A point mutant in the promoter of transglutaminase gene dramatically increased yield of microbial transglutaminase from Streptomyces mobaraensis TX1

Huang, Yaoxin; Jin, Minfei; Yan, Wenjun; Wu, Qihan; Niu, Yan‐Ning; Zou, Chunjing; Chen, Jia; Chang, Zhongyi; Huang, Jing; Jiang, Deming; Gao, Hongliang

doi:10.1016/j.procbio.2021.11.021

Cited by 6 publications

(4 citation statements)

References 35 publications

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Section: Discussionmentioning

confidence: 63%

“…Notably, compared to the corresponding wild-type strain, the TGase activities of smY2022 and the S. mobaraensis TX1 mutant were increased by 12.2- and 2.15-fold, respectively . Therefore, the transcription enhancement caused by the mutations within the promoter partially contributed to the production of TGase in smY2022.…”

Section: Discussionmentioning

confidence: 95%

“…The same single base mutation was also identified within the TGase promoter of a positive S. mobaraensis TX1 mutant, in which the transcription level of the TGase gene was approximately 50 times higher than that of the wild type. 44 Notably, compared to the corresponding wild-type strain, the TGase activities of smY2022 and the S. mobaraensis TX1 mutant were increased by 12.2-and 2.15-fold, respectively. 44 Therefore, the transcription enhancement caused by the mutations within the promoter partially contributed to the production of TGase in smY2022.…”

Section: Improving the Tgase Production Of S Mobaraensismentioning

confidence: 92%

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Efficient Production of a Thermostable Mutant of Transglutaminase by Streptomyces mobaraensis

Ye,

Yang,

Zhou

et al. 2024

J. Agric. Food Chem.

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The transglutaminase (TGase) from Streptomyces mobaraensis is widely used to improve the texture of protein-based foods. However, wild-type TGase is not heat-resistant, which is unfavorable for its application. In this study, we successfully constructed a S. mobaraensis strain that can efficiently produce TGm2, a thermostable mutant of S. mobaraensis TGase. First, S. mobaraensis DSM40587 was subjected to atmospheric room temperature plasma mutagenesis, generating mutant smY2022 with a 12.2-fold increase in TGase activity. Then, based on the double-crossover recombination, we replaced the coding sequence of the TGase with that of TGm2 in smY2022, obtaining the strain smY2022-TGm2. The extracellular TGase activity of smY2022-TGm2 reached 61.7 U/mL, 147% higher than that of smY2022. Finally, the catalytic properties of TGm2 were characterized. The half-life time at 60 °C and specific activity of TGm2 reached 64 min and 71.15 U/mg, 35.6- and 2.9-fold higher than those of the wild-type TGase, respectively. As indicated by SDS-PAGE analysis, TGm2 exhibited demonstrably better protein cross-linking ability than the wild-type TGase at 70 °C, although both enzymes shared a similar ability at 40 °C. With improved enzyme production and thermal stability, smY2022-TGm2 could be a competitive strain for the industrial production of transglutaminase.

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Section: Discussionmentioning

confidence: 63%

Section: Discussionmentioning

confidence: 95%

Section: Improving the Tgase Production Of S Mobaraensismentioning

confidence: 92%

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Efficient Production of a Thermostable Mutant of Transglutaminase by Streptomyces mobaraensis

Ye,

Yang,

Zhou

et al. 2024

J. Agric. Food Chem.

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“…However, native or wild-type (WT) bacteria for the production of agricultural antibiotics cannot meet the needs of the industry due to the limitations of low yield and long physiological cycle; the increasing requirement for the microbial preparation of antibiotics demands high-yielding microorganism strains applicable to industrial production. Thus far, microbial breeding has been fulfilled by traditional random mutagenesis and screening techniques (Huang et al., 2022 ; Jung et al., 2011 ; Yin et al., 2021 ; Yu et al., 2019a ). Although the performance of such strategies seems fruitful, they are labor and time intensive.…”

Section: Introductionmentioning

confidence: 99%

Improvement of rimocidin production in Streptomyces rimosus M527 by reporter-guided mutation selection

Jiang

Zhang

Huang

et al. 2022

Journal of Industrial Microbiology and Biotechnology

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In this study, we employed a reporter-guided mutation selection (RGMS) strategy to improve the rimocidin production of Streptomyces rimosus M527, which is based on a single-reporter plasmid pAN and atmospheric and room temperature plasma (ARTP). In plasmid pAN, PrimA, a native promoter of the loading module of rimocidin biosynthesis (RimA) was chosen as target, and the kanamycin resistance gene (neo) under the control of PrimA was chosen as reporter gene. The integrative plasmid pAN was introduced into the chromosome of S. rimosus M527 by conjugation to yield the initial strain S. rimosus M527-pAN. Subsequently, mutants of M527-pAN were generated by ARTP. Seventy-nine mutants were obtained in total, of which sixty-seven mutants showed higher level of kanamycin resistance (Kanr) than that of the initial strain M527-pAN. The majority of mutants exhibited a slight increase in rimocidin production compared to M527-pAN. Notably, three mutants, M527-pAN-S34, S38, and S52, which exhibited highest kanamycin resistance among all Kanr mutants, showed 34%, 52% and 45% increase in rimocidin production compared to M527-pAN, respectively. Quantitative RT-PCR analysis revealed that the transcriptional levels of neo and rim genes were increased in mutants M527-pAN-S34, S38, and S52 compared to M527-pAN. These results confirmed that the RGMS approach was successful in improving the rimocidin production in S. rimosus M527.

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Immobilization of transglutaminases and transglutaminase as immobilization agent

Vasić,

Knez,

Leitgeb

2024

Transglutaminase

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A point mutant in the promoter of transglutaminase gene dramatically increased yield of microbial transglutaminase from Streptomyces mobaraensis TX1

Cited by 6 publications

References 35 publications

Efficient Production of a Thermostable Mutant of Transglutaminase by Streptomyces mobaraensis

Efficient Production of a Thermostable Mutant of Transglutaminase by Streptomyces mobaraensis

Improvement of rimocidin production in Streptomyces rimosus M527 by reporter-guided mutation selection

Immobilization of transglutaminases and transglutaminase as immobilization agent

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