作　　者： ; ; ; ; ;

机构地区： 华南理工大学生物科学与工程学院

出　　处： 《现代食品科技》 2013年第7期1477-1481,共5页

摘　　要： 本研究将来自抗辐射不动杆菌CMC-1的碱性脂肪酶基因去掉信号肽,经密码子优化及全基因合成后克隆到pPICZαA载体,构建了重组质粒pPICZαA-ARL,质粒线性化后转化毕赤酵母X33,筛选得到分泌表达碱性脂肪酶的重组毕赤酵母X33/pPICZαA-ARL。摇瓶发酵液上清酶活最高达65 U/mL,初步研究了该脂肪酶的酶学性质,其最适作用温度为50℃,最适pH为9.0,最适底物是对硝基苯酚辛酸酯。 Lipase is one of the most important industrial enzymes applied widely in food,textile,papermaking and pharmaceutical industries.The alkaline lipase gene from Acinetobacter radioresistens CMC-1 was synthesized with its own signal peptide removed followed by codon optimization,and then cloned into the secreted expression vector pPICZαA.The constructed plasmid was named as pPICZαA-ARL.The recombinant plasmid was used to transform into the Pichia pastoris X33.One of the best transformants harboring ARL integrated into the P.pastoris genomic DNA was cultured in shake flask.The maximum activity of the culture supernatant was 65 U/mL.The properties of the recombinant lipase was partial characterized.The results showed that the recombinant lipase had an optimal activity at 50 ℃ and pH 9.0 and the optimum substrate of recombinant ARL was 4-nitrophenyl caprylate,which seemed a little difference from the wild one.

关 键 词： 抗辐射不动杆菌 碱性脂肪酶 密码子优化 毕赤酵母 分泌表达

领　　域： [生物学]