Functional expression of recombinant insulins in Saccharomyces cerevisiae
Abstract Background Since 1982, recombinant insulin has been used as a substitute for pancreatic insulin from animals. However, increasing demand in medical and food industries warrants the development of more efficient production methods. In this study, we aimed to develop a novel and efficient met...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-11-01
|
| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-024-02571-2 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1846164796347514880 |
|---|---|
| author | Mi-Jin Kim Se-Lin Park Hyun-Jin Kim Bong Hyun Sung Jung-Hoon Sohn Jung-Hoon Bae |
| author_facet | Mi-Jin Kim Se-Lin Park Hyun-Jin Kim Bong Hyun Sung Jung-Hoon Sohn Jung-Hoon Bae |
| author_sort | Mi-Jin Kim |
| collection | DOAJ |
| description | Abstract Background Since 1982, recombinant insulin has been used as a substitute for pancreatic insulin from animals. However, increasing demand in medical and food industries warrants the development of more efficient production methods. In this study, we aimed to develop a novel and efficient method for insulin production using a yeast secretion system. Methods Here, insulin C-peptide was replaced with a hydrophilic fusion partner (HL18) containing an affinity tag for the hypersecretion and easy purification of proinsulin. The HL18 fusion partner was then removed by in vitro processing with the Kex2 endoprotease (Kex2p), and authentic insulin was recovered via affinity chromatography. To improve the insulin functions, molecular chaperones of the host strain were reinforced via the constitutive expression of HAC1. Results The developed method was successfully applied for the expression of cow, pig, and chicken insulins in yeast. Moreover, biological activity of recombinant insulins was confirmed by growth stimulation of cell line. Conclusions Therefore, replacement of the C-peptide of insulin with the HL18 fusion partner and use of Kex2p for in vitro processing of proinsulin guarantees the economic production of animal insulins in yeast. |
| format | Article |
| id | doaj-art-e50b645f795f495f8eab0c6bed2f0fcb |
| institution | Kabale University |
| issn | 1475-2859 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj-art-e50b645f795f495f8eab0c6bed2f0fcb2024-11-17T12:55:11ZengBMCMicrobial Cell Factories1475-28592024-11-012311910.1186/s12934-024-02571-2Functional expression of recombinant insulins in Saccharomyces cerevisiaeMi-Jin Kim0Se-Lin Park1Hyun-Jin Kim2Bong Hyun Sung3Jung-Hoon Sohn4Jung-Hoon Bae5Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)Cellapy Bio Inc. Bio-Venture CenterSynthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)Abstract Background Since 1982, recombinant insulin has been used as a substitute for pancreatic insulin from animals. However, increasing demand in medical and food industries warrants the development of more efficient production methods. In this study, we aimed to develop a novel and efficient method for insulin production using a yeast secretion system. Methods Here, insulin C-peptide was replaced with a hydrophilic fusion partner (HL18) containing an affinity tag for the hypersecretion and easy purification of proinsulin. The HL18 fusion partner was then removed by in vitro processing with the Kex2 endoprotease (Kex2p), and authentic insulin was recovered via affinity chromatography. To improve the insulin functions, molecular chaperones of the host strain were reinforced via the constitutive expression of HAC1. Results The developed method was successfully applied for the expression of cow, pig, and chicken insulins in yeast. Moreover, biological activity of recombinant insulins was confirmed by growth stimulation of cell line. Conclusions Therefore, replacement of the C-peptide of insulin with the HL18 fusion partner and use of Kex2p for in vitro processing of proinsulin guarantees the economic production of animal insulins in yeast.https://doi.org/10.1186/s12934-024-02571-2InsulinSecretionKex2pCultured meatSaccharomyces cerevisiae |
| spellingShingle | Mi-Jin Kim Se-Lin Park Hyun-Jin Kim Bong Hyun Sung Jung-Hoon Sohn Jung-Hoon Bae Functional expression of recombinant insulins in Saccharomyces cerevisiae Microbial Cell Factories Insulin Secretion Kex2p Cultured meat Saccharomyces cerevisiae |
| title | Functional expression of recombinant insulins in Saccharomyces cerevisiae |
| title_full | Functional expression of recombinant insulins in Saccharomyces cerevisiae |
| title_fullStr | Functional expression of recombinant insulins in Saccharomyces cerevisiae |
| title_full_unstemmed | Functional expression of recombinant insulins in Saccharomyces cerevisiae |
| title_short | Functional expression of recombinant insulins in Saccharomyces cerevisiae |
| title_sort | functional expression of recombinant insulins in saccharomyces cerevisiae |
| topic | Insulin Secretion Kex2p Cultured meat Saccharomyces cerevisiae |
| url | https://doi.org/10.1186/s12934-024-02571-2 |
| work_keys_str_mv | AT mijinkim functionalexpressionofrecombinantinsulinsinsaccharomycescerevisiae AT selinpark functionalexpressionofrecombinantinsulinsinsaccharomycescerevisiae AT hyunjinkim functionalexpressionofrecombinantinsulinsinsaccharomycescerevisiae AT bonghyunsung functionalexpressionofrecombinantinsulinsinsaccharomycescerevisiae AT junghoonsohn functionalexpressionofrecombinantinsulinsinsaccharomycescerevisiae AT junghoonbae functionalexpressionofrecombinantinsulinsinsaccharomycescerevisiae |