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http://dx.doi.org/10.4014/jmb.2002.02049

Heterologous Expression of Recombinant Transglutaminase in Bacillus subtilis SCK6 with Optimized Signal Peptide and Codon, and Its Impact on Gelatin Properties  

Wang, Shiting (Key Laboratory of Leather Chemistry and Engineering (Sichuan University), Ministry of Education)
Yang, Zhigang (Chengdu Jinkai Bioengineering Co., Ltd.)
Li, Zhenjiang (Chengdu Jinkai Bioengineering Co., Ltd.)
Tian, Yongqiang (Key Laboratory of Leather Chemistry and Engineering (Sichuan University), Ministry of Education)
Publication Information
Journal of Microbiology and Biotechnology / v.30, no.7, 2020 , pp. 1082-1091 More about this Journal
Abstract
Microbial transglutaminases (MTGs) are widely used in the food industry. In this study, the MTG gene of Streptomyces sp. TYQ1024 was cloned and expressed in a food-grade bacterial strain, Bacillus subtilis SCK6. Extracellular activity of the MTG after codon and signal peptide (SP Ync M) optimization was 20 times that of the pre-optimized enzyme. After purification, the molecular weight of the MTG was 38 kDa and the specific activity was 63.75 U/mg. The optimal temperature and pH for the recombinant MTG activity were 50℃ and 8.0, respectively. MTG activity increased 1.42-fold in the presence of β-ME and 1.6-fold in the presence of DTT. Moreover, 18% sodium chloride still resulted in 83% enzyme activity, which showed good salt tolerance. Cross-linking gelatin with the MTG increased the strength of gelatin 1.67 times and increased the thermal denaturation temperature from 61.8 to 75.8℃. The MTG also significantly increased the strength and thermal stability of gelatin. These characteristics demonstrated the huge commercial potential of MTG, such as for applications in salted protein foods.
Keywords
MTG; heterologous expression; signal peptide optimization; Bacillus subtilis SCK6; enzymatic properties;
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