Construction and screening of a glycosylphosphatidylinositol protein deletion library in Pichia pastoris
Abstract 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 a...
Published in: | BMC Microbiology |
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crspringernat:10.1186/s12866-020-01928-y 2023-05-15T14:11:58+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 National Natural Science Foundation of China 2020 http://dx.doi.org/10.1186/s12866-020-01928-y https://link.springer.com/content/pdf/10.1186/s12866-020-01928-y.pdf https://link.springer.com/article/10.1186/s12866-020-01928-y/fulltext.html en eng Springer Science and Business Media LLC https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 CC-BY BMC Microbiology volume 20, issue 1 ISSN 1471-2180 Microbiology (medical) Microbiology journal-article 2020 crspringernat https://doi.org/10.1186/s12866-020-01928-y 2022-01-04T14:20:48Z Abstract 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. Article in Journal/Newspaper Antarc* Antarctica Springer Nature (via Crossref) BMC Microbiology 20 1 |
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Springer Nature (via Crossref) |
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crspringernat |
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English |
topic |
Microbiology (medical) Microbiology |
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Microbiology (medical) Microbiology 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 |
topic_facet |
Microbiology (medical) Microbiology |
description |
Abstract 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. |
author2 |
National Natural Science Foundation of China |
format |
Article in Journal/Newspaper |
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 |
Springer Science and Business Media LLC |
publishDate |
2020 |
url |
http://dx.doi.org/10.1186/s12866-020-01928-y https://link.springer.com/content/pdf/10.1186/s12866-020-01928-y.pdf https://link.springer.com/article/10.1186/s12866-020-01928-y/fulltext.html |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
BMC Microbiology volume 20, issue 1 ISSN 1471-2180 |
op_rights |
https://creativecommons.org/licenses/by/4.0 https://creativecommons.org/licenses/by/4.0 |
op_rightsnorm |
CC-BY |
op_doi |
https://doi.org/10.1186/s12866-020-01928-y |
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BMC Microbiology |
container_volume |
20 |
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1 |
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1766284207786557440 |