Multi-omics Investigation of High-transglutaminase Production Mechanisms in and Co-culture-enhanced Fermentation Strategies

Overview

Journal Front Microbiol

Specialty Microbiology

Date 2025 Feb 20

PMID 39973936

Authors

Huanan Chang

Ziyu Zheng

Hao Li

Yanqiu Xu

Gengyao Zhen

Yao Zhang

Xidong Ren

Xinli Liu

Deqiang Zhu

Affiliations

Soon will be listed here.

Abstract

Transglutaminase (TGase) has been widely applied in the food industry. However, achieving high-yield TGase production remains a challenge, limiting its broader industrial application. In this study, a high-yield strain with stable genetic traits was obtained through UV-ARTP combined mutagenesis, achieving a maximum TGase activity of 13.77 U/mL, representing a 92.43% increase. Using this strain as a forward mutation gene pool, comparative genomic research identified 95 mutated genes, which were mostly due to base substitutions that led to changes in codon usage preference. Transcriptomic analysis revealed significant expression changes in 470 genes, with 232 upregulated and 238 downregulated genes. By investigating potential key regulatory factors, comprehensive analysis indicated that changes in codon usage preference, amino acid metabolism, carbon metabolism, protein export processes, TGase activation, and spore production pathways collectively contributed to the enhancement of TGase activity. Subsequently, the in vitro activation efficiency of TGase was further improved using co-cultivation techniques with neutral proteases secreted by CICC10888, and a TGase activity of 16.91 U/mL was achieved, accounting for a 22.71% increase. This study provides a comprehensive understanding of the mechanisms underlying high-yield TGase production and valuable insights and data references for future research.

References

Anders S, Pyl P, Huber W . HTSeq--a Python framework to work with high-throughput sequencing data. Bioinformatics. 2014; 31(2):166-9. PMC: 4287950. DOI: 10.1093/bioinformatics/btu638. View

Fernie A, Carrari F, Sweetlove L . Respiratory metabolism: glycolysis, the TCA cycle and mitochondrial electron transport. Curr Opin Plant Biol. 2004; 7(3):254-61. DOI: 10.1016/j.pbi.2004.03.007. View

Langmead B, Trapnell C, Pop M, Salzberg S . Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biol. 2009; 10(3):R25. PMC: 2690996. DOI: 10.1186/gb-2009-10-3-r25. View

Zhang L, Zhang L, Han X, Du M, Zhang Y, Feng Z . Enhancement of transglutaminase production in Streptomyces mobaraensis as achieved by treatment with excessive MgCl2. Appl Microbiol Biotechnol. 2011; 93(6):2335-43. DOI: 10.1007/s00253-011-3790-5. View

Oelschlaeger P . Molecular Mechanisms and the Significance of Synonymous Mutations. Biomolecules. 2024; 14(1). PMC: 10813300. DOI: 10.3390/biom14010132. View

Yin X, Rao S, Zhou J, Du G, Chen J, Liu S . Improved Productivity of Transglutaminase by Regulating Zymogen Activation. Front Bioeng Biotechnol. 2022; 10:878795. PMC: 9047793. DOI: 10.3389/fbioe.2022.878795. View

Robinson M, Oshlack A . A scaling normalization method for differential expression analysis of RNA-seq data. Genome Biol. 2010; 11(3):R25. PMC: 2864565. DOI: 10.1186/gb-2010-11-3-r25. View

Robinson M, McCarthy D, Smyth G . edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 2009; 26(1):139-40. PMC: 2796818. DOI: 10.1093/bioinformatics/btp616. View

Stincone A, Prigione A, Cramer T, Wamelink M, Campbell K, Cheung E . The return of metabolism: biochemistry and physiology of the pentose phosphate pathway. Biol Rev Camb Philos Soc. 2014; 90(3):927-63. PMC: 4470864. DOI: 10.1111/brv.12140. View

10.

Kaila V, Wikstrom M . Architecture of bacterial respiratory chains. Nat Rev Microbiol. 2021; 19(5):319-330. DOI: 10.1038/s41579-020-00486-4. View

11.

Angov E, Hillier C, Kincaid R, Lyon J . Heterologous protein expression is enhanced by harmonizing the codon usage frequencies of the target gene with those of the expression host. PLoS One. 2008; 3(5):e2189. PMC: 2364656. DOI: 10.1371/journal.pone.0002189. View

12.

Oteng-Pabi S, Keillor J . Continuous enzyme-coupled assay for microbial transglutaminase activity. Anal Biochem. 2013; 441(2):169-73. DOI: 10.1016/j.ab.2013.07.014. View

13.

Duarte L, Matte C, Bizarro C, Ayub M . Review transglutaminases: part II-industrial applications in food, biotechnology, textiles and leather products. World J Microbiol Biotechnol. 2019; 36(1):11. DOI: 10.1007/s11274-019-2792-9. View

14.

Yang H, Pan L, Lin Y . Purification and on-column activation of a recombinant histidine-tagged pro-transglutaminase after soluble expression in Escherichia coli. Biosci Biotechnol Biochem. 2009; 73(11):2531-4. DOI: 10.1271/bbb.90422. View

15.

Pyrih J, Panek T, Durante I, Raskova V, Cimrhanzlova K, Kriegova E . Vestiges of the Bacterial Signal Recognition Particle-Based Protein Targeting in Mitochondria. Mol Biol Evol. 2021; 38(8):3170-3187. PMC: 8321541. DOI: 10.1093/molbev/msab090. View

16.

Flardh K, Buttner M . Streptomyces morphogenetics: dissecting differentiation in a filamentous bacterium. Nat Rev Microbiol. 2008; 7(1):36-49. DOI: 10.1038/nrmicro1968. View

17.

Liang Z, Qiao J, Li P, Zhang L, Qiao Z, Lin L . A novel Rap-Phr system in Bacillus velezensis NAU-B3 regulates surfactin production and sporulation via interaction with ComA. Appl Microbiol Biotechnol. 2020; 104(23):10059-10074. DOI: 10.1007/s00253-020-10942-z. View

18.

Chen K, Zhang D, Liu S, Wang N, Wang M, Du G . Improvement of transglutaminase production by extending differentiation phase of Streptomyces hygroscopicus: mechanism and application. Appl Microbiol Biotechnol. 2012; 97(17):7711-9. DOI: 10.1007/s00253-012-4614-y. View

19.

Ye J, Yang P, Zhou J, Du G, Liu S . Efficient Production of a Thermostable Mutant of Transglutaminase by . J Agric Food Chem. 2024; 72(8):4207-4216. DOI: 10.1021/acs.jafc.3c07621. View

20.

Holland I, Blight M . ABC-ATPases, adaptable energy generators fuelling transmembrane movement of a variety of molecules in organisms from bacteria to humans. J Mol Biol. 1999; 293(2):381-99. DOI: 10.1006/jmbi.1999.2993. View