Construction and development of a novel expression system of Streptomyces
Construction and development of a novel expression system of Streptomyces
复制标题
DOI：
10.1016/j.pep.2015.04.009
复制
发表时间：
2015-09-01
影响因子：
1.6
通讯作者：
Zhou, Zhemin
中科院分区：
文献类型：
--
作者：
Guan, Chengran;Cui, Wenjing;Zhou, Zhemin
众所周知，链霉菌是一种有吸引力的宿主，可以在培养上清中产生大量具有强生物活性的蛋白质。为了扩大其表达体系，我们将吸湿链霉菌WSH03-13中转谷氨酰胺酶（TGase）启动子（P-tg）和信号肽（SPtg）插入到载体086中，构建了一种新的链霉菌基因表达质粒，随后插入多个克隆位点和转录终止子fd （fd-ter）。通过优化信号肽酶裂解位点和SPtg的一个罕见密码子，进一步增强了载体的分泌能力，得到表达载体pSG02。利用该载体，TGase在数株链霉菌的上清液中得到了高效表达。此外，来自枯草芽孢杆菌Zj016的异源蛋白氨基肽酶（BSAP）和来自粘红酵母（PAL）的苯丙氨酸解氨酶也通过该载体在各种链霉菌中表达。该表达系统可用于其他蛋白的表达。(C) 2015爱思唯尔公司版权所有。
Streptomyces is well known to be an attractive host for producing large amounts of proteins with potent biological activities into the culture supernatant. To expand its expression system, we constructed a novel expression plasmid for gene expression in Streptomyces by inserting the promoter (P-tg) and the signal peptide (SPtg) of transglutaminase (TGase) from Streptomyces hygroscopicus WSH03-13 into vector 086, followed by multiple cloning sites and a transcriptional terminator fd (fd-ter). The secretion capacity of the vector was further enhanced by optimizing the signal peptidase cleavage site and a rare codon of SPtg, yielding expression vector pSG02. Using this vector, TGase was actively and greatly expressed in the supernatant in several Streptomyces strains. In addition, the heterologous proteins aminopeptidase from Bacillus subtilis Zj016 (BSAP) and phenylalanine ammonia-lyase from Rhodotorula glutinis (PAL) were also expressed in various Streptomyces strains by this vector. This expression system should be useful for the expression of other proteins. (C) 2015 Elsevier Inc. All rights reserved.