关键调控因子增强茂原链霉菌转谷氨酰胺酶转录表达的作用
Title: Effect of key regulators in augmenting transcriptional expression of Transglutaminase in Streptomyces mobaraensis
Download website:www.elsevier.com/locate/biortech
Abstract: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 MTGase biosynthesis 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, MgCl2, 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.
Methods：
Results:
培养基添加剂 | 蛋白酶 (20 IU) | MgCl2 (0.2 M) | CTAB (1.0%) | All in one | PMSF | NaF | bleomycin sulphate |
相对表达量 | 2.86% | 1.66% | 2.48% | 4.27% | -14.09% | -7.52% | -8.21% |
Conclusion：
CTAB、MgCl2以及蛋白酶显著提升了MTGase的活性，且这三种关键调节剂表现出明显的协同作用。
CTAB、MgCl2以及蛋白酶显著上调MTGase转录表达，而硫酸博来霉素、PMSF以及NaF从转录水平上明显抑制MTGase的表达。
Contact: Feng Yuxuan
E-mail: 1032368607@qq.com