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诱变育种与基因改造相结合的方法提高了油井本土铜绿假单胞菌的鼠李糖脂产量。

Combined methods of mutation breeding and genetice modifications enhanced rhamnolipids production of oil well Indigenous Pseusdomonas aeruginosa.

作者信息

Liu Mingxin, Sun Jin, Song Xin

机构信息

State Key Laboratory of Microbial Technology, Shandong University, Qingdao, 266237, Shandong Province, China.

National Glycoengineering Research Center, Shandong University, Qingdao, 266237, Shandong Province, China.

出版信息

Sci Rep. 2025 Jun 4;15(1):19673. doi: 10.1038/s41598-025-04671-4.

Abstract

Rhamnolipids (RLs) are considered as the most popular and efficient biosurfactant, which exhibit superior thermal stability, chemical resistance, low ecotoxicity, biodegradability and good antimicrobial activity. The RLs-producing wild-type strains cannot produce high-yield rhamnolipids to meet with the demand of industrial applications. In view of the problems existing in the practical application of RLs, atmospheric and room temperature plasma-mediated (ARTP) mutagenesis, ultraviolet mutagenesis, ethyl methane sulfonate (EMS) were used alone and combined to mutate and screen Pseudomonas aeruginosa SH6 for many rounds, and a mutant strain P. aeruginosa EZD3 that produced 5.05 g/L of RLs was obtained. Then the original promoter of the rhamnosyl transferase genes was replaced with a strong promoter in strain EZD3, a recombinant strain P. aeruginosa APA1 produced 7.32 g/L of RLs, which showed a 4.67 times increase in RLs production than the original strain P. aeruginosa SH6. Finally, after the optimization of the carbon source of for RLs production, the RLs yield of the recombinant strain P. aeruginosa APA1 reached 7.59 g/L.

摘要

鼠李糖脂(RLs)被认为是最受欢迎且高效的生物表面活性剂,具有卓越的热稳定性、耐化学性、低生态毒性、生物可降解性以及良好的抗菌活性。产RLs的野生型菌株无法高产鼠李糖脂以满足工业应用需求。鉴于RLs实际应用中存在的问题,单独及组合使用常压室温等离子体介导(ARTP)诱变、紫外线诱变、甲基磺酸乙酯(EMS)对铜绿假单胞菌SH6进行多轮诱变和筛选,获得了一株产5.05 g/L RLs的突变株铜绿假单胞菌EZD3。然后在菌株EZD3中用强启动子替换鼠李糖基转移酶基因的原始启动子,构建出一株重组菌株铜绿假单胞菌APA1,其RLs产量达到7.32 g/L,比原始菌株铜绿假单胞菌SH6提高了4.67倍。最后,对重组菌株铜绿假单胞菌APA1的产RLs碳源进行优化后,其RLs产量达到7.59 g/L。

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