Sakurama Haruko, Kishino Shigenobu, Mihara Kousuke, Ando Akinori, Kita Keiko, Takahashi Satomi, Shimizu Sakayu, Ogawa Jun
Laboratory of Industrial Microbiology, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
Laboratory of Industrial Microbiology, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan Laboratory of Fermentation Physiology and Applied Microbiology, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.
J Lipid Res. 2014 Sep;55(9):1855-63. doi: 10.1194/jlr.M045450. Epub 2014 Jul 7.
The PUFAs include many bioactive lipids. The microbial metabolism of C18 PUFAs is known to produce their bioactive isomers, such as conjugated FAs and hydroxy FAs, but there is little information on that of C20 PUFAs. In this study, we aimed to obtain anaerobic bacteria with the ability to produce novel PUFAs from C20 PUFAs. Through the screening of ∼100 strains of anaerobic bacteria, Clostridium bifermentans JCM 1386 was selected as a strain with the ability to saturate PUFAs during anaerobic cultivation. This strain converted arachidonic acid (cis-5,cis-8,cis-11,cis-14-eicosatetraenoic acid) and EPA (cis-5,cis-8,cis-11,cis-14,cis-17-EPA) into cis-5,cis-8,trans-13-eicosatrienoic acid and cis-5,cis-8,trans-13,cis-17-eicosatetraenoic acid, giving yields of 57% and 67% against the added PUFAs, respectively. This is the first report of the isolation of a bacterium transforming C20 PUFAs into corresponding non-methylene-interrupted FAs. We further investigated the substrate specificity of the biohydrogenation by this strain and revealed that it can convert two cis double bonds at the ω6 and ω9 positions in various C18 and C20 PUFAs into a trans double bond at the ω7 position. This study should serve to open up the development of novel potentially bioactive PUFAs.
多不饱和脂肪酸(PUFAs)包括许多生物活性脂质。已知C18多不饱和脂肪酸的微生物代谢会产生其生物活性异构体,如共轭脂肪酸(FAs)和羟基脂肪酸,但关于C20多不饱和脂肪酸的微生物代谢信息却很少。在本研究中,我们旨在获得能够从C20多不饱和脂肪酸产生新型多不饱和脂肪酸的厌氧细菌。通过对约100株厌氧细菌的筛选,双发酵梭菌JCM 1386被选为在厌氧培养过程中具有使多不饱和脂肪酸饱和能力的菌株。该菌株将花生四烯酸(顺式-5,顺式-8,顺式-11,顺式-14-二十碳四烯酸)和二十碳五烯酸(EPA,顺式-5,顺式-8,顺式-11,顺式-14,顺式-17-EPA)转化为顺式-5,顺式-8,反式-13-二十碳三烯酸和顺式-5,顺式-8,反式-13,顺式-17-二十碳四烯酸,相对于添加的多不饱和脂肪酸,产率分别为57%和67%。这是关于分离出一种将C20多不饱和脂肪酸转化为相应非亚甲基中断脂肪酸的细菌的首次报道。我们进一步研究了该菌株生物氢化的底物特异性,发现它可以将各种C18和C20多不饱和脂肪酸中ω6和ω9位置的两个顺式双键转化为ω7位置的一个反式双键。本研究应有助于开启新型潜在生物活性多不饱和脂肪酸的开发。