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OALib Journal期刊
ISSN: 2333-9721
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β-葡萄糖苷酶基因和内切葡聚糖酶基因在枯草芽孢杆菌中的表达
DOI: 10.3724/SP.J.1145.2013.00990, PP. 990-996
Keywords: 枯草芽孢杆菌,β-葡萄糖苷酶,内切葡聚糖酶,胞外分泌,共表达
Abstract:
为了实现多种纤维素酶基因的分泌型共表达，利用含有枯草芽孢杆菌中性蛋白酶nprb信号肽的穿梭质粒pp43jm2作为表达载体，将来源于热纤梭菌的内切葡聚糖酶基因cela与来源于多粘芽孢杆菌的β-葡萄糖苷酶基因bgla和bglb在枯草芽孢杆菌wb800中进行了分泌型的单独表达及共表达.结果表明，内切葡聚糖酶基因cela与β-葡萄糖苷酶基因bgla和bglb均能够在枯草芽孢杆菌中实现分泌表达，而且检测到了cela和bglb基因在枯草芽孢杆菌中的分泌型共表达，其中，共表达菌株的胞外酶液分别与pasc和纤维二糖底物反应时，释放出674mg/l和24mg/l的葡萄糖.而bgla基因只检测到在胞内表达，其胞内酶液以纤维二糖为底物时生成938.7mg/l的葡萄糖.本研究为实现在枯草芽孢杆菌中纤维素酶多组分人工组装并为集成生物工艺菌种的构建奠定了一定的实验基础.
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