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Enhancing transglutaminase production of Streptomyces mobaraensis by iterative mutagenesis breeding with atmospheric and room-temperature plasma (ARTP)

机译:通过大气和室温血浆(ARTP)的迭代诱变育种来增强莫巴拉链霉菌的转谷氨酰胺酶生产

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Objectives: To improve the fermentation production of transglutaminase (TGase) from Streptomyces mobaraensis for applications in the food industry, the atmospheric and room-temperature plasma (ARTP) mutagenesis was applied to breed S. mobaraensis mutants with increased TGase production. Results: After eight rounds of iterative ARTP mutagenesis, four genetically stable mutants, Sm5-V1,Sm6-V13, Sm2-V10, and Sm7-V12, were identified, which showed increased TGase production by 27, 24, 24, and 19%, respectively. The best mutant Sm5-V1 exhibited a maximum TGase activity of 5.85 U/mL during flask fermentation. Compared to the wild-type strain, the transcription levels of the zymogen TGase genes in the mutants increased significantly as indicated by quantitative real-time PCR, while the gene nucleotide sequences of the mutants did not change at all. It was shown that the overexpression of TGase zymogen gene in the mutants contributes to the increase in TGase production. Conclusions: ARTP is a potentially efficient tool for microbial mutation breeding to bring some significant changes required for the industrial applications.
机译:目的:为提高莫巴拉链霉菌的转谷氨酰胺酶(TGase)的发酵生产,以用于食品工业,利用常压和室温血浆(ARTP)诱变技术,培育出TGase产量增加的莫氏沙门氏菌突变体。结果:经过八轮迭代ARTP诱变,鉴定了四个遗传稳定的突变体Sm5-V1,Sm6-V13,Sm2-V10和Sm7-V12,显示TGase产量增加了27%,24%,24%和19% , 分别。最佳的Sm5-V1突变体在烧瓶发酵过程中表现出最大的TGase活性为5.85 U / mL。与野生型菌株相比,突变体中酶原TGase基因的转录水平显着增加,如实时定量PCR所示,而突变体的基因核苷酸序列则完全没有变化。结果表明,突变体中TGase酶原基因的过表达促进了TGase产量的增加。结论:ARTP是微生物突变育种的潜在有效工具,可带来工业应用所需的一些重大变化。

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