福建农林大学学报（自然科学版）2011，Vol.40Issue(2)：165-171,7.
侧孢短芽孢杆菌碱性蛋白酶基因BLG4在枯草芽孢杆菌WB600中的高效表达
Over-expression of an alkaline protease gene BLG4 from Brevibacillus laterosporus G4 in Bacillus subtilis WB600
摘要
Abstract
Protease BLG4 from Brevibacilluslaterosporus G4 has been propesed as an important virulence factor in bacterial pathogenicity against nematodes. In this paper, alkaline protease gene BLG4 from B.laterosporus G4 was cloned and inserted into an expression vector pWT22 to construct a recombinanted plasmid pWT22-BLG4. The pWT22-BLG4 was transformed into Bacillus subtilis WB600 by chemical method. After IPTG ( 1.0 mmol· L-1 ) induction, the protease activity of the fermentation supematant of the transformant pWT22-BLG 4/WB600 reached 620 U · mL-1, which was higher than native BLG4 enzyme activity produced by the wild strain G4. SDS-PAGE analysis revealed that the transformant with recombinanted expression plasmid pWT22-BLG4 had an additional 30 ku protein band, which was identical to B.laterosporus G4. However, the additional band was not present in the negative control Bacillus subtilis WB600. There were no significant differences between the recombinanted and native BLG4 with regard to onzymatic properties. In further bioassays, the recombinanted protease BLG4 had significant nematotoxic activity and ability to degrade nematode cuticles, which indicated that it had a potential as a bio-control agent of plant-parasitic nematodes in the future.关键词
侧孢短芽孢杆菌/侵染性蛋白酶/枯草芽孢杆菌/高效表达/酶学特性/线虫Key words
Brevibacillus laterosporus/ infectious protease/ Bacillus subtilis/ over-expression/ enzymatic property/ nematode分类
生物科学引用本文复制引用
郭菁,田宝玉,蔡婉玲,黄薇,江贤章,黄建忠..侧孢短芽孢杆菌碱性蛋白酶基因BLG4在枯草芽孢杆菌WB600中的高效表达[J].福建农林大学学报（自然科学版）,2011,40(2):165-171,7.基金项目
国家自然科学基金项目(30800735)、福建省自然科学基金(2009J05057、2008F3036)资助. （30800735）
