title: Enhancement of a high efficient autoinducible expression system in Bacillus subtilis by promoter engineering.
abstract: Quorum-sensing related promoter srfA (PsrfA) was used to construct autoinducible expression system for production of recombinant proteins in Bacillus subtilis. PsrfA was prominent in the unique property of inducer-free activity that is closely correlated with cell density. Here, using green fluorescent protein (GFP) as the reporter protein, PsrfA was optimized by shortening its sequences and changing the nucleotides at the conserved regions of -35 -15 and -10 regions, obtaining a library of PsrfA derivatives varied in the strength of GFP production. Among all the promoter mutants, the strongest promoter P10 was selected and the strength in GFP expression was 150% higher than that of PsrfA. Heterologous protein of aminopeptidase and nattokinase could be overexpressed by P10, the activities of which were 360% and 50% higher than that of PsrfA, respectively. These results suggested that the enhanced promoter P10 could be used to develop autoinducible expression system for overexpression of heterologous proteins in B. subtilis.
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu, 214122, China.
authors.forename: Jintao
authors.initials: J
authors.lastname: Cheng
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu, 214122, China.
authors.forename: Chengran
authors.initials: C
authors.lastname: Guan
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu, 214122, China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Cui
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu, 214122, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhou
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu, 214122, China.
authors.forename: Zhongmei
authors.initials: Z
authors.lastname: Liu
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu, 214122, China. Electronic address: wjlee01@gmail.com.
authors.forename: Weijiang
authors.initials: W
authors.lastname: Li
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu, 214122, China. Electronic address: zhmzhou@jiangnan.edu.cn.
authors.forename: Zhemin
authors.initials: Z
authors.lastname: Zhou
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D009154:Mutation; D010453:Peptide Synthases / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D010453:Peptide Synthases; C069542:surfactin synthetase
keywords: Autoinducible; Expression system; Overexpression; Promoter