文献类型：期刊文献

英文题名：Construction of a Shuttle Vector for Protein Secretory Expression in Bacillus subtilis and the Application of the Mannanase Functional Heterologous Expression

作者：Guo, Su[1];Tang, Jia-jie[1];Wei, Dong-zhi[1];Wei, Wei[1]

机构：[1]E China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2014

卷号：24

期号：4

起止页码：431

外文期刊名：JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000335092400001)】；

基金：This research was financially supported by the Fundamental Research Funds for the Central Universities of China (No. WF1214047), the National High Technology Research and Development Program of China (No. 2013AA102109), and National Major Science and Technology Projects of China (No. 2012ZX09304009).

语种：英文

外文关键词：Bacillus subtilis; Bacillus pumilus; green fluorescent protein; mannanase; P43 promoter; secretory expression

摘要：We report the construction of two Bacillus subtilis expression vectors, pBNS1/pBNS2. Both vectors are based on the strong promoter P43 and the ampicillin resistance gene expression cassette. Additionally, a fragment with the Shine-Dalgarno sequence and a multiple cloning site (BamHI, SalI, SacI, XhoI, PstI, SphI) were inserted. The coding region for the amyQ (encoding an amylase) signal peptide was fused to the promoter P43 of pBNS1 to construct the secreted expression vector pBNS2. The applicability of vectors was tested by first generating the expression vectors pBNS1-GFP/pBNS2-GFP and then detecting for green fluorescent protein gene expression. Next, the mannanase gene from B. pumilus Nsic-2 was fused to vector pBNS2 and we measured the mannanase activity in the supernatant. The mannanase total enzyme activity was 8.65 U/ml, which was 6 times higher than that of the parent strain. Our work provides a feasible way to achieve an effective transformation system for gene expression in B. subtilis and is the first report to achieve B. pumilus mannanase secretory expression in B. subtilis.