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Enhanced Production of Transglutaminase in Streptomyces mobaraensis through Random Mutagenesis and Site-Directed Genetic Modification
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2021-03-02 , DOI: 10.1021/acs.jafc.1c00645 Xiaoqiang Yin 1, 2 , Yangyang Li 1, 2 , Jingwen Zhou 1, 2, 3 , Shengqi Rao 4 , Guocheng Du 2, 5 , Jian Chen 1, 2, 3 , Song Liu 1, 2
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2021-03-02 , DOI: 10.1021/acs.jafc.1c00645 Xiaoqiang Yin 1, 2 , Yangyang Li 1, 2 , Jingwen Zhou 1, 2, 3 , Shengqi Rao 4 , Guocheng Du 2, 5 , Jian Chen 1, 2, 3 , Song Liu 1, 2
Affiliation
Streptomyces transglutaminase (TGase) is widely used to improve food texture properties. In this study, random mutagenesis and site-directed genetic modification were used to improve the production of TGase in Streptomyces mobaraensis. First, S. mobaraensis DSM40587 (smWT) was subjected to atmospheric and room-temperature plasma mutagenesis, and then a mutant (smY2019) with a 5.5-fold increase in TGase yield was screened from approximately 3000 × 25 (round) mutants. Compared to smWT, smY2019 exhibits a 3.2-fold higher TGase mRNA level and two site mutations within the −10 region of the TGase promoter. The recombinant expression analysis in the TGase-deficient S. mobaraensis suggests that the mutated TGase promoter is more robust than the wild-type one. Finally, we integrated two additional TGase expression cassettes into the smY2019 genome, yielding the recombinant strain smY2019-3C with a 103% increase in TGase production compared to smY2019. The smY2019-3C strain with 40 U/mL of TGase yield could be a suitable candidate for the industrial production of TGase.
中文翻译:
通过随机诱变和定点遗传修饰提高莫巴链霉菌转谷氨酰胺酶的产量
链霉菌转谷氨酰胺酶(TGase)被广泛用于改善食物质地。在这项研究中,随机诱变和定点遗传修饰被用来提高莫原链霉菌中TGase的产生。首先,对mobaraensis DSM40587(smWT)进行大气和室温血浆诱变,然后从大约3000×25(圆形)突变体中筛选出TGase产量增加5.5倍的突变体(smY2019)。与smWT相比,smY2019的TGase mRNA水平高3.2倍,并且在TGase启动子的-10位区域内有两个位点突变。TGase缺陷型桑原酵母的重组表达分析提示突变的TGase启动子比野生型更强。最后,我们将另外两个TGase表达盒整合到smY2019基因组中,产生了重组菌株smY2019-3C,与smY2019相比,TGase产量增加了103%。TGase产量为40 U / mL的smY2019-3C菌株可能适合工业生产TGase。
更新日期:2021-03-17
中文翻译:
通过随机诱变和定点遗传修饰提高莫巴链霉菌转谷氨酰胺酶的产量
链霉菌转谷氨酰胺酶(TGase)被广泛用于改善食物质地。在这项研究中,随机诱变和定点遗传修饰被用来提高莫原链霉菌中TGase的产生。首先,对mobaraensis DSM40587(smWT)进行大气和室温血浆诱变,然后从大约3000×25(圆形)突变体中筛选出TGase产量增加5.5倍的突变体(smY2019)。与smWT相比,smY2019的TGase mRNA水平高3.2倍,并且在TGase启动子的-10位区域内有两个位点突变。TGase缺陷型桑原酵母的重组表达分析提示突变的TGase启动子比野生型更强。最后,我们将另外两个TGase表达盒整合到smY2019基因组中,产生了重组菌株smY2019-3C,与smY2019相比,TGase产量增加了103%。TGase产量为40 U / mL的smY2019-3C菌株可能适合工业生产TGase。