厦门大学学报（自然科学版）2011，Vol.50Issue(4)：765-771,7.
南极假丝酵母脂肪酶B在毕赤酵母的表达及学性质研究
Expression of Candida antarctica lipase B in Pichia pastoris and Studying of Enzyme Properties
摘要
Abstract
To improve expression efficiency of recombinant Candida antarctica lipase B (CALB) in methylotrophic yeast Pichia pastoris GS115,the DNA sequence encoding CALB was designed and synthesized based on the codon bias of P. Pastoris. The new CALB gene was cloned into pPIC9K,yielding pPIC9K-CALB,and transformed into P. Pastoris GS115. In shake-flask cultivation, the enzyme activity was 46 U/mL after induction by methanol for 120 h. The CALB was purified by metal chelate chromatography by 5. 23 folds. Deglycosylation showed the recombinant CALB was 5 ku bigger than wild type CALB. The optimal pH and temperature of this recombinant enzyme were pH 6. 5 and 30°C, respectively. CALB was relatively stable at temperatures below 40 ℃ within a pH range from 5. 0 to 8. 0. Ca2+ ,Zn2+ ,Mnz+ conduced to enzyme activity, while Cu2+ , Ag+ ,Fe3+ inhibited it.关键词
南极假丝酵母脂肪酶B/毕赤酵母/分泌表达/酶学性质Key words
Candida antarctica lipase B/Pichia pastoris/ secretory expression/enzyme properties分类
生物科学引用本文复制引用
孙金鹏,钱圣一,敬科举,卢英华..南极假丝酵母脂肪酶B在毕赤酵母的表达及学性质研究[J].厦门大学学报（自然科学版）,2011,50(4):765-771,7.基金项目
广东省教育部产学研结合项目(2008B090500218) （2008B090500218）
