuous fermentation of xylose with *Thermoanaerobacter ethanolicus* JW200 Fe(7).

[60] Lynd LR, Grethlein HE, Wolkin RH. Fermentation of cellulosic substrates in batch and continuous culture by *Clostridium thermocellum*. Applied and Environemental Micro-

[61] Rani KS, Swamy MV, Seenayya G. Increased ethanol production by metabolic modulation of cellulose fermentation in *Clostridium thermocellum*. Biotechnology Letters. 1997;

Applied Microbiology and Biotechnology. 2003;**60**:679-686

http://dx.doi.org/10.5772/intechopen.78020

117

2011;**15**:611-618

**103**:186-191

2009;**100**:5955-5965

1983;**300**:323-333

**19**:819-823

biology. 1989;**55**:3131-3139

[52] Jones P. Improving fermentative biomass-derived H2

as electron acceptors. Anaerobe. 1997;**3**:405-410

International Journal of Hydrogen Energy. 2009;**34**:7379-7389

[49] Zambare V, Bhalla A, Muthukumarappan K, Sani RK, Christopher L. Bioprocessing of agricultural waste to ethanol utilizing a cellulolytic extremophile. Extremophiles. 2011;**15**:611-618

of *Thermoanaerobium brockii*, *Clostridium thermosulfurogenes*, and *Clostridium thermohydrosulfuricum* E100-69 as *Thermoanaerobacter brockii* comb. nov., *Thermoanaerobacterium thermosulfurigenes* comb. nov., and *Thermoanaerobacter thermohydrosulfuricus* comb. nov., respectively; and transfer of *Clostridium thermohydrosulfuricum* 39E to *Thermoanaerobacter*

*ethanolicus*. International Journal of Systematic Bacteriology. 1993;**43**:41-51

end-product formation. Energy and Fuels. 2012;**26**:4568-4574

Systematic and Evolutionary Microbiology. 2007;**57**:1429-1434

BG1L1. Applied Microbiology and Biotechnology. 2007;**77**:61-68

plex biomass by *Thermoanaerobacter* AK5

116 Fuel Ethanol Production from Sugarcane

and Bioengineering. 2012;**109**:686-694

2007;**2**:364-377

114-119

Bacteriology. 1988;**170**:2809-2815

Bioengineering. 2013;**110**:1574-1582

[38] Brynjarsdottir H, Wawiernia B, Orlygsson J. Ethanol production from sugars and com-

[39] Almarsdottir AR, Sigurbjornsdottir MA, Orlygsson J. Effects of various factors on ethanol yields from lignocellulosic biomass by *Thermoanaerobacterium* AK17. Biotechnology

[40] Lacis LS, Lawford HG. Ethanol-production from xylose by *Thermoanaerobacter ethanolicus*

[41] Lee Y-J, Dashti M, Prange A, Rainey FA, Rohde M, Whitman WB, Wiegel J. *Thermoanaerobacter sulfurigignens* sp. nov., an anaerobic thermophilic bacerium that reduces 1 M thiosulfate to elemental sulfur and tolerates 90 mM sulfite. International Journal of

[42] Georgieva TI, Ahring BK. Evaluation of continuous ethanol fermentation of dilute-acid corn Stover hydrolysate using thermophilic anaerobic bacterium *Thermoanaerobacter*

[43] Georgieva TI, Mikkelsen MJ, Ahring BK. High ethanol tolerance of the thermophilic anaerobic ethanol producer *Thermoanaerobacter* BG1L1. Cent. Europ. Journal of Biology.

[44] Lovitt RW, Shen GJ, Zeikus JG. Ethanol-production by thermophilic bacteria – biochemical basis for ethanol and hydrogen tolerance in *Clostridium thermohydrosulfuricum*. Journal of

[45] Larsen L, Nielsen P. *Thermoanaerobacter mathranii* sp. nov., an ethanol-producing extremely thermophilic bacterium from hot spring in Iceland. Archives of Microbiology. 1997;**168**:

[46] Tomás AF, Karagöz P, Karakashev D, Angelidaki I. Extreme thermophilic ethanol production from rapeseed straw: Using the newly isolated *Thermoanaerobacter pentosaceus* and combining it with *Saccharomyces cerevisiae* in a two-step process. Biotechnology and

[47] Svetlitchnyi VA, Kensch O, Falkenhan DA, Korseska SG, Lippert N, Prinz M, Sassi J, Schickor A, Curvers S. Single-step ethanol production from lignocellulose using novel

[48] Crespo C, Pozzo T, Karlsson EN, Alvarez MP, Mattiasson B. *Caloramator boliviensis* sp. nov., a thermophilic, ethanol-producing bacterium isolated from a hot spring. International Journal of Systematic and Evolutionary Microbiology. 2012;**62**:1679-1686

extremely thermophilic bacteria. Biotechnology for Biofuels. 2013;**6**:31

in batch and continuous culture. Archives of Microbiology. 1988;**150**:48-55

: The effect of electron-scavenging systems on

[62] Singh N, Mathur AS, Tuli DK, Gupta RP, Barrow CJ, Puri M. Cellulosic ethanol production via consolidated bioprocessing by a novel thermophilic anaerobic bacterium isolated from a Himalayan hot spring. Biotechnology for Biofuels. 2017;**10**:73

[74] Shaw AJ, Hogsett DA, Lynd LR. Identification of the [FeFe]-hydrogenase responsible for hydrogen generation in *Thermoanaerobacterium saccharolyticum* and demonstration of increased ethanol yield via hydrogenase knockout. Journal of Bacteriology. 2009;**191**:

Progress in Second Generation Ethanol Production with Thermophilic Bacteria

http://dx.doi.org/10.5772/intechopen.78020

119

[75] Desai SG, Guerinot ML, Lynd LR. Cloning of L-lactate dehydrogenase and elimination of lactic acid production via gene knockout in *Thermoanaerobacterium saccharolyticum* JW/

[76] Tripathi SA, Olson DG, Argyros DA, Miller BB, Barrett TF, Murphy DM, Mccool JD, Warner AK, Rajgarhia VB, Lynd LR, Hogsett DA, Caiazza NC. Development of pyrFbased genetic system for targeted gene deletion in *Clostridium thermocellum* and creation

[77] Biswas R, Prabhu S, Lynd LR, Guss AM. Increase in ethanol yield via elimination of lactate production in an ethanol-tolerant mutant of *Clostridium thermocellum*. PLoS One.

[78] Cripps RE, Eley K, Leak DJ, Rudd B, Taylor M, Todd M, Biakes S, Martin S, Atkinson T. Metabolic engineering of *Geobacillus thermoglucosidasius* for high yields ethanol produc-

[79] Yao S, Mikkelsen MJ. Metabolic engineering to improve ethanol production in *Thermoanaerobacter mathranii*. Applied Microbiology and Biotechnology. 2010;**88**:199-208

[80] Andersen RL, Jensen KM, Mikkelsen MJ. Continuous ethanol fermentation of pretreated lignocellulosic biomasses, waste biomasses, molasses and syrup using the anaerobic, thermophilic bacterium *Thermoanaerobacter italicus* Pentocrobe 411. PLoS One. 2015;**10**:8

[81] Chung D, Cha M, Guss AM, Westpheling J. Direct conversion of plant biomass to ethanol by engineered *Caldicellulosiruptor bescii*. Proceedings of the National Academy of

[82] Shaw AJ, Hogsett DA, Lynd LR. Natural competence in *Thermoanaerobacter* and *Thermoanaerobacterium* species. Applied and Environmental Microbiology. 2010;**76**:4713-4719

[83] Argyros DA, Tripathi SA, Barrett TF, Rogers SR, Feinberg LF, Olson DG, Foden JM, Miller BB, Lynd LR, Hogsett DA, Caiazza NC. High ethanol titers from cellulose by using metabolically engineered thermophilic, anaerobic microbes. Applied and Envi-

[84] Xu L, Tschirner U. Immobilized anaerobic fermentation for bio-fuel production by *Clostridium* co-culture. Bioprocess and Biosystems Engineering. 2014;**37**:1551-1559

[85] Tyurin MV, Lynd LR, Wiegel J. Gene transfer systems for obligately anaerobic thermophilic bacteria. In: Rainey FA, Oren A, editors. Methods in Microbiology. Vol. 35.

London, England: Academic Press Ltd. Elsvier Science Ltd; 2006. pp. 309-330

Sciences of the United States of America. 2014;**111**:8931-8936

of a pta mutant. Applied and Environmental Microbiology. 2010;**76**:6591-6599

SL-YS485. Applied Microbiology and Biotechnology. 2004;**65**:600-605

2014:9. DOI: 10.1371/journal.pone.0086389

tion. Metabolic Engineering. 2009;**11**:398-408

ronmental Microbiology. 2011;**77**:8288-8294

6457-6464

[74] Shaw AJ, Hogsett DA, Lynd LR. Identification of the [FeFe]-hydrogenase responsible for hydrogen generation in *Thermoanaerobacterium saccharolyticum* and demonstration of increased ethanol yield via hydrogenase knockout. Journal of Bacteriology. 2009;**191**: 6457-6464

[62] Singh N, Mathur AS, Tuli DK, Gupta RP, Barrow CJ, Puri M. Cellulosic ethanol production via consolidated bioprocessing by a novel thermophilic anaerobic bacterium

[63] Ahring BK, Licht D, Schmidt AS, Sommer P, Thomsen AB. Production of ethanol from wet oxidised wheat straw by *Thermoanaerobacter mathranii*. Bioresource Technology. 1999;

[64] Avci A, Donmez S. Effect of zinc on ethanol production by two *Thermoanaerobacter*

[65] Georgieva TI, Mikkelsen MJ, Ahring BK. Ethanol production from wet-exploded wheat straw hydrolysate by thermophilic anaerobic bacterium *Thermoanaerobacter* BG1L1 in a continuous immobilized reactor. Applied Biochemistry and Biotechnology. 2008;**145**:

[66] Wiegel J, Carreira LH, Mothershed CP, Puls J. Production of ethanol from bio-polymers by anaerobic, thermophilic, and extreme thermophilic bacteria. II. *Thermoanaerobacter ethanolicus* JW200 and its mutants in batch cultures and resting cell experiments. Biotechnology

[67] Ahring BK, Jensen K, Nielsen P, Bjerre AB, Schmidt AS. Pretreatment of wheat straw and conversion of xylose and xylan to ethanol by thermophilic anaerobic bacteria. Bio-

[68] Rani KS, Swamy MV, Seenayya G. Production of ethanol from various pure and natural cellulosic biomass by *Clostridium thermocellum* strains SS21 and SS22. Process

[69] Lin CW, Wu CH, Tran DT, Shih MC, Li WH, Wu CF. Mixed culture fermentation from lignocellulosic materials using thermophilic lignocellulose-degrading anaerobes.

[70] Lynd LR, Weimer PJ, van Zyl WH, Pretorius LS. Microbial cellulose utilization fundamentals and biotechnology. Microbiology Molecular Biology Reviews. 2002;**66**:506-577

[71] Shao X, Raman B, Zhu M, Mielenz JR, Brown SD, Guss AM, Lynd LR. Mutant selection and phenotypic and genetic characterization of ethanol-tolerant strains of *Clostridium*

[72] Sittijunda S, Tomas AF, Reungsang A, O-Thong S, Angelidaki I. Ethanol production from glucose and xylose by immobiliezed *Thermoanaerobacter pentosaceus* at 70°C in an up-flow anaerobic sludge blanket (UASB) reactor. Bioresource Technology. 2013;**143**:598-607 [73] Shaw AJ, Podkaminer KK, Desai SG, Bardsley JS, Rogers SR, Thorne PG, Hogsett DA, Lynd LR. Metabolic engineering of a thermophilic bacterium to produce ethanol at high yield. Proceedings of the National Academy of Sciences of the United States of America.

*thermocellum*. Applied Microbiology and Biotechnology. 2011;**92**:641-652

strains. Process Biochemistry. 2006;**41**:984-989

and Bioengineering. 1983;**13**:193-205

resource Technology. 1996;**58**:107-113

Process Biochemistry. 2010;**46**:489-493

Biochemistry. 1988;**33**:435-440

2008;**105**:13769-13774

isolated from a Himalayan hot spring. Biotechnology for Biofuels. 2017;**10**:73

**68**:3-9

118 Fuel Ethanol Production from Sugarcane

99-110

[86] Mai V, Lorenz WW, Wiegel J. Transformation of *Thermoanaerobacterium* sp. strain JW/ SL-YS485 with plasmid pIKM1 conferring kanamycin resistance. FEMS Microbiology Letters. 1997;**148**:163-167

**Chapter 7**

**Provisional chapter**

**Potential of Thermotolerant Ethanologenic Yeasts**

**Potential of Thermotolerant Ethanologenic Yeasts**

**in High-Temperature Fermentation**

**in High-Temperature Fermentation**

Tomoyuki Kosaka, Noppon Lertwattanasakul, Nadchanok Rodrussamee, Mochamad Nurcholis, Ngo Thi Phuong Dung, Chansom Keo-Oudone,

Tomoyuki Kosaka, Noppon Lertwattanasakul, Nadchanok Rodrussamee, Mochamad Nurcholis, Ngo Thi Phuong Dung, Chansom Keo-Oudone,

Constantinos Theodoropoulos, Suprayogi, Jaya Mahar Maligan, Savitree Limtong and

Constantinos Theodoropoulos, Suprayogi, Jaya Mahar Maligan, Savitree Limtong and

Additional information is available at the end of the chapter

Additional information is available at the end of the chapter

Masayuki Murata, Peter Götz,

Masayuki Murata, Peter Götz,

http://dx.doi.org/10.5772/intechopen.79144

Mamoru Yamada

**Abstract**

Mamoru Yamada

**Isolated from ASEAN Countries and Their Application**

**Isolated from ASEAN Countries and Their Application**

Thermotolerant ethanologenic yeasts receive attention as alternative bio-ethanol producers to traditionally used yeast, *Saccharomyces cerevisiae*. Their utilization is expected to provide several benefits for bio-ethanol production due to their characteristics and robustness. They have been isolated from a wide variety of environments in a number of ASEAN countries: Thailand, Vietnam, Laos, and Indonesia. One of these yeasts, Kluyveromyces marxianus has been investigated regarding characteristics. Some strains efficiently utilize xylose, which is a main component of the 2nd generation biomass. In addition, the genetic basis of *K. marxianus* has been revealed by genomic sequencing and is exploited for further improvement of the strains by thermal adaptation or gene engineering techniques. Moreover, the glucose repression of *K. marxianus* and its mechanisms has been investigated. Results suggest that *K. marxianus* is an alternative to *S. cerevisiae* in next-generation bio-ethanol production industry. Indeed, we have succeeded to apply *K. marxianus* for bio-ethanol production in a newly developed process, which combines high-temperature fermentation with simultaneous fermentation and distillation under low pressure. This chapter aims to provide valuable information on thermotolerant ethanologenic yeasts and their application, which may direct the economic bioproduction of ethanol and other useful materials in the future.

**Keywords:** thermotolerant yeast, high-temperature fermentation, genomic aspects

© 2016 The Author(s). Licensee InTech. 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.

© 2018 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.

DOI: 10.5772/intechopen.79144

#### **Potential of Thermotolerant Ethanologenic Yeasts Isolated from ASEAN Countries and Their Application in High-Temperature Fermentation Potential of Thermotolerant Ethanologenic Yeasts Isolated from ASEAN Countries and Their Application in High-Temperature Fermentation**

DOI: 10.5772/intechopen.79144

Tomoyuki Kosaka, Noppon Lertwattanasakul, Nadchanok Rodrussamee, Mochamad Nurcholis, Ngo Thi Phuong Dung, Chansom Keo-Oudone, Masayuki Murata, Peter Götz, Constantinos Theodoropoulos, Suprayogi, Jaya Mahar Maligan, Savitree Limtong and Mamoru Yamada Tomoyuki Kosaka, Noppon Lertwattanasakul, Nadchanok Rodrussamee, Mochamad Nurcholis, Ngo Thi Phuong Dung, Chansom Keo-Oudone, Masayuki Murata, Peter Götz, Constantinos Theodoropoulos, Suprayogi, Jaya Mahar Maligan, Savitree Limtong and Mamoru Yamada

Additional information is available at the end of the chapter Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/intechopen.79144

#### **Abstract**

[86] Mai V, Lorenz WW, Wiegel J. Transformation of *Thermoanaerobacterium* sp. strain JW/ SL-YS485 with plasmid pIKM1 conferring kanamycin resistance. FEMS Microbiology

[87] Yao S, Mikkelsen MJ. Identification and overexpression of a bifunctional aldehyde/alcohol dehydrogenase responsible for ethanol production in *Thermoanaerobacter mathranii*.

[88] Van Zyl LJ, Taylor MP, Eley K, Tuffin M, Cowan DA. Engineering pyruvate decarboxylase-mediated ethanol production in the thermophilic host *Geobacillus thermoglucosidasius*.

[89] Cha M, Chung D, Elkins JG, Guss AM, Westpheling J. Metabolic engineering of *Caldicellulosiruptor bescii* yields increased hydrogen production from lignocellulosic biomass.

[90] Chung D, Cha M, Farkas J, Westpeling J. Construction of a stable replicating shuttle vector for *Caldicellulosiruptor* species: Use of extenging genetic methodologies to other

[91] Chung D, Farkas J, Westpheling J.Overcoming restriction as a barrier to DNA transformation in *Caldicellulosiruptor* species results in efficient marker replacement. Biotechnology

[92] Chung D, Cha M, Snyder EN, Elkins JG, Guss AM, Westpheling J. Cellulosic ethanol production via consolidated bioprocessing at 75°C by engineered *Caldicellulosiruptor*

[93] Chung D, Verbeke TJ, Cross KL, Wespheling J, Elkins JG. Expression of heat-stable NADPH-dependent alcohol dehydrogenase in *Caldicellulosiruptor bescii* result in furan

aldehyde detoxification. Biotechnology for Biofuels. 2015b;**102**

Journal of Molecular Microbiology and Biotechnology. 2010;**19**:123-133

Applied Microbiology and Biotechnology. 2014;**98**:1247-1259

Letters. 1997;**148**:163-167

120 Fuel Ethanol Production from Sugarcane

Biotechnology for Biofuels. 2013;**6**:85

for Biofuels. 2013b;**6**:82

members of this genus. PLoS One. 2013a;**8**:1-10

*bescii*. Biotechnology for Biofuels. 2015a;**8**

Thermotolerant ethanologenic yeasts receive attention as alternative bio-ethanol producers to traditionally used yeast, *Saccharomyces cerevisiae*. Their utilization is expected to provide several benefits for bio-ethanol production due to their characteristics and robustness. They have been isolated from a wide variety of environments in a number of ASEAN countries: Thailand, Vietnam, Laos, and Indonesia. One of these yeasts, Kluyveromyces marxianus has been investigated regarding characteristics. Some strains efficiently utilize xylose, which is a main component of the 2nd generation biomass. In addition, the genetic basis of *K. marxianus* has been revealed by genomic sequencing and is exploited for further improvement of the strains by thermal adaptation or gene engineering techniques. Moreover, the glucose repression of *K. marxianus* and its mechanisms has been investigated. Results suggest that *K. marxianus* is an alternative to *S. cerevisiae* in next-generation bio-ethanol production industry. Indeed, we have succeeded to apply *K. marxianus* for bio-ethanol production in a newly developed process, which combines high-temperature fermentation with simultaneous fermentation and distillation under low pressure. This chapter aims to provide valuable information on thermotolerant ethanologenic yeasts and their application, which may direct the economic bioproduction of ethanol and other useful materials in the future.

**Keywords:** thermotolerant yeast, high-temperature fermentation, genomic aspects

© 2016 The Author(s). Licensee InTech. 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. © 2018 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.

### **1. Introduction**

Worldwide economic growth with the related increase in CO2 emissions from fossil fuels causes global warming. Utilization of renewable energy with low CO2 emission therefore has been getting increased attention. Renewable energy is generated from renewable natural resources, such as sunlight, wind, rain, tides, waves, geothermal heat, as well as biomass. One such important source of renewable energy, *bio-ethanol*, has been highlighted due to the characteristics of its production from biomass, which is generated by plants using sunlight for CO2 fixation, resulting in carbon neutrality. Bio-ethanol is the name for ethanol produced from biomass by fermentation. This bio-process is thoroughly researched and well-established, and to-date, it produces the most prominent and cost-effective biofuel [1]. Although bio-ethanol production is increasing worldwide and the production of biofuels including ethanol in 2022 is forecasted to be more than 126 billion L [2], biofuels are still more costly than fossil fuels [3]. Therefore, several industrial companies and researchers aim to develop new technologies, enabling the cost-effective production of bio-ethanol from biomass. Since microorganisms are essential for material production through bio-processing, their characteristics and traits are crucial for the production process efficiency. Ethanologenic yeast, *Saccharomyces cerevisiae*, has been traditionally and widely utilized for the production of alcoholic beverages and bioethanol [4, 5]. Industrially common problems in bio-ethanol production related to *S. cerevisiae* strains are temperature level (35–45°C) and high ethanol concentration (over 20%) [6]. These two factors inhibit yeast proliferation and fermentation activity if they reach the upper limit. In addition, for cost-effective bio-ethanol production, the production source must be changed from 1st generation biomass (sugarcane, corn, and wheat, which are important food sources) to 2nd generation biomass (lignocellulosic biomass or woody crops, which are agricultural residues or waste) [7]. Lignocellulosic biomass is composed of hemicellulose, cellulose, and lignin, and the first consists of six (e.g., glucose) and five (e.g., xylose) carbon sugars. However, the low efficiency of ethanol production by *S. cerevisiae* from lignocellulosic biomass hydrolyzates is mainly due to its little ethanol productivity from xylose [8]. Although the *S. cerevisiae* genome encodes all components necessary for xylose utilization, most of them are rarely expressed [9]. In addition, *S. cerevisiae* preferably utilizes glucose while repressing the uptake and catabolism of alternate carbon sources by a mechanism such as glucose repression [10]. This results in the reduction of ethanol production rates from several kinds of biomass. For economically feasible bioethanol production from lignocellulosic biomass, the efficient co-fermentation of glucose and other sugars is also necessary. Therefore, genetic engineering of *S. cerevisiae* strains has been extensively performed, and metabolically engineered strains were developed [11], which have showed higher stress tolerance and/or improved xylose utilization [12, 13]. However, the utilization of genetically recombinant strains in industry has been very limited, especially due to the instability of the desirable phenotype and the necessary confinement to a closed system to prevent their leakage into the environment, which can eventually endanger public health or biodiversity. Therefore, the development of new feasible strains for next-generation bio-ethanol production is under way, and new yeast strains have been isolated that may have advantages compared to *S. cerevisiae*.

belonging to the same genus or even to the same species [14]. These thermotolerant mesophiles are mainly and widely distributed in foods, plants, soils, and waters from tropical environments in ASEAN countries [15]. In these environments, relatively high temperature presumably becomes a selective pressure to enrich thermotolerant strains. These thermotolerant strains are expected to provide a benefit for the industries because they are more robust and resistant to many stressors [14]. In addition, some of these thermotolerant microorganisms can produce distinctive enzymes that function under relatively high temperature conditions [16– 18]. Thermotolerant yeasts have been found and isolated from a number of countries [19–28]. Of these, *K. marxianus* is a haploid, homothallic, thermotolerant, and hemiascomycetous yeast [29, 30]. One such yeast, *K. marxianus* DMKU 3-1042 isolated in Thailand, shows relatively high ethanol productivity and fermentation ability at high temperatures [31], assimilates various sugars including xylose and/or arabinose [32], and exhibits relatively weak glucose repression on utilization of some sugars including sucrose [33]. Therefore, *K. marxianus* is, in comparison to *S. cerevisiae,* a promising candidate for next-generation bio-ethanol production. In addition, the genomic sequences of *K. marxianus* are available [34, 35], and genetic technology and tools have also been developed [36]. Moreover, *K. marxianus* has been a platform for next-generation protein production for structural and biochemical studies [18, 29]. However, it is possible that unidentified and more beneficial thermotolerant yeasts exist in ASEAN countries, especially, thermotolerant high xylose-utilizing and ethanol-producing yeasts, which are needed for 2nd generation biomass utilization. None of the isolated *K. marxianus* strains, however, are able to more efficiently convert xylose to ethanol than strains of

Potential of Thermotolerant Ethanologenic Yeasts Isolated from ASEAN Countries…

http://dx.doi.org/10.5772/intechopen.79144

123

other xylose-utilizing yeasts, such as *Pichia stipitis* (*Scheffersomyces stipitis*) [32, 37].

as thermotolerant strains, leads to lowest energy consumption and emission of CO2

**2. Various ethanologenic thermotolerant yeasts and their**

production [41].

**characteristics**

for ethanol production are presented.

Thermotolerant strains allow the development of high-temperature fermentation (HTF) technology, which enables fermentation at 5–10°C higher than the traditional fermentative process [38, 39]. HTF is thus expected to reduce cooling costs, running costs at the simultaneous saccharification and fermentation (SSF) stage, and contamination risks [6, 31, 38–40], therefore offering a promising technology for bio-ethanol production. Moreover, thermotolerant yeast can also be applied for temperature-uncontrolled fermentation, hence offering another economical advantage. A combination of efficient bioreactors and robust hosts, such

In this chapter, we outline a number of thermotolerant yeasts including *K. marxianus* species isolated in Thailand and their characteristics, including utilization of various sugars, glucose repression, and genetic information, that are beneficial for high-temperature fermentation. In addition, new strains of thermotolerant yeasts that have been isolated in Indonesia, Vietnam, and Laos are summarized. Subsequently, the trial results of HTF with some of these strains

Increasing global energy demand that exceeds the finite supply of fossil fuel has spurred scientific research to deliver alternative fuels. Microbial fermentation and efficient conversion

in biofuel

Recently, thermotolerant microorganisms were found among mesophiles with optimum growth temperatures that are 5–10°C higher than those of the typical mesophilic strains belonging to the same genus or even to the same species [14]. These thermotolerant mesophiles are mainly and widely distributed in foods, plants, soils, and waters from tropical environments in ASEAN countries [15]. In these environments, relatively high temperature presumably becomes a selective pressure to enrich thermotolerant strains. These thermotolerant strains are expected to provide a benefit for the industries because they are more robust and resistant to many stressors [14]. In addition, some of these thermotolerant microorganisms can produce distinctive enzymes that function under relatively high temperature conditions [16– 18]. Thermotolerant yeasts have been found and isolated from a number of countries [19–28]. Of these, *K. marxianus* is a haploid, homothallic, thermotolerant, and hemiascomycetous yeast [29, 30]. One such yeast, *K. marxianus* DMKU 3-1042 isolated in Thailand, shows relatively high ethanol productivity and fermentation ability at high temperatures [31], assimilates various sugars including xylose and/or arabinose [32], and exhibits relatively weak glucose repression on utilization of some sugars including sucrose [33]. Therefore, *K. marxianus* is, in comparison to *S. cerevisiae,* a promising candidate for next-generation bio-ethanol production. In addition, the genomic sequences of *K. marxianus* are available [34, 35], and genetic technology and tools have also been developed [36]. Moreover, *K. marxianus* has been a platform for next-generation protein production for structural and biochemical studies [18, 29]. However, it is possible that unidentified and more beneficial thermotolerant yeasts exist in ASEAN countries, especially, thermotolerant high xylose-utilizing and ethanol-producing yeasts, which are needed for 2nd generation biomass utilization. None of the isolated *K. marxianus* strains, however, are able to more efficiently convert xylose to ethanol than strains of other xylose-utilizing yeasts, such as *Pichia stipitis* (*Scheffersomyces stipitis*) [32, 37].

**1. Introduction**

122 Fuel Ethanol Production from Sugarcane

Worldwide economic growth with the related increase in CO2

causes global warming. Utilization of renewable energy with low CO2

strains have been isolated that may have advantages compared to *S. cerevisiae*.

Recently, thermotolerant microorganisms were found among mesophiles with optimum growth temperatures that are 5–10°C higher than those of the typical mesophilic strains

has been getting increased attention. Renewable energy is generated from renewable natural resources, such as sunlight, wind, rain, tides, waves, geothermal heat, as well as biomass. One such important source of renewable energy, *bio-ethanol*, has been highlighted due to the characteristics of its production from biomass, which is generated by plants using sunlight for CO2 fixation, resulting in carbon neutrality. Bio-ethanol is the name for ethanol produced from biomass by fermentation. This bio-process is thoroughly researched and well-established, and to-date, it produces the most prominent and cost-effective biofuel [1]. Although bio-ethanol production is increasing worldwide and the production of biofuels including ethanol in 2022 is forecasted to be more than 126 billion L [2], biofuels are still more costly than fossil fuels [3]. Therefore, several industrial companies and researchers aim to develop new technologies, enabling the cost-effective production of bio-ethanol from biomass. Since microorganisms are essential for material production through bio-processing, their characteristics and traits are crucial for the production process efficiency. Ethanologenic yeast, *Saccharomyces cerevisiae*, has been traditionally and widely utilized for the production of alcoholic beverages and bioethanol [4, 5]. Industrially common problems in bio-ethanol production related to *S. cerevisiae* strains are temperature level (35–45°C) and high ethanol concentration (over 20%) [6]. These two factors inhibit yeast proliferation and fermentation activity if they reach the upper limit. In addition, for cost-effective bio-ethanol production, the production source must be changed from 1st generation biomass (sugarcane, corn, and wheat, which are important food sources) to 2nd generation biomass (lignocellulosic biomass or woody crops, which are agricultural residues or waste) [7]. Lignocellulosic biomass is composed of hemicellulose, cellulose, and lignin, and the first consists of six (e.g., glucose) and five (e.g., xylose) carbon sugars. However, the low efficiency of ethanol production by *S. cerevisiae* from lignocellulosic biomass hydrolyzates is mainly due to its little ethanol productivity from xylose [8]. Although the *S. cerevisiae* genome encodes all components necessary for xylose utilization, most of them are rarely expressed [9]. In addition, *S. cerevisiae* preferably utilizes glucose while repressing the uptake and catabolism of alternate carbon sources by a mechanism such as glucose repression [10]. This results in the reduction of ethanol production rates from several kinds of biomass. For economically feasible bioethanol production from lignocellulosic biomass, the efficient co-fermentation of glucose and other sugars is also necessary. Therefore, genetic engineering of *S. cerevisiae* strains has been extensively performed, and metabolically engineered strains were developed [11], which have showed higher stress tolerance and/or improved xylose utilization [12, 13]. However, the utilization of genetically recombinant strains in industry has been very limited, especially due to the instability of the desirable phenotype and the necessary confinement to a closed system to prevent their leakage into the environment, which can eventually endanger public health or biodiversity. Therefore, the development of new feasible strains for next-generation bio-ethanol production is under way, and new yeast

emissions from fossil fuels

emission therefore

Thermotolerant strains allow the development of high-temperature fermentation (HTF) technology, which enables fermentation at 5–10°C higher than the traditional fermentative process [38, 39]. HTF is thus expected to reduce cooling costs, running costs at the simultaneous saccharification and fermentation (SSF) stage, and contamination risks [6, 31, 38–40], therefore offering a promising technology for bio-ethanol production. Moreover, thermotolerant yeast can also be applied for temperature-uncontrolled fermentation, hence offering another economical advantage. A combination of efficient bioreactors and robust hosts, such as thermotolerant strains, leads to lowest energy consumption and emission of CO2 in biofuel production [41].

In this chapter, we outline a number of thermotolerant yeasts including *K. marxianus* species isolated in Thailand and their characteristics, including utilization of various sugars, glucose repression, and genetic information, that are beneficial for high-temperature fermentation. In addition, new strains of thermotolerant yeasts that have been isolated in Indonesia, Vietnam, and Laos are summarized. Subsequently, the trial results of HTF with some of these strains for ethanol production are presented.