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Article
2025
Recombinant production of bovine αS1-casein in genome-reduced Bacillus subtilis strain IIG-Bs-20-5-1
Recombinant production of bovine αS1-casein in genome-reduced Bacillus subtilis strain IIG-Bs-20-5-1
Abstract
Background: Cow’s milk represents an important protein source. Here, especially casein proteins are important components, which might be a promising source of alternative protein production by microbial expression systems. Nevertheless, caseins are difficult-to-produce proteins, making heterologous production challenging. However, the potential of genome-reduced Bacillus subtilis was applied for the recombinant production of bovine αS1-casein protein. Methods: A plasmid-based gene expression system was established in B. subtilis allowing the production of his-tagged codon-optimized bovine αS1-casein. Upscaling in a fed-batch bioreactor system for high cell-density fermentation processes allowed for efficient recombinant αS1-casein production. After increasing the molecular abundance of the recombinant αS1-casein protein using immobilized metal affinity chromatography, zeta potential and particle size distribution were determined in comparison to native bovine αS1-casein. Results: Non-sporulating B. subtilis strain BMV9 and genome-reduced B. subtilis strain IIG-Bs-20-5-1 were applied for recombinant αS1-casein production. Casein was detectable only in the insoluble protein fraction of the genome-reduced B. subtilis strain. Subsequent high cell-density fed-batch bioreactor cultivations using strain IIG-Bs-20-5-1 resulted in a volumetric casein titer of 56.9 mg/L and a yield of 1.6 mgcasein/gCDW after reducing the B. subtilis protein content. Comparative analyses of zeta potential and particle size between pre-cleaned recombinant and native αS1-casein showed pH-mediated differences in aggregation behavior. Conclusions: The study demonstrates the potential of B. subtilis for the recombinant production of bovine αS1-casein and underlines the potential of genome reduction for the bioproduction of difficult-to-produce proteins.
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Microorganisms, 13 (2025), 1, 60.
https://doi.org/10.3390/microorganisms13010060.
ISSN: 2076-2607
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English
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660 Chemical engineerin
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Sustainable Development Goals
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@article{Biermann2025,
doi = {10.3390/microorganisms13010060},
author = {Biermann, Lennart and Tadele, Lea Rahel and Benatto Perino, Elvio Henrique et al.},
title = {Recombinant production of bovine αS1-casein in genome-reduced Bacillus subtilis strain IIG-Bs-20-5-1},
journal = {Microorganisms},
year = {2025},
volume = {13},
number = {1},
}