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...
Saved in:
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2024-12-01
|
| Series: | Fermentation |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2311-5637/10/12/661 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850050302716149760 |
|---|---|
| author | Kaiqi Zhi Zhiguo An Mingyang Zhang Kehan Liu Yafan Cai Zhenyu Wang Di Zhang Jinle Liu Zhi Wang Chenjie Zhu Dong Liu Sheng Yang Hanjie Ying |
| author_facet | Kaiqi Zhi Zhiguo An Mingyang Zhang Kehan Liu Yafan Cai Zhenyu Wang Di Zhang Jinle Liu Zhi Wang Chenjie Zhu Dong Liu Sheng Yang Hanjie Ying |
| author_sort | Kaiqi Zhi |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-ec116f8d4d834f89b889e54c1a9b1d2a |
| institution | DOAJ |
| issn | 2311-5637 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj-art-ec116f8d4d834f89b889e54c1a9b1d2a2025-08-20T02:53:30ZengMDPI AGFermentation2311-56372024-12-01101266110.3390/fermentation10120661Biofilm-Based Immobilization Fermentation for Continuous hEGF Production in <i>Saccharomyces cerevisiae</i>Kaiqi Zhi0Zhiguo An1Mingyang Zhang2Kehan Liu3Yafan Cai4Zhenyu Wang5Di Zhang6Jinle Liu7Zhi Wang8Chenjie Zhu9Dong Liu10Sheng Yang11Hanjie Ying12School of Chemical Engineering Zhengzhou University, Zhengzhou 450001, ChinaSchool of Chemical Engineering Zhengzhou University, Zhengzhou 450001, ChinaSchool of Chemical Engineering Zhengzhou University, Zhengzhou 450001, ChinaSchool of Chemical Engineering Zhengzhou University, Zhengzhou 450001, ChinaSchool of Chemical Engineering Zhengzhou University, Zhengzhou 450001, ChinaNational Engineering Research Center for Biotechnology, Nanjing Tech University, Nanjing 210009, ChinaNational Engineering Research Center for Biotechnology, Nanjing Tech University, Nanjing 210009, ChinaSchool of Chemical Engineering Zhengzhou University, Zhengzhou 450001, ChinaSchool of Chemical Engineering Zhengzhou University, Zhengzhou 450001, ChinaNational Engineering Research Center for Biotechnology, Nanjing Tech University, Nanjing 210009, ChinaNational Engineering Research Center for Biotechnology, Nanjing Tech University, Nanjing 210009, ChinaShanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 300 Fenglin Road, Shanghai 200032, ChinaSchool of Chemical Engineering Zhengzhou University, Zhengzhou 450001, ChinaBiofilms 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.https://www.mdpi.com/2311-5637/10/12/661hEGF<i>S. cerevisiae</i>biofilmimmobilization fermentation |
| spellingShingle | Kaiqi Zhi Zhiguo An Mingyang Zhang Kehan Liu Yafan Cai Zhenyu Wang Di Zhang Jinle Liu Zhi Wang Chenjie Zhu Dong Liu Sheng Yang Hanjie Ying Biofilm-Based Immobilization Fermentation for Continuous hEGF Production in <i>Saccharomyces cerevisiae</i> Fermentation hEGF <i>S. cerevisiae</i> biofilm immobilization fermentation |
| title | Biofilm-Based Immobilization Fermentation for Continuous hEGF Production in <i>Saccharomyces cerevisiae</i> |
| title_full | Biofilm-Based Immobilization Fermentation for Continuous hEGF Production in <i>Saccharomyces cerevisiae</i> |
| title_fullStr | Biofilm-Based Immobilization Fermentation for Continuous hEGF Production in <i>Saccharomyces cerevisiae</i> |
| title_full_unstemmed | Biofilm-Based Immobilization Fermentation for Continuous hEGF Production in <i>Saccharomyces cerevisiae</i> |
| title_short | Biofilm-Based Immobilization Fermentation for Continuous hEGF Production in <i>Saccharomyces cerevisiae</i> |
| title_sort | biofilm based immobilization fermentation for continuous hegf production in i saccharomyces cerevisiae i |
| topic | hEGF <i>S. cerevisiae</i> biofilm immobilization fermentation |
| url | https://www.mdpi.com/2311-5637/10/12/661 |
| work_keys_str_mv | AT kaiqizhi biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT zhiguoan biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT mingyangzhang biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT kehanliu biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT yafancai biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT zhenyuwang biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT dizhang biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT jinleliu biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT zhiwang biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT chenjiezhu biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT dongliu biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT shengyang biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei AT hanjieying biofilmbasedimmobilizationfermentationforcontinuoushegfproductioninisaccharomycescerevisiaei |