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Extracellular Production Of Active-Form Streptomyces Mobaraensis Transglutaminase In Bacillus Subtilis

文献类型： 外文期刊

作者： Fu, LH; Wang, Y; Ju, JS; Cheng, L; Xu, YQ; Yu, B; Wang, LM

作者机构：

关键词： Transglutaminase (Tg); Streptomyces; Bacillus Subtilis; Intein

期刊名称： APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

ISSN： 0175-7598

年卷期： 2020 年 104 卷 2 期

页码：

收录情况： JCR（2021版） ; EI（2021版）

摘要： Transglutaminase (TG) from Streptomyces mobaraensis has been widely used in the food industry. It is secreted naturally as an inactive zymogen, which is then activated by the removal of the N-terminal pro-peptide. In this study, the mtg gene from S. mobaraensis was expressed in a food-grade strain of bacterium, Bacillus subtilis. When its native signal peptide was replaced by signal peptide SacB (SPsacB) and the pro-peptide was replaced by that derived from S. hygroscopicus, an extracellular activity of 16.1 U/mg was observed. A modified Saccharomyces cerevisiae vacuolar ATPase subunit (VMA) intein was introduced into the zymogen to simplify its activation process by controlling temperature. When the cleavage site in the C-terminal of VMA was placed between the pro-peptide and core domain, the activation process was carried out at 18 degrees C. Promoter replacement further increased the enzymatic activity. Finally, the extracellular enzymatic activity reached 2.6 U/mg under the control of the constitutive promoter P-yvyD. This is the first report on the extracellular production of active-form Streptomyces TG in B. subtilis without splicing with the cleavage enzyme.

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