**Figure 1.** The structures of diapolycopenedioc acids A (**1**), B (**2**) and C (**3**).

*2.2. Methyl 5-Glucosyl-5,6-Dihydro-Apo-4,4ȝ-Lycopenoate from Planococcus Maritimus [29]*

A yellow pigment-producing bacterium (strain iso-3), which was found to be solvent-tolerant, was isolated as an orange-pigmented colony from the microbial analysis of a sample derived from an intertidal sediment from the Clyde estuary, UK. The 16S rRNA gene sequence of strain iso-3 was the most similar to that of type strain *Planococcus maritimus* (99.5 as a similarity score, and 96.4 as an s\_ab score, from the Sequence match analysis of RDP), which belongs to the class Bacilli, phylum Firmicutes, and was identified as *Planococcus maritimus* strain iso-3.

Strain iso-3 was cultured in 100 mL of medium (Marine Broth 2216, Difco, Sparks, MD, USA) in a

500 mL Erlenmeyer flask at 30 °C on a rotary shaker at 120 rpm for 1 day, and the carotenoid produced was purified from the alkaline-digested cells using chromatographic methods (EtOAc/H2O partition ė silica gel column chromatography (CH2Cl2–MeOH (10:1) ė preparative silica gel

HPLC CH2Cl2–MeOH (10:1) ė preparative ODS HPLC (96% MeOH)). A total of 2.5 mg of pure methyl 5-glucosyl-5,6-dihydro-apo-4,4ȝ-lycopenoate (**4**) was obtained from the cells in the 18-liter culture, and the structure of compound **4** was determined by HRESI-MS and spectroscopic (UV-Vis, NMR (1D and 2D investigations on 1H and 13C nuclei), and [΅]D) analyses, as shown in Figure 2.

Compound **4** was a new carotenoid. Compound **4** possessed 5,6-dihydro-5-hydroxy-

apo-4,

4ȝ-lycopene-4ȝ-oic acid (C30 carotenoid) as its aglycone. Although 4,4ȝ-diapocarotene-4-oic acid [32] was previously reported to be a related C30 carotenoid aglycone, 5,6- dihydro and 5-hydroxy functions in the aglycone of compound **4** were demonstrated for the first time. The antioxidant activity

of compound **4** was evaluated using 1O2 suppression activity, and its IC50 value was 5.1 ΐM. We previously described the isolated carotenoid as methyl glucosyl-3,4-dihydroapo-8ȝ-lycopenoate [29], but confirmed its structure as methyl 5-glucosyl-5,6- dihydro-apo-4,4ȝ-lycopenoate, as shown in this review. Corrigenda is currently being prepared for the previous study.

> **Figure 2.** The structure of methyl 5-glucosyl-5,6-dihydro-apo-4,4ȝlycopenoate (**4**).

*2.3. (3R)-Saproxanthin and (3R,2ȝS)-Myxol [30]*

Strain 04OKA-13-27 (MBIC08261) was isolated from the dense mats of filamentous algae from within the territory of damselfish (*Stegastes nigricans*). Strain YM6-073 (MBIC06409) was isolated from a sediment sample collected 0.1 m below the surface of the sea by cultivating for 30 days on an HSV medium. The two marine bacteria, which had been collected off the coast of Okinawa Prefecture, were classified on the basis of this 16S rRNA gene sequences. A similarity search in the databases

of the DNA Data Bank of Japan (DDBJ) and RNA Database Project II (RDPII) showed the 16S rRNA gene sequences of the both strains (04OKA-13-27 and YM6-073) to be 96.5% (1408 bp/1459 bp) similar to *Stanierella latercula* ATCC 23177T, 95.5% (1324 bp/1386 bp) similar to *Gaetbulimicrobium brevivitae* strain SMK-19T, and 94.2% (1306 bp/1386 bp) similar to *Robiginitalea biformata* strain HTCC2501T. The phylogenetic relationship between these strains was deduced with already known species in the family Flavobacteriaceae. The result obtained revealed that the two bacterial strains should be classified as novel species of the family Flavobacteriaceae.

Both 04OKA-13-27 and YM6-073 were cultured in 100 mL of medium (Marine Broth 2216, Difco) in a 500 mL Sakaguchi flask at 30 °C on a rotary shaker at 100 rpm for 1 day, and the carotenoids produced were each purified from the cells using chromatographic methods (EtOAc/H2O partition ė silica gel column chromatography hexane–ethyl acetate (2:1) ė preparative silica gel high performance thin layer chromatography (HPTLC; Merck, Darmstadt, Germany)