Enzyme and Microbial Biochemistry Laboratory, Department of Chemistry, Indian Institute of Technology Delhi, New Delhi 110016, India.
Enzyme and Microbial Biochemistry Laboratory, Department of Chemistry, Indian Institute of Technology Delhi, New Delhi 110016, India.
Bioresour Technol. 2021 Nov;340:125627. doi: 10.1016/j.biortech.2021.125627. Epub 2021 Jul 22.
Transglutaminase forms isopeptide bonds in proteins which are helpful in various industrial applications. However, low productivity and high cost are the major bottlenecks for industrial Transglutaminase production. The present study describes the regulatory mechanism of microbial Transglutaminase (MTGase) biosynthesis from Streptomyces mobaraensis and the effect of key regulators to maximize production. The transcriptional responses under the effect of various key modulators of MTGasebiosynthesis were evaluated. Productivity of MTGase with novel biosynthesis approach by regulators augmentation was correlated by transcriptional profiling. The optimization by key modulators by combinational supplementation led to 2-fold rise in activity. The functional attributes, the copy number of MTGase gene and relative changes were assessed by Real-Time quantitative PCR. Protease, MgCl, CTAB induced upregulation, whereas PMSF, NaF and bleomycin sulphate showed inhibitory action on MTGase production and activity. The optimization by combinational supplementation of key modulators led to 4.27-fold increase (6.11 IU/mL) in production.
转谷氨酰胺酶在蛋白质中形成异肽键,这在各种工业应用中很有帮助。然而,低产量和高成本是工业转谷氨酰胺酶生产的主要瓶颈。本研究描述了来自藤黄微球菌的微生物转谷氨酰胺酶(MTGase)生物合成的调控机制,以及关键调节剂对最大限度提高产量的影响。评估了 MTGase 生物合成受各种关键调节剂影响下的转录响应。通过转录谱分析,将新型生物合成方法的 MTGase 生产力与调控因子的增加相关联。通过组合补充关键调节剂进行优化,使活性提高了 2 倍。通过实时定量 PCR 评估功能属性、MTGase 基因的拷贝数和相对变化。蛋白酶、MgCl、CTAB 诱导上调,而 PMSF、NaF 和博来霉素硫酸盐对 MTGase 的产生和活性表现出抑制作用。通过组合补充关键调节剂的优化,使产量提高了 4.27 倍(6.11 IU/mL)。