从皱褶假丝酵母中分离羧酸酯脂肪酶（CEL）同工酶并鉴定相应基因。

Isolation of carboxylester lipase (CEL) isoenzymes from Candida rugosa and identification of the corresponding genes.

作者信息

Diczfalusy M A, Hellman U, Alexson S E

机构信息

Department of Medical Laboratory Sciences and Technology, Karolinska Institutet, Huddinge University Hospital, Sweden.

出版信息

Arch Biochem Biophys. 1997 Dec 1;348(1):1-8. doi: 10.1006/abbi.1997.0382.

DOI:10.1006/abbi.1997.0382

PMID:9390168

Abstract

The yeast Candida rugosa produces extracellular lipases which are widely used for industrial purposes. A commercial lipase preparation from this yeast can be separated into several isoenzymes which differ in carbohydrate content, isolelectric point, substrate specificity, and primary sequence. We have here purified and characterized three lipases, which also hydrolyze p-nitrophenyl esters, from a commercial preparation of this yeast. These three carboxylester lipases (CELs) elute differently on hydrophobic interaction chromatography, and have different carbohydrate contents and substrate specificities. Sequence analysis of their amino termini and peptides generated by LysC treatment showed that CEL-1 and CEL-3 probably have identical primary structure while CEL-2 was proven to be a different enzyme. Sequence comparison showed that both CEL-1 and CEL-3 are products of the LIP1 gene and that CEL-2 is the gene product of LIP2, cloned by Longhi et al. (Biochim. Biophys. Acta 1131, 227-232, 1992).

摘要

皱落假丝酵母能产生广泛应用于工业用途的胞外脂肪酶。从这种酵母中制备的一种商业脂肪酶制剂可分离成几种同工酶，它们在碳水化合物含量、等电点、底物特异性和一级序列上存在差异。我们在此从这种酵母的一种商业制剂中纯化并鉴定了三种也能水解对硝基苯酯的脂肪酶。这三种羧酸酯脂肪酶（CELs）在疏水相互作用色谱上的洗脱方式不同，且碳水化合物含量和底物特异性也不同。对它们的氨基末端以及经LysC处理产生的肽段进行序列分析表明，CEL-1和CEL-3可能具有相同的一级结构，而CEL-2被证明是一种不同的酶。序列比较表明，CEL-1和CEL-3都是LIP1基因的产物，而CEL-2是Longhi等人（《生物化学与生物物理学报》1131, 227 - 232, 1992）克隆的LIP2基因的产物。