Industrial Biotechnology Program, Institute of Chemical and Engineering Sciences, Agency for Science, Technology and Research (A*STAR), 1 Pesek Road, Jurong Island, Singapore 627833.
Appl Biochem Biotechnol. 2012 Nov;168(6):1672-80. doi: 10.1007/s12010-012-9888-0. Epub 2012 Sep 8.
Due to the complex mechanisms involved in butanol-induced stress response, butanol tolerance phenotype is difficult to engineer even in microorganisms with well-defined genetic backgrounds. We therefore aimed to isolate butanol-tolerant microorganisms from environmental samples as potential alternative hosts for butanol production. Soil samples collected were subjected to butanol stress. A microbial strain capable of 2.5-3 % (w/v) butanol tolerance was isolated and identified as Enterococcus faecium by 16S rDNA analysis. The isolate grew readily under both aerobic and anaerobic conditions and was capable of producing butanol anaerobically. In comparison with the obligate anaerobe Clostridium acetobutylicum, the growth under both aerobic and anaerobic conditions of the isolated strain, together with no detection of butyrate and lack of two-phase fermentation suggests different metabolic networks from the obligate anaerobe C. acetobutylicum. Under anaerobic condition, butanol reached up to 0.4 gl(-1) in a batch culture without heterologous introduction of butanol biosynthetic pathway. Besides butanol tolerance, the isolated E. faecium IB1 showed high tolerance to 10 % (w/v) ethanol and 3 % (w/v) isobutanol. With distinct features including high butanol tolerance and natural butanol production, the isolated E. faecium IB1 with minimum metabolic engineering can be explored as a potential host for butanol production.
由于正丁醇诱导应激反应涉及的复杂机制,即使在遗传背景明确的微生物中,也很难对正丁醇耐受性表型进行工程改造。因此,我们旨在从环境样本中分离出具有正丁醇耐受性的微生物,作为潜在的替代生产正丁醇的宿主。采集的土壤样本经正丁醇胁迫处理。通过 16S rDNA 分析,分离并鉴定出一株能耐受 2.5-3%(w/v)正丁醇的微生物菌株,鉴定为屎肠球菌。该分离株在好氧和厌氧条件下均能迅速生长,并能在厌氧条件下产生正丁醇。与专性厌氧菌丙酮丁醇梭菌相比,该分离株在好氧和厌氧条件下的生长情况,以及丁酸的检测不出和无两相发酵表明,其代谢网络与专性厌氧菌丙酮丁醇梭菌不同。在厌氧条件下,在不引入正丁醇生物合成途径的异源情况下,分批培养中可达到 0.4 gl(-1)的正丁醇。除了正丁醇耐受性外,分离得到的屎肠球菌 IB1 对 10%(w/v)乙醇和 3%(w/v)异丁醇也有很高的耐受性。该分离得到的屎肠球菌 IB1 具有高正丁醇耐受性和天然正丁醇生产能力,具有代谢工程最小化的特点,可作为生产正丁醇的潜在宿主进行探索。