Construction and screening of a glycosylphosphatidylinositol protein deletion library in Pichia pastoris
BACKGROUND: Glycosylphosphatidylinositol (GPI)-anchored glycoproteins have diverse intrinsic functions in yeasts, and they also have different uses in vitro. In this study, the functions of potential GPI proteins in Pichia pastoris were explored by gene knockout approaches. RESULTS: Through an exten...
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ftpubmed:oai:pubmedcentral.nih.gov:7446130 2023-05-15T14:04:46+02:00 Construction and screening of a glycosylphosphatidylinositol protein deletion library in Pichia pastoris Wang, Pan Lin, Ying Zou, Chengjuan Zhao, Fengguang Liang, Shuli Zheng, Suiping Han, Shuangyan 2020-08-24 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7446130/ http://www.ncbi.nlm.nih.gov/pubmed/32838766 https://doi.org/10.1186/s12866-020-01928-y en eng BioMed Central http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7446130/ http://www.ncbi.nlm.nih.gov/pubmed/32838766 http://dx.doi.org/10.1186/s12866-020-01928-y © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. CC0 PDM CC-BY BMC Microbiol Research Article Text 2020 ftpubmed https://doi.org/10.1186/s12866-020-01928-y 2020-08-30T00:45:41Z BACKGROUND: Glycosylphosphatidylinositol (GPI)-anchored glycoproteins have diverse intrinsic functions in yeasts, and they also have different uses in vitro. In this study, the functions of potential GPI proteins in Pichia pastoris were explored by gene knockout approaches. RESULTS: Through an extensive knockout of GPI proteins in P. pastoris, a single-gene deletion library was constructed for 45 predicted GPI proteins. The knockout of proteins may lead to the activation of a cellular response named the ‘compensatory mechanism’, which is characterized by changes in the content and relationship between cell wall polysaccharides and surface proteins. Among the 45 deletion strains, five showed obvious methanol tolerance, four owned high content of cell wall polysaccharides, and four had a high surface hydrophobicity. Some advantages of these strains as production hosts were revealed. Furthermore, the deletion strains with high surface hydrophobicity were used as hosts to display Candida antarctica lipase B (CALB). The strain gcw22Δ/CALB-GCW61 showed excellent fermentation characteristics, including a faster growth rate and higher hydrolytic activity. CONCLUSIONS: This GPI deletion library has some potential applications for production strains and offers a valuable resource for studying the precise functions of GPI proteins, especially their putative functions. Text Antarc* Antarctica PubMed Central (PMC) BMC Microbiology 20 1 |
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Research Article Wang, Pan Lin, Ying Zou, Chengjuan Zhao, Fengguang Liang, Shuli Zheng, Suiping Han, Shuangyan Construction and screening of a glycosylphosphatidylinositol protein deletion library in Pichia pastoris |
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Research Article |
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BACKGROUND: Glycosylphosphatidylinositol (GPI)-anchored glycoproteins have diverse intrinsic functions in yeasts, and they also have different uses in vitro. In this study, the functions of potential GPI proteins in Pichia pastoris were explored by gene knockout approaches. RESULTS: Through an extensive knockout of GPI proteins in P. pastoris, a single-gene deletion library was constructed for 45 predicted GPI proteins. The knockout of proteins may lead to the activation of a cellular response named the ‘compensatory mechanism’, which is characterized by changes in the content and relationship between cell wall polysaccharides and surface proteins. Among the 45 deletion strains, five showed obvious methanol tolerance, four owned high content of cell wall polysaccharides, and four had a high surface hydrophobicity. Some advantages of these strains as production hosts were revealed. Furthermore, the deletion strains with high surface hydrophobicity were used as hosts to display Candida antarctica lipase B (CALB). The strain gcw22Δ/CALB-GCW61 showed excellent fermentation characteristics, including a faster growth rate and higher hydrolytic activity. CONCLUSIONS: This GPI deletion library has some potential applications for production strains and offers a valuable resource for studying the precise functions of GPI proteins, especially their putative functions. |
format |
Text |
author |
Wang, Pan Lin, Ying Zou, Chengjuan Zhao, Fengguang Liang, Shuli Zheng, Suiping Han, Shuangyan |
author_facet |
Wang, Pan Lin, Ying Zou, Chengjuan Zhao, Fengguang Liang, Shuli Zheng, Suiping Han, Shuangyan |
author_sort |
Wang, Pan |
title |
Construction and screening of a glycosylphosphatidylinositol protein deletion library in Pichia pastoris |
title_short |
Construction and screening of a glycosylphosphatidylinositol protein deletion library in Pichia pastoris |
title_full |
Construction and screening of a glycosylphosphatidylinositol protein deletion library in Pichia pastoris |
title_fullStr |
Construction and screening of a glycosylphosphatidylinositol protein deletion library in Pichia pastoris |
title_full_unstemmed |
Construction and screening of a glycosylphosphatidylinositol protein deletion library in Pichia pastoris |
title_sort |
construction and screening of a glycosylphosphatidylinositol protein deletion library in pichia pastoris |
publisher |
BioMed Central |
publishDate |
2020 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7446130/ http://www.ncbi.nlm.nih.gov/pubmed/32838766 https://doi.org/10.1186/s12866-020-01928-y |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
BMC Microbiol |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7446130/ http://www.ncbi.nlm.nih.gov/pubmed/32838766 http://dx.doi.org/10.1186/s12866-020-01928-y |
op_rights |
© The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. |
op_rightsnorm |
CC0 PDM CC-BY |
op_doi |
https://doi.org/10.1186/s12866-020-01928-y |
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BMC Microbiology |
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20 |
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