食品科学2017，Vol.38Issue(8)：69-73,5.DOI:10.7506/spkx1002-6630-201708012
高产β-葡萄糖苷酶工程菌株的构建及其在2,6-二甲氧基对苯醌发酵制备中的应用
Construction of an Engineered Strain Producing High β-Glucosidase Activity and Its Application in the Production of 2,6-Dimethoxybenzoquinone
摘要
Abstract
An engineered strain producing high β-glucosidase activity was constructed and applied in the production of 2,6-dimethoxybenzoquinone.The β-glucosidase gene was amplified from the genomic DNA of Saccharomyces cerevisiae CS1401 using PCR.The amplified gene was then ligated to pPICZαA vector and transformed to Pichia pastoris X33 for expression under the induction of methanol.The activity of the recombinant enzyme was measured,and S.cerevisiae CS1401 and the recombinant strain were separately cultured in wheat germ medium for the production of 2,6-dimethoxybenzoquinone.The results indicated that the β-glucosidase gene of S.cerevisiae CS1401 was efficiently expressed in the engineered strain ofP.pastoris X33,giving a β-glucosidase activity of up to 4.5 U/mL,which was 5 times higher than that of S.cerevisiae CS1401.Besides,the yield of 2,6-dimethoxybenzoquinone produced by the recombinant strain in shaking flasks reached 935.7 tg/g,which was 40％ higher than that of S.cerevisiae CS1401.Therefore,this engineered strain has a promising prospect of application in the microbial production of 2,6-dimethoxybenzoquinone.关键词
酿酒酵母/β-葡萄糖苷酶/毕赤酵母/高效表达/2,6-二甲氧基对苯醌Key words
Saccharomyces cerevisiae/β-glucosidase/Pichia pastoris/over-expression/2,6-dimethoxybenzoquinone分类
生物科学引用本文复制引用
易晓男,任清华,程炜,孙姜,齐斌,王立梅..高产β-葡萄糖苷酶工程菌株的构建及其在2,6-二甲氧基对苯醌发酵制备中的应用[J].食品科学,2017,38(8):69-73,5.基金项目
江苏省重点研发计划项目(BE2014325) （BE2014325）
