Engineered global regulator H-NS improves the acid tolerance of E. coli

被引:32
作者
Gao, Xianxing [1 ]
Yang, Xiaofeng [2 ]
Li, Jiahui [2 ]
Zhang, Yan [1 ,3 ]
Chen, Ping [2 ]
Lin, Zhanglin [1 ,2 ]
机构
[1] Tsinghua Univ, Dept Chem Engn, One Tsinghua Garden Rd, Beijing 100084, Peoples R China
[2] South China Univ Technol, Sch Biol & Biol Engn, 382 East Outer Loop Rd,Univ Pk, Guangzhou 510006, Guangdong, Peoples R China
[3] China Acad Agr Sci, Shenzhen Agr Genom Inst, 7 Pengfei Rd, Shenzhen 518120, Guangdong, Peoples R China
关键词
Acid tolerance; E; coli; H-NS; Global transcription machinery engineering (gTME); Error-prone PCR; NUCLEOID-ASSOCIATED PROTEIN; ESCHERICHIA-COLI; TRANSCRIPTION MACHINERY; STRESS-RESPONSE; DOMAIN ORGANIZATION; ETHANOL TOLERANCE; LACTIC-ACID; RESISTANCE; GROWTH; RPOS;
D O I
10.1186/s12934-018-0966-z
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
Background: Acid stress is often encountered during industrial fermentation as a result of the accumulation of acidic metabolites. Acid stress increases the intracellular acidity and can cause DNA damage and denaturation of essential enzymes, thus leading to a decrease of growth and fermentation yields. Although acid stress can be relieved by addition of a base to the medium, fermentations with acid-tolerant strains are generally considered much more efficient and cost-effective. Results: In this study, the global regulator H-NS was found to have significant influence on the acid tolerance of E. coil. The final OD600 of strains overexpressing H-NS increased by 24% compared to control, when cultured for 24 h at pH 4.5 using HCI as an acid agent. To further improve the acid tolerance, a library of H-NS was constructed by error-prone PCR and subjected to selection. Five mutants that conferred a significant growth advantage compared to the control strain were obtained. The final OD600 of strains harboring the five H-NS mutants was enhanced by 26-53%, and their survival rate was increased by 10- to 100-fold at pH 2.5. Further investigation showed that the improved acid tolerance of H-NS mutants coincides with the activation of multiple acid resistance mechanisms, in particular the glutamate- and glutamine-dependent acid resistance system (AR2). The improved acid tolerance of H-NS mutants was also demonstrated in media acidified by acetic acid and succinic acid, which are common acidic fermentation by-products or products. Conclusions: The results obtained in this work demonstrate that the engineering of H-NS can enhance the acid tolerance of E. coli. More in general, this study shows the potential of the engineering of global regulators acting as repressors, such as H-NS, as a promising method to obtain phenotypes of interest. This approach could expand the spectrum of application of global transcription machinery engineering.
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