食品科学技术学报2019，Vol.37Issue(2)：36-41,6.DOI:10.3969/j.issn.2095-6002.2019.02.006
烟曲霉脂肪酶B在毕赤酵母中的表达及酶学性质研究
Expression of Aspergillus fumigates Lipase B in Pichia pastoris and Biochemical Characterization
摘要
Abstract
A lipase gene (AFLB) was cloned from Aspergillus fumigatus GIM3. 20. Sequence alignment analysis showed that the AFLB shared 31% amino acid sequences identities with that of Candida antarctica lipase B ( CALB) , and both of them were belonged to the CALB-like family. AFLB contained a similar conserved pentapeptide ( SWSQG) that the member of CALB-like family shared. Its catalytic triad was supposed to be made up by residues S233,D318, and H361, and the catalytic serine locates in the pen-tapeptide. What’s more, AFLB hosted a special N-terminal extended sequence, which was a feature of Aspergillus spp. lipase B. AFLB was successfully expressed in Pichia pastoris and purified by affinity chromatography. The result of SDS-PAGE showed that AFLB had partial N-glycosylation, displaying molecular weights of 45-66 kDa and 42 kDa. Characterization of AFLB lipase showed that the optimum reaction pH was 7. 0 and the optimum reaction temperature was 40℃. The optimum substrate for AFLB was p-nitrophenol myristate. AFLB was the homologue of lipase CALB, and might have good industrial application prospect.关键词
烟曲霉/脂肪酶/重组表达/毕赤酵母/酶学性质Key words
Aspergillus fumigates/lipase/recombinant expression/Pichia pastoris/enzymatic property分类
轻工纺织引用本文复制引用
唐薇,黄伟谦,杨博,王永华,蓝东明..烟曲霉脂肪酶B在毕赤酵母中的表达及酶学性质研究[J].食品科学技术学报,2019,37(2):36-41,6.基金项目
广东省省级科技计划项目(017A050503001). （017A050503001）
