毕赤酵母
脂肪酶
生物化学
假丝酵母病
酰基转移酶
乙醇氧化酶
皱纹假丝酵母
毕赤酵母
酵母
异源表达
生物
化学
酶
重组DNA
分子生物学
基因
微生物学
白色念珠菌
作者
Laetitia Brunel,Virginie Neugnot,Laure Landucci,Hélène Boze,Gabrielle Moulin,Frédéric Bigey,Éric Dubreucq
标识
DOI:10.1016/j.jbiotec.2004.03.007
摘要
Candida parapsilosis has been previously shown to produce a lipase/acyltransferase (EC 3.1.1.3) that preferentially catalyses transfer reactions such as alcoholysis over hydrolysis in the presence of suitable nucleophiles other than water, even in aqueous media (aw > 0.9 ). This enzyme has been shown to belong to a new family of lipases. The present work describes the cloning of the gene coding for this lipase/acyltransferase in the yeast Pichia pastoris and the heterologous high-level expression of the recombinant enzyme. The lipase/acyltransferase gene, in which the sequence encoding the signal peptide was replaced by that of the alpha-factor of Saccharomyces cerevisiae, was placed under the control of the methanol inducible promoter of the alcohol oxidase 1 gene (AOX1). A transformed P. pastoris clone, containing five copies of the lipase/acyltransferase gene, was selected for the production of recombinant enzyme. The fed-batch culture supernatant contained 5.8 gl(-1) (weighted) of almost pure recombinant lipase/acyltransferase displaying the same catalytic behavior as the original enzyme.
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