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OALib Journal期刊
ISSN: 2333-9721
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β-甘露聚糖酶基因在枯草芽孢杆菌中的克隆及表达
DOI: 10.3724/SP.J.1145.2012.00672, PP. 672-677
Keywords: β-甘露聚糖酶,枯草芽孢杆菌,信号肽,克隆,表达,重组菌株,酶学性质
Abstract:
从bacillussubtilisjna3-10中克隆出β-甘露聚糖酶基因成熟肽链编码序列mana1和含信号肽的β-甘露聚糖酶基因mana2，在b.subtilis168中克隆表达，分别筛选获得高效分泌表达β-甘露聚糖酶的重组菌株bpm1001（pma5-mana1/b.subtilis168）和bpm1002（pma5-mana2/b.subtilis168），结果表明菌株bpm1002总酶活力是菌株bpm1001的9.65倍，是原始菌株的13.1倍.在基因mana2下游引入his序列克隆出β-甘露聚糖酶基因mana3，获得枯草芽孢杆菌168重组菌株bpm1003.采用ni-nta柱纯化重组菌株bpm1003分泌表达的β-甘露聚糖酶，并研究其酶学性质，该酶促反应的最适ph为6.5，最适温度为65℃，在37℃条件下保存一个月酶活力依然保留有77.8%.5l发酵罐放大实验结果表明魔芋粉对于产β-甘露聚糖酶具有明显的诱导作用，酶活力最高可达2748.82u/ml.图9表3参19
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