南极发草一种具有脂肪酶活性的多肽在毕赤酵母载体中的表达

Expression of a Deschampsia antarctica Desv. polypeptide with lipase activity in a Pichia pastoris vector.

作者信息

Rabert Claudia, Gutiérrez-Moraga Ana, Navarrete Alejandro, Navarrete-Campos Darío, Bravo León, Gidekel Manuel

机构信息

Laboratorio de Fisiología y Biología Molecular Vegetal, Instituto de Agroindustria, Facultad de Ciencias Agropecuarias y Forestales, Universidad de La Frontera, Casilla 54D, Temuco 4811230, Chile.

Laboratorio de Fisiología Vegetal, Departamento de Botánica, Universidad de Concepción, Casilla 160-C, Correo 3, Concepción 3801061, Chile.

出版信息

Int J Mol Sci. 2014 Feb 7;15(2):2359-67. doi: 10.3390/ijms15022359.

DOI:10.3390/ijms15022359

PMID:24514564

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3958855/

Abstract

The current study isolated and characterized the Lip3F9 polypeptide sequence of Deschampsia antarctica Desv. (GeneBank Accession Number JX846628), which was found to be comprised of 291 base pairs and was, moreover, expressed in Pichia pastoris X-33 cells. The enzyme was secreted after 24 h of P. pastoris culture incubation and through induction with methanol. The expressed protein showed maximum lipase activity (35 U/L) with an optimal temperature of 37 °C. The lipase-expressed enzyme lost 50% of its specific activity at 25 °C, a behavior characteristic of a psychrotolerant enzyme. Recombinant enzyme activity was measured in the presence of ionic and non-ionic detergents, and a decrease in enzyme activity was detected for all concentrations of ionic and non-ionic detergents assessed.

摘要

本研究分离并鉴定了南极发草（Deschampsia antarctica Desv.）的Lip3F9多肽序列（基因库登录号JX846628），发现其由291个碱基对组成，并且在毕赤酵母X-33细胞中表达。该酶在毕赤酵母培养孵育24小时后通过甲醇诱导分泌。表达的蛋白质在37°C的最佳温度下显示出最大脂肪酶活性（35 U/L）。脂肪酶表达的酶在25°C时失去了50%的比活性，这是耐冷酶的行为特征。在离子和非离子洗涤剂存在的情况下测量重组酶活性，并且在所评估的所有离子和非离子洗涤剂浓度下均检测到酶活性下降。