title: Construction and development of a novel expression system of Streptomyces.
abstract: 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.
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu 214122, China.
authors.forename: Chengran
authors.initials: C
authors.lastname: Guan
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu 214122, China.
authors.forename: Wenjing
authors.initials: W
authors.lastname: Cui
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu 214122, China.
authors.forename: Xiaotian
authors.initials: X
authors.lastname: He
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu 214122, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Hu
authors.affiliation: School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200030, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Xu
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu 214122, China.
authors.forename: Guocheng
authors.initials: G
authors.lastname: Du
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu 214122, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.affiliation: School of Biotechnology, Key Laboratory of Industrial Biotechnology (Ministry of Education), Jiangnan University, Wuxi, Jiangsu 214122, China. Electronic address: zhmzhou@jiangnan.edu.cn.
authors.forename: Zhemin
authors.initials: Z
authors.lastname: Zhou
mesh_terms: D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012248:Rhodotorula / Q000201:enzymology / Q000235:genetics; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins
keywords: Expression system; Heterologous protein expression; Streptomyces; TGase