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中国精品科技期刊2020
张莹莹, 石楠, 杜萍萍, 申培立, 李志辉, 于宏伟, 卢海强, 苏旭东, 张伟, 檀建新. 一株产谷氨酰胺转氨酶菌株的分离、鉴定及其tgl的克隆表达[J]. 食品工业科技, 2015, (11): 141-146. DOI: 10.13386/j.issn1002-0306.2015.11.020
引用本文: 张莹莹, 石楠, 杜萍萍, 申培立, 李志辉, 于宏伟, 卢海强, 苏旭东, 张伟, 檀建新. 一株产谷氨酰胺转氨酶菌株的分离、鉴定及其tgl的克隆表达[J]. 食品工业科技, 2015, (11): 141-146. DOI: 10.13386/j.issn1002-0306.2015.11.020
ZHANG Ying-ying, SHI Nan, DU Ping-ping, SHEN Pei-li, LI Zhi-hui, YU Hong-wei, LU Hai-qiang, SU Xu-dong, ZHANG Wei, TAN Jian-xin. Isolation and identification of a transglutaminase producing strain and its tgl cloning and expression[J]. Science and Technology of Food Industry, 2015, (11): 141-146. DOI: 10.13386/j.issn1002-0306.2015.11.020
Citation: ZHANG Ying-ying, SHI Nan, DU Ping-ping, SHEN Pei-li, LI Zhi-hui, YU Hong-wei, LU Hai-qiang, SU Xu-dong, ZHANG Wei, TAN Jian-xin. Isolation and identification of a transglutaminase producing strain and its tgl cloning and expression[J]. Science and Technology of Food Industry, 2015, (11): 141-146. DOI: 10.13386/j.issn1002-0306.2015.11.020

一株产谷氨酰胺转氨酶菌株的分离、鉴定及其tgl的克隆表达

Isolation and identification of a transglutaminase producing strain and its tgl cloning and expression

  • 摘要: 用稀释平板法从土壤样品中分离到166株细菌菌株,通过凝胶法初筛和Folk比色法复筛,得到一株产谷氨酰胺转氨酶(transglutaminase,TGase)的菌株,通过形态学、生理生化特征和16S r DNA序列比对证明该菌株是枯草芽孢杆菌(Bacillus subtilis),命名为TGase1318。克隆了该菌株TGase的编码基因tgl,序列长738bp,编码由245个氨基酸组成的蛋白质;TGase的氨基酸序列与NCBI公布的B.subtilis的TGase相似性达94%~100%。用B.subtilis表达载体p TZ和E.coli表达载体p ET21b分别构建了含tgl的重组质粒,转化B.subtilis WB800和E.coli BL21,tgl基因表达产物在70℃下可催化BSA交联,表明tgl在B.subtilis和E.coli中获得了表达且表现出TGase活性,这为其在食品工业上的开发利用打下基础。 

     

    Abstract: By using the dilution plate method, 166 bacterial strains isolated from soil samples and one strain, TGase1318, which could produce transglutaminase, was obtained after screening with gel formation method and Folk's colorimetry method.The strain was identified as Bacillus subtilis according to the morphology, physiological and biochemical characters as well as 16 S r DNA sequence homological analysis.The tgl gene, which was a 738 bp long nucleotide, was cloned from the strain and encoded a 245 residue long TGase.Homological analysis of TGase peptide sequence revealed that it shared 94% 100% conservation with TGase of other B.subtilis strains released by NCBI.The expression recombinant plasmids p TZ- tgl and p ET21b- tgl were constructed and transformed into B.subtilis WB800 and E. coli BL21, respectively. The results of BSA crosslink at 70℃ indicated the tgl gene was expressed in both B.subtilis and E.coli with TGase activity.This study laid a foundation for the application of TGase from B.subtilis TGase1318 to food industry.

     

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