Identification of the gene encoding transglutaminase zymogen from Streptomyces hygroscopicus and its expression in Escherichia coli

摘要: OBJECTIVE We identified a microbial transglutaminase (MTGase) gene from Streptomyces hygroscopicus; cloned and expressed it in Escherichia coli. also analyzed the active sites sequence of S. hygroscopicus MTGase through homologous comparison. METHODS Wild-type zymogen (pro-MTGase) was purified liquid culture (CCTCC M203062). N-terminal amino acid this pro-MTGase determined. According to corresponding nucleotide other three species, PCR primers were designed completed amplified sequenced. The sub-cloned into pET-20b(+) vector downstream pelB signal peptide construct expression pET/pro-MTG. RESULTS showed 92% homologue with that platensis caniferus. Rosetta (DE3) pLysS carrying induced IPTG at 24 as extracellular soluble protein. SDS-PAGE recombinant about 44 kDa, similar wild-type hgroscopicus. Recombinant activated trypsin enzyme activity reached 0.24U/mL. CONCLUSION This is first report encoding pro-transglutaminase hygroscopicus, E. coli our country.