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嗜酸乳杆菌α-淀粉酶在柠檬明串珠菌中的异源表达与分泌

Heterologous expression and secretion of Lactobacillus amylovorus alpha-amylase in Leuconostoc citreum.

作者信息

Eom Hyun-Ju, Moon Jin-Seok, Seo Eun-Young, Han Nam Soo

机构信息

Department of Food Science and Technology, Research Center for Bioresource and Health, Chungbuk National University, Cheongju, 361-763, South Korea.

出版信息

Biotechnol Lett. 2009 Nov;31(11):1783-8. doi: 10.1007/s10529-009-0079-1. Epub 2009 Jul 18.

Abstract

To develop a gene expression system for Leuconostoc genus, construction of expression vector and expression of a heterologus protein in Leuconostoc was performed. Alpha-amylase gene from Lactobacillus amylovorus was cloned into a Leuconostoc cloning vector, pLeuCM, with its own signal peptide. pLeuCMamy was introduced into Leuconostoc citreum CB2567 and a successful expression of alpha-amy gene was confirmed by enzyme activity assays. About 90% of alpha-amylase activity was detected in the culture broth, revealing most of expressed alpha-amylase was secreted out cells. The signal sequence of alpha-amy gene is a good candidate for the secretion of heterologous protein by using Leuconostoc host-vector system.

摘要

为开发一种用于明串珠菌属的基因表达系统,进行了表达载体的构建以及在明串珠菌中异源蛋白的表达。将来自嗜酸乳杆菌的α-淀粉酶基因克隆到具有自身信号肽的明串珠菌克隆载体pLeuCM中。将pLeuCMamy导入柠檬明串珠菌CB2567,并通过酶活性测定证实了α-淀粉酶基因的成功表达。在培养液中检测到约90%的α-淀粉酶活性,表明大部分表达的α-淀粉酶分泌到细胞外。α-淀粉酶基因的信号序列是利用明串珠菌宿主-载体系统分泌异源蛋白的良好候选序列。

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