α-淀粉酶在不同地衣芽孢杆菌宿主菌中分泌表达的对比分析

doi:10.7506/spkx1002-6630-201517037

食品科学

α-淀粉酶在不同地衣芽孢杆菌宿主菌中分泌表达的对比分析

陈敬帮1，周银华1，赵新宇1,2，陈守文1，魏雪团1,2,*

1.华中农业大学农业微生物学国家重点实验室，湖北武汉 430070；
2.华中农业大学食品科学技术学院，湖北武汉 430070

出版日期:2015-09-15 发布日期:2015-09-11

Comparative Analysis on the Effects of Different Host Strains of Bacillus licheniformis on the Expression and Secretion of α-Amylase

CHEN Jingbang1, ZHOU Yinhua1, ZHAO Xinyu1,2, CHEN Shouwen1, WEI Xuetuan1,2,*

1. State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, China;
2. College of Food Science and Technology, Huazhong Agricultural University, Wuhan 430070, China

Online:2015-09-15 Published:2015-09-11

摘要/Abstract

摘要：

研究不同地衣芽孢杆菌（Bacillus licheniformis）宿主菌对α-淀粉酶分泌表达的影响。以Bacillus subtilis的P43启动子，B. licheniformis WX-02 α-淀粉酶基因的信号肽、编码区和终止子序列为表达原件，构建了α-淀粉酶分泌表达载体pP43SAT。将pP43SAT分别转入3 株B. licheniformis宿主菌：WX-02（ΔamyL）、BL9和BL10，获得3 株α-淀粉酶基因工程菌WX-02（ΔamyL，pP43SAT）、BL9（pP43SAT）和BL10（pP43SAT），并将构建的3 株工程菌进行发酵对比分析。BL9（pP43SAT）和BL10（pP43SAT）的淀粉酶发酵活力分别达到94.01 U/mL和101.94 U/mL，比原始宿主菌WX-02（ΔamyL，pP43SAT）分别提高了21%和31%，这说明BL9和BL10新型宿主菌有利于淀粉酶的分泌表达。本研究证明多蛋白酶基因缺失可显著提高α-淀粉酶的表达，为α-淀粉酶的高效表达提供了新型宿主菌和新策略。

关键词: &alpha, -淀粉酶, 地衣芽孢杆菌, 宿主菌, 分泌表达

Abstract:

In the present study, effects of different host strains of Bacillus licheniformis on the expression and secretion
of α-amylase were investigated. Firstly, the α-amylase expression vector, named pP43SAT, was constructed based on
the promoter P43 from Bacillus subtilis and the signal peptide, coding region and terminator of α-amylase gene from
B. licheniformis WX-02. The pP43SAT plasmids were then transformed into the B. licheniformis host strains WX-02
(ΔamyL), BL9 and BL10B, respectively, to obtain the α-amylase genetic engineering strains WX-02 (ΔamyL; pP43SAT),
BL9 (pP43SAT) and BL10 (pP43SAT), respectively. The α-amylase fermentation processes of these engineered strains
were compared. The α-amylase activities of strains BL9 (pP43SAT) and BL10 (pP43SAT) reached 94.01 and 101.94 U/mL,
respectively, which were increased by 21% and 31%, respectively, as compared with that of WX-02 (ΔamyL; pP43SAT).
These results showed that the novel host strains BL9 and BL10 contributed to the secretion and expression of α-amylase.
It was proved that the deletion of multiple proteases genes could enhance α-amylase expression obviously, and this study
provided the novel host strains and novel strategies for efficient expression of α-amylase.

Key words: α-amylase, Bacillus licheniformis, host strain, secretion and expression

中图分类号:

Q812

陈敬帮，周银华，赵新宇，陈守文，魏雪团. α-淀粉酶在不同地衣芽孢杆菌宿主菌中分泌表达的对比分析[J]. 食品科学, doi: 10.7506/spkx1002-6630-201517037.

CHEN Jingbang, ZHOU Yinhua, ZHAO Xinyu, CHEN Shouwen, WEI Xuetuan. Comparative Analysis on the Effects of Different Host Strains of Bacillus licheniformis on the Expression and Secretion of α-Amylase[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201517037.

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α-淀粉酶在不同地衣芽孢杆菌宿主菌中分泌表达的对比分析

Comparative Analysis on the Effects of Different Host Strains of Bacillus licheniformis on the Expression and Secretion of α-Amylase

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