%0 Journal Article
%T Identification of the gene encoding transglutaminase zymogen from Streptomyces hygroscopicus and its expression in Escherichia coli<br>Streptomyces hygroscopicus谷氨酰胺转胺酶酶原基因的鉴定及在大肠杆菌中的表达
%A Zengliang Ren
%A Dongxu Zhang
%A Meiying Yu
%A Qingxin Zhao
%A Guocheng Du
%A Jian Chen
%A Jing Wu
%A <br>任增亮
%A 张东旭
%A 俞梅英
%A 赵庆新
%A 堵国成
%A 陈坚
%A 吴敬
%J 微生物学报
%D 2008
%I 
%X OBJECTIVE: We identified a microbial transglutaminase (MTGase) gene from Streptomyces hygroscopicus; cloned and expressed it in Escherichia coli. We also analyzed the active sites sequence of S. hygroscopicus MTGase through homologous sequence comparison. METHODS: Wild-type microbial transglutaminase zymogen (pro-MTGase) was purified from liquid culture of S. hygroscopicus (CCTCC M203062). N-terminal amino acid sequence of this pro-MTGase was determined. According to the N-terminal sequence and the corresponding nucleotide sequence of MTGase from other three Streptomyces species, PCR primers of S. hygroscopicus pro-MTGase were designed and the completed gene of pro-MTGase was amplified and sequenced. The gene was sub-cloned into pET-20b(+) vector downstream pelB signal peptide to construct the expression vector pET/pro-MTG. RESULTS: The nucleotide sequence showed 92% homologue with that of S. platensis and S. caniferus. Rosetta (DE3) pLysS carrying the expression vector was induced with IPTG at 24 and expressed pro-MTGase as extracellular soluble protein. SDS-PAGE showed the expressed recombinant pro-MTGase was about 44 kDa, similar to the wild-type pro-MTGase purified from S. hgroscopicus. Recombinant pro-MTGase was activated with trypsin and the enzyme activity reached to 0.24U/mL. CONCLUSION: This is the first report of the gene encoding microbial pro-transglutaminase from S. hygroscopicus, and also this is the first report of expression extracellular soluble pro-MTGase in E. coli in our country.
%K Streptomyces hygroscopicus
%K microbial transglutaminase
%K E
%K coli
%K cloning
%K expression<br>吸水链霉菌
%K 谷氨酰胺转胺酶
%K 大肠杆菌
%K 克隆
%K 表达
%K Streptomyces
%K hygroscopicus
%K 谷氨酰胺
%K 转胺酶
%K 酶原
%K 基因同源性
%K 大肠杆菌
%K 可溶性表达
%K Escherichia
%K coli
%K expression
%K transglutaminase
%K encoding
%K gene
%K 酶活
%K 活化
%K 胰蛋白酶
%K 重组蛋白
%K 原相
%K 分子量
%K 显示
%K 胞外
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=1EDC6F729B2D4EF3B63EA3EDBBDE8500&yid=67289AFF6305E306&vid=B6DA1AC076E37400&iid=E158A972A605785F&sid=03436AC72A659ACA&eid=5CB576B96D187F64&journal_id=0001-6209&journal_name=微生物学报&referenced_num=1&reference_num=19