Cargando…
Synthetic propeptide design to enhance the secretion of heterologous proteins by Saccharomyces cerevisiae
Heterologous protein production in Saccharomyces cerevisiae is a useful and effective strategy with many advantages, including the secretion of proteins that require posttranslational processing. However, heterologous proteins in S. cerevisiae are often secreted at comparatively low levels. To impro...
Autores principales: | , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9178654/ https://www.ncbi.nlm.nih.gov/pubmed/35765186 http://dx.doi.org/10.1002/mbo3.1300 |
_version_ | 1784723102467358720 |
---|---|
author | Cho, Ji Sung Oh, Hye Ji Jang, Young Eun Kim, Hyun Jin Kim, Areum Song, Jong‐Am Lee, Eun Jung Lee, Jeewon |
author_facet | Cho, Ji Sung Oh, Hye Ji Jang, Young Eun Kim, Hyun Jin Kim, Areum Song, Jong‐Am Lee, Eun Jung Lee, Jeewon |
author_sort | Cho, Ji Sung |
collection | PubMed |
description | Heterologous protein production in Saccharomyces cerevisiae is a useful and effective strategy with many advantages, including the secretion of proteins that require posttranslational processing. However, heterologous proteins in S. cerevisiae are often secreted at comparatively low levels. To improve the production of the heterologous protein, human granulocyte colony‐stimulating factor (hG‐CSF) in S. cerevisiae, a secretion‐enhancing peptide cassette including an hIL‐1β‐derived propeptide, was added and used as a secretion enhancer to alleviate specific bottlenecks in the yeast secretory pathway. The effects of three key parameters—N‐glycosylation, net negative charge balance, and glycine‐rich flexible linker—were investigated in batch cultures of S. cerevisiae. Using a three‐stage design involving screening, selection, and optimization, the production and secretion of hG‐CSF by S. cerevisiae were significantly increased. The amount of extracellular mature hG‐CSF produced by the optimized propeptide after the final stage increased by 190% compared to that of the original propeptide. Although hG‐CSF was used as the model protein in the current study, this strategy applies to the enhanced production of other heterologous proteins, using S. cerevisiae as the host. |
format | Online Article Text |
id | pubmed-9178654 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-91786542022-06-13 Synthetic propeptide design to enhance the secretion of heterologous proteins by Saccharomyces cerevisiae Cho, Ji Sung Oh, Hye Ji Jang, Young Eun Kim, Hyun Jin Kim, Areum Song, Jong‐Am Lee, Eun Jung Lee, Jeewon Microbiologyopen Original Articles Heterologous protein production in Saccharomyces cerevisiae is a useful and effective strategy with many advantages, including the secretion of proteins that require posttranslational processing. However, heterologous proteins in S. cerevisiae are often secreted at comparatively low levels. To improve the production of the heterologous protein, human granulocyte colony‐stimulating factor (hG‐CSF) in S. cerevisiae, a secretion‐enhancing peptide cassette including an hIL‐1β‐derived propeptide, was added and used as a secretion enhancer to alleviate specific bottlenecks in the yeast secretory pathway. The effects of three key parameters—N‐glycosylation, net negative charge balance, and glycine‐rich flexible linker—were investigated in batch cultures of S. cerevisiae. Using a three‐stage design involving screening, selection, and optimization, the production and secretion of hG‐CSF by S. cerevisiae were significantly increased. The amount of extracellular mature hG‐CSF produced by the optimized propeptide after the final stage increased by 190% compared to that of the original propeptide. Although hG‐CSF was used as the model protein in the current study, this strategy applies to the enhanced production of other heterologous proteins, using S. cerevisiae as the host. John Wiley and Sons Inc. 2022-06-09 /pmc/articles/PMC9178654/ /pubmed/35765186 http://dx.doi.org/10.1002/mbo3.1300 Text en © 2022 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Articles Cho, Ji Sung Oh, Hye Ji Jang, Young Eun Kim, Hyun Jin Kim, Areum Song, Jong‐Am Lee, Eun Jung Lee, Jeewon Synthetic propeptide design to enhance the secretion of heterologous proteins by Saccharomyces cerevisiae |
title | Synthetic propeptide design to enhance the secretion of heterologous proteins by Saccharomyces cerevisiae
|
title_full | Synthetic propeptide design to enhance the secretion of heterologous proteins by Saccharomyces cerevisiae
|
title_fullStr | Synthetic propeptide design to enhance the secretion of heterologous proteins by Saccharomyces cerevisiae
|
title_full_unstemmed | Synthetic propeptide design to enhance the secretion of heterologous proteins by Saccharomyces cerevisiae
|
title_short | Synthetic propeptide design to enhance the secretion of heterologous proteins by Saccharomyces cerevisiae
|
title_sort | synthetic propeptide design to enhance the secretion of heterologous proteins by saccharomyces cerevisiae |
topic | Original Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9178654/ https://www.ncbi.nlm.nih.gov/pubmed/35765186 http://dx.doi.org/10.1002/mbo3.1300 |
work_keys_str_mv | AT chojisung syntheticpropeptidedesigntoenhancethesecretionofheterologousproteinsbysaccharomycescerevisiae AT ohhyeji syntheticpropeptidedesigntoenhancethesecretionofheterologousproteinsbysaccharomycescerevisiae AT jangyoungeun syntheticpropeptidedesigntoenhancethesecretionofheterologousproteinsbysaccharomycescerevisiae AT kimhyunjin syntheticpropeptidedesigntoenhancethesecretionofheterologousproteinsbysaccharomycescerevisiae AT kimareum syntheticpropeptidedesigntoenhancethesecretionofheterologousproteinsbysaccharomycescerevisiae AT songjongam syntheticpropeptidedesigntoenhancethesecretionofheterologousproteinsbysaccharomycescerevisiae AT leeeunjung syntheticpropeptidedesigntoenhancethesecretionofheterologousproteinsbysaccharomycescerevisiae AT leejeewon syntheticpropeptidedesigntoenhancethesecretionofheterologousproteinsbysaccharomycescerevisiae |