中国食品添加剂Issue(2)：81-86,6.
多黏芽孢杆菌产α-环糊精葡萄糖基转移酶在大肠杆菌中的可溶性表达及其转化产物特异性研究
Study of soluble expression in Escherichia coli of α-cyclodextrin glycosyl transferase produced by Paenibacillus macerans and properties of its derivatives
摘要
Abstract
The gene encoding α-cyclodextrin glycosyltransferase (α-CGTase) was amplified by degenerate primer from Paenibacillus macerans YLW and was inserted into expression vector pET28a (+),and a recombinant strain α-CGTase-pET28a/BL21 (DE3) was constructed.After induction for 15h at 16℃ with 1 mM IPTG,α-CGTase was expressed in solution and the activity of α-CGTase in the periplasm was reached to 10046 U/mL,which was approximately 3.25-fold with that from the parent strain.The recombinant α-CGTase was purified by one-step nickel affinity chromatography,and purification fold and yield was 6.05 and 28.82％,respectively.The enzyme catalytic tests demonstrated that the yield of total CD reached 40.7％ after 15h incubation with 5％ potato starch,and α-CD:β-CD:γ-CD ratio was 43.6:41.8:14.6.Therefore,the recombinant α-CGTase showed better special enzymatic characterization and have potential utilizaton in industrial production of α-CD with further op timized transformation condition in the future.关键词
α-环糊精葡萄糖基转移酶/可溶性表达/纯化/转化性质Key words
α-cyclodextrin glycosyltransferase/soluble expression/purification/characterization of transformation分类
生物科学引用本文复制引用
王琰,李皎,杨国武,王军,邓媛,万一..多黏芽孢杆菌产α-环糊精葡萄糖基转移酶在大肠杆菌中的可溶性表达及其转化产物特异性研究[J].中国食品添加剂,2017,(2):81-86,6.基金项目
陕西省科学院应用基础研究项目(2013K-09 （）
陕西省科学院科学技术平台项目(2015k-33). （2015k-33）
