多轮次胁迫驯化及多因子复合筛选丁醇高产菌

Multi-repeat stress acclimation and butanol high-yielding strain screening by multi-factor complex medium

摘要:

【目的】从陕西省石泉县玉米地土壤中分离获得一株产丁醇菌株并提高其丁醇耐受性和丁醇产量。【方法】采用自行设计的多因子复合筛选方法和丁醇胁迫驯化处理, 在获得丁醇高产菌株的同时提高菌株的丁醇耐受性。【结果】野生菌株D64经多轮次丁醇胁迫驯化处理和多因子复合筛选, 分离获得突变株T64, 其丁醇耐受性明显提高, 能在丁醇浓度为20 g/L的复合筛选培养基上正常生长, 发酵7%玉米醪丁醇产量由13.35 g/L提高到15.18 g/L, 总溶剂(丙酮、丁醇、乙醇)达到21.8 g/L。【结论】采用长时间且丁醇浓度呈梯度渐进增加的胁迫驯化方式, 可使菌种在丁醇的环境中不断进化并有效地提高菌株对丁醇的耐受性。多因子复合筛选方法较其他单一因子筛选方法更为有效, 能较快获得丁醇高产菌。

Abstract:

[Objective] The purpose of this study was to isolate a butanol-producing strain from the soil of corn field in Shiquan County, Shaanxi Province, China, and to improve its butanol tolerance and butanol yield. [Methods] Using the multi-factor complex screening and the treatment of butanol stress acclimation, the strain with a high yield and butanol tolerance was obtained. [Results] The results showed that through several batches of acclimation and screening, the mutant T64 was derived from isolated wild-type strain D64, its butanol tolerance was significantly improved and could grow well in the complex screening medium containing 20 g butanol/L. It produced 21.8 g/L total solvent (acetone, butanol, ethanol) and a butanol yield reached 15.18 g/L using 7% corn mash as fermentation medium, which was higher than that of the wild-type strain D64 (13.35 g/L). [Conclusion] In conclusion, the designed multi-factor complex screening is more effective than the mono-factor screening for obtaining high-producing butanol strain. In addition, the stress acclimation of increasing butanol concentration in long term provides new ideas to research the tolerance of butanol.