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Combinatorial engineering for efficient production of protein-glutaminase in Bacillus subtilis.
Abstract
Protein-glutaminase (EC 3.5.1.44, PG) converts protein glutamine residues in proteins and peptides into glutamic acid residue, and markedly improves the solubility, emulsification, and foaming properties of food proteins. However, the source bacteria, Chryseobacterium proteolyticum, have low enzyme production ability, inefficient genetic operation, and high production cost. Therefore, it is critical to establish an efficient expression system for active PG. Here, combinatorial engineering was developed for high-yield production of PG in Bacillus subtilis. First, we evaluated different B. subtilis strains for PG self-activation. Then, combinatorial optimization involving promoters, signal peptides, and culture medium was applied to produce active recombinant PG in a B. subtilis expression system. Through combinatorial engineering, PG enzyme activity reached 3.23 U/mL in shaken-flask cultures. Active PG with the yield of 7.07 U/mL was obtained at 40 h by the P SecA-YdeJ combination in fed-batch fermentation, which is the highest yield of PG in existing reports.
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“During fermentation, B. subtilis may produce extracellular proteases that can self-activate protein glutaminase, which is released in the inactive pro-enzyme form and can facilitate efficient substrate utilization by the bacterium [31].”
From Optimization of Bacillus subtilis PW12 biomass production using RSM: a preliminary study towards single-cell protein production for aquaculture (2024) · cites this paper as [31]
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Cites
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Cited by
Application Prospect of Protein-Glutaminase in the Development of Plant-Based Protein Foods
Liu, X., Wang, C., Zhang, X., Zhang, G., Zhou, J., & Jian, C. (2022) · Foods · doi:10.3390/foods11030440 · Cited by 67
Self-citationProtein-glutaminase: Research progress and prospect in food manufacturing
Zhang, G., Ma, S., Liu, X., Yin, X., Liu, S., Zhou, J., & Du, G. (2021) · Food Bioscience · doi:10.1016/j.fbio.2021.101314 · Cited by 58
Self-citationZheng, N., Long, M., Zhang, Z., Osire, T., Zan, Q., Zhou, H., Du, S., & Xia, X. (2023) · Food Hydrocolloids · doi:10.1016/j.foodhyd.2023.108594 · Cited by 46
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Metadata
- Authors
- Yin, X., Zhang, G., Zhou, J., Li, J., & Du, G.
- Affiliations
- Jiangnan University (CN)
- Corresponding author
- Zhang, Guoqiang
- Du, Guocheng
- Venue
- Enzyme and Microbial Technology
- Publisher
- Elsevier BV
- Published
- 2021-10
- Volume
- 150
- Pages
- 109863
- ISSN
- 0141-0229 (ISSN-L)
- OpenAlex
- W3175985869
- Cited by
- 37
- References
- 25
- Citations by year
- 2026: 5 · 2025: 9 · 2024: 11 · 2023: 6 · 2022: 5 · 2021: 1
- FWCI
- 3.01
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Enzyme and Microbial Technology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Keywords
- protein-glutaminase, Bacillus subtilis, recombinant enzyme expression, combinatorial engineering, promoter optimization, signal peptide, culture medium optimization, fed-batch fermentation
- MeSH terms
- Bacillus subtilis, Bacterial Proteins, Fermentation, Glutaminase, Protein Engineering, Protein Sorting Signals, Chryseobacterium
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- Yin, X., Zhang, G., Zhou, J., Li, J., & Du, G. (2021). Combinatorial engineering for efficient production of protein-glutaminase in Bacillus subtilis. Enzyme and Microbial Technology. https://doi.org/10.1016/j.enzmictec.2021.109863
