工业微生物2017，Vol.47Issue(1)：17-23,7.DOI:10.3969/j.issn.1001-6678.2017.01.003
通过信号肽筛选优化耐高温α-淀粉酶在枯草芽孢杆菌中的分泌
Optimization of thermostable α-amylase secretion by screening of optimal signal peptide in Bacillus subtilis
摘要
Abstract
A screening vector for signal peptides from B.subtilis was constructed using a POE-PCR cloning method.The thermostable a-amylase from Geobacillus stearothermophilus NUB3621 was chosen as the target gene and cloned into E.coli and B.subtilis shuttle vector pMA5.46 signal peptides was amplified from the genome of B.subtilis 168.The DNA multimer was generated based on each signal peptide and the linear vector backbone by prolonged overlap extension PCR was directly transformed into B.subtilis 1A751,then 46 strains,each amylase protein was driven by different signal peptide,were cultured.The fermentation results revealed that except 5 signal peptides showed inappropriate fit with desired secretion target protein,the others showed different ability of the export of α-amylase in B.subtilis.The highest enzyme activity of 1 393.3 U/mL was detected from the recombinant strain containing bgls signal peptide.关键词
耐高温/淀粉酶/信号肽/枯草芽孢杆菌Key words
thermostable/amylase/signal peptide/B.subtilis引用本文复制引用
刘金岚,付刚,董会娜,袁向华,张大伟..通过信号肽筛选优化耐高温α-淀粉酶在枯草芽孢杆菌中的分泌[J].工业微生物,2017,47(1):17-23,7.基金项目
本课题受国家自然科学基金(21506244,31370089)项目及天津市科技支撑计划重点项目(14ZCZDSY00065)、天津市自然科学基金(16JCYBJC23500,15JCQNJC09500)支持. （21506244,31370089）
