南极假丝酵母脂肪酶B的酿酒酵母表面展示与其催化己酸乙酯的合成.pdfVIP

生 物 工 程 学 报 Chin J Biotech 2008, April 25; 24(4): 673-678 journals.im.ac.cn Chinese Journal of Biotechnology ISSN 1000-3061 cjb@im.ac.cn © 2008 Institute of Microbiology, CAS CSM, All rights reserved 研究简报 B 潘志友, 韩双艳, 林影, 郑穗平 , 510006 : 从南极假丝酵母(Candida antarctica)基因组克隆得到南极假丝酵母脂肪酶B(Candida antarctica Lipase B, CALB) 全基因片段, 利用连接肽celA Linker 将CALB 与酿酒酵母细胞表面展示蛋白α-凝集素的C 端连接融合, 构建表面展示 载体pICAS-celAL-CALB, 转化酵母后获得重组酵母菌Saccharomyces cerevisiae pICAS-celAL-CALB 。该重组酵母菌经 葡萄糖诱导表达及分析, 表明CALB 已在酿酒酵母细胞表面成功展示, 水解活力达26.26 u/(g·dry cell) 。重组酵母菌经冻 干能有效地实现在非水相中全细胞催化己酸和乙醇酯化合成己酸乙酯。反应物己酸与乙醇的摩尔比为 1:1.25, 己酸乙酯 的产率为98.0%, 具有较好的操作稳定性。 : 酵母表面展示, 南极假丝酵母脂肪酶B, 全细胞催化, 己酸乙酯 Expression of Candida antarctica Lipase B on Yeast Surface and Synthesis of Ethyl Hexanoate Catalyzed by CALB Zhiyou Pan, Shuangyan Han, Ying Lin, and Suiping Zheng Key Lab of Fermentation and Enzyme Engineering, School of Bioscience and Technology SCUT, Guangzhou 510006, China Abstract: Short-chain esters play a significant role in the food industry as flavor and aroma constituents. Candida antarctica lipase B (CALB) is one of the most effective catalysts for organic synthesis. We constructed a CALB-displaying yeast whole-cell biocatalyst and applied it to esterification from caproic acid and ethanol. CALB was fused with the α-agglutinin C-terminal and the signal peptide of Glucoamylase in pICAS, a yeast surface display vector, to construct plasmid pICAS-CALB. An extremely Asn-rich linker, named celAL was inserted in t