Biofilm-Based Immobilization Fermentation for Continuous hEGF Production in <i>Saccharomyces cerevisiae</i>
Biofilms can enhance industrial fermentation efficiency by increasing cell density, stability, and metabolic activity and have been successfully applied to the continuous production of many small-molecule chemicals. However, the continuous production of proteins by biofilms has been less studied. Th...
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| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-12-01
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| Series: | Fermentation |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2311-5637/10/12/661 |
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| Summary: | Biofilms can enhance industrial fermentation efficiency by increasing cell density, stability, and metabolic activity and have been successfully applied to the continuous production of many small-molecule chemicals. However, the continuous production of proteins by biofilms has been less studied. This study used secretory human epidermal growth factor (hEGF) as a representative product to evaluate and optimize biofilm-based continuous protein production. First, by deleting the protease and overexpressing eight key genes involved in protein secretion in <i>Saccharomyces cerevisiae</i>, the yield of hEGF was improved by 82.6% from 77.4 to 141.3 mg/L in shake flasks. Subsequently, the flocculation genes <i>FLO11</i> and <i>ALS3</i> were introduced to facilitate the establishment of a biofilm-based continuous immobilization fermentation model. The optimal strain SIC-<i>ALS3</i>-<i>PDI1</i> produced 583.8 mg/L of hEGF, with a productivity of 4.9 mg/L/h during traditional free-cell fermentation, while it produced an average of 300.0 mg/L of hEGF in 10 continuous batches of biofilm-based fermentation, with a productivity of 6.3 mg/L/h. Although the hEGF production in biofilms was lower than that in free-cell fermentation, biofilm fermentation demonstrated greater productivity, with the advantage of not requiring seed culture for each batch of fermentation. This study provided a valuable reference for the biofilm-based production of other peptides. |
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| ISSN: | 2311-5637 |