Guan Chengran, Cui Wenjing, He Xiaotian, Hu Xu, Xu Jun, Du Guocheng, Chen Jian, Zhou Zhemin
School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu 214122, China.
School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200030, China.
Protein Expr Purif. 2015 Sep;113:17-22. doi: 10.1016/j.pep.2015.04.009. Epub 2015 May 6.
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 (SP(tg)) of transglutaminase (TGase) from Streptomyces hygroscopicus WSH03-13 into vector pIJ86, 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 SP(tg), 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.
众所周知，链霉菌是一种极具吸引力的宿主，可将大量具有强大生物活性的蛋白质分泌到培养上清液中。为了扩展其表达系统，我们构建了一种新型表达质粒用于链霉菌中的基因表达，方法是将来自吸水链霉菌WSH03 - 13的转谷氨酰胺酶（TGase）的启动子（P(tg)）和信号肽（SP(tg)）插入载体pIJ86，随后连接多克隆位点和转录终止子fd（fd-ter）。通过优化信号肽酶切割位点和SP(tg)的稀有密码子，进一步增强了该载体的分泌能力，得到表达载体pSG02。使用该载体，TGase在几种链霉菌菌株的上清液中得到了高效且大量的表达。此外，枯草芽孢杆菌Zj016的氨肽酶（BSAP）和粘红酵母的苯丙氨酸解氨酶（PAL）等异源蛋白也通过该载体在多种链霉菌菌株中得到了表达。该表达系统对于其他蛋白质的表达应该是有用的。
