Improvement of lichenysin production in Bacillus licheniformis by replacement of native promoter of lichenysin biosynthesis operon and medium optimization

被引:71
|
作者
Qiu, Yimin [1 ]
Xiao, Fang [1 ]
Wei, Xuetuan [3 ]
Wen, Zhiyou [3 ,4 ]
Chen, Shouwen [1 ,2 ]
机构
[1] Huazhong Agr Univ, Coll Life Sci & Technol, State Key Lab Agr Microbiol, Wuhan 430070, Peoples R China
[2] Hubei Univ, Coll Life Sci, Hubei Collaborat Innovat Ctr Green Transformat Bi, Wuhan 430062, Peoples R China
[3] Huazhong Agr Univ, Coll Food Sci & Technol, Wuhan 430070, Peoples R China
[4] Iowa State Univ, Dept Food Sci & Human Nutr, Ames, IA 50011 USA
基金
中国国家自然科学基金;
关键词
Lichenysin; Bacillus licheniformis; Promoter replacement; Medium optimization; MEDIUM COMPONENTS; SURFACE-ACTIVITY; HIGH-LEVEL; SUBTILIS; BIOSURFACTANT; LIPOPEPTIDES; EXPRESSION; OVEREXPRESSION; MYCOSUBTILIN; MEGATERIUM;
D O I
10.1007/s00253-014-5978-y
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Lichenysin is a biodegradable surfactant with huge potential for recovering crude oil from the oil reservoir. The current production of lichenysin is made through fermentation from wild strain of Bacillus licheniformis, which is limited by low yield. The aim of this work was to improve lichenysin-producing capability of a wide strain B. licheniformis WX-02. Lichenysin produced from WX-02 was first extracted, purified, and identified. Through the substitution of the promoter of lichenysin biosynthesis operon, the mutants B. licheniformis WX02-P43lch, WX02-Pxyllch, and WX02-Psrflch were constructed with the constitutive promoter (P43), the xylose-inducible promoter (P (xyl) ), and the surfactin operon promoter (P (srf) ), respectively. A consistent change trend was observed between lichenysin production and lchAA gene transcription, confirming the strength of the promoters as an important factor for lichenysin synthesis. Among the three mutants, WX02-Psrflch produced the highest lichenysin yield. The production by the mutant WX02-Psrflch was further improved with the optimization of the major medium components including glucose, NH4NO3, and Na2HPO4/KH2PO4. Under 30 g/L glucose, 5 g/L NH4NO3, and 80 mM/60 mM Na2HPO4/KH2PO4, the strain WX02-Psrflch produced 2,149 mg/L lichenysin, a 16.8-fold improvement compared to that of wild strain WX-02.
引用
收藏
页码:8895 / 8903
页数:9
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