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...

Descripción completa

Detalles Bibliográficos
Autores principales: Cho, Ji Sung, Oh, Hye Ji, Jang, Young Eun, Kim, Hyun Jin, Kim, Areum, Song, Jong‐Am, Lee, Eun Jung, Lee, Jeewon
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