Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Sapporo, Japan.
Appl Environ Microbiol. 2014 Feb;80(3):1126-31. doi: 10.1128/AEM.03056-13. Epub 2013 Dec 2.
Medium- and long-chain fatty acids are present in organisms in esterified forms that serve as cell membrane constituents and storage compounds. A large number of organisms are known to accumulate lipophilic materials as a source of energy and carbon. We found a bacterium, designated GK12, that intrinsically accumulates free fatty acids (FFAs) as intracellular droplets without exhibiting cytotoxicity. GK12 is an obligatory anaerobic, mesophilic lactic acid bacterium that was isolated from a methanogenic reactor. Phylogenetic analysis based on 16S rRNA gene sequences showed that GK12 is affiliated with the family Erysipelotrichaceae in the phylum Firmicutes but is distantly related to type species in this family (less than 92% similarity in 16S rRNA gene sequence). Saturated fatty acids with carbon chain lengths of 14, 16, 18, and 20 were produced from glucose under stress conditions, including higher-than-optimum temperatures and the presence of organic solvents that affect cell membrane integrity. FFAs were produced at levels corresponding to up to 25% (wt/wt) of the dry cell mass. Our data suggest that FFA accumulation is a result of an imbalance between excess membrane fatty acid biosynthesis due to homeoviscous adaptation and limited β-oxidation activity due to anaerobic growth involving lactic acid fermentation. FFA droplets were not further utilized as an energy and carbon source, even under conditions of starvation. A naturally occurring bacterium that accumulates significant amounts of long-chain FFAs with noncytotoxicity would provide useful strategies for microbial biodiesel production.
中链和长链脂肪酸以酯化形式存在于生物体中,作为细胞膜成分和储存化合物。已知大量生物体积累亲脂性物质作为能量和碳源。我们发现了一种细菌,命名为 GK12,它内在地积累游离脂肪酸 (FFA) 作为细胞内液滴,而没有表现出细胞毒性。GK12 是一种必需的厌氧、嗜温乳酸细菌,从产甲烷反应器中分离出来。基于 16S rRNA 基因序列的系统发育分析表明,GK12 属于厚壁菌门的埃希氏菌科,但与该科的模式种关系较远(16S rRNA 基因序列相似性小于 92%)。在应激条件下,包括高于最佳温度和影响细胞膜完整性的有机溶剂存在下,从葡萄糖中产生具有 14、16、18 和 20 个碳原子链长的饱和脂肪酸。FFA 的产量可达干细胞质量的 25%(wt/wt)。我们的数据表明,FFA 积累是由于同源粘性适应导致膜脂肪酸生物合成过剩与由于涉及乳酸发酵的厌氧生长导致的有限β-氧化活性之间的不平衡所致。即使在饥饿条件下,FFA 液滴也不会进一步被用作能量和碳源。一种自然积累大量长链 FFA 且无细胞毒性的细菌将为微生物生物柴油生产提供有用的策略。