Engineering Cell Polarization Improves Protein Production in <i>Saccharomyces cerevisiae</i>
<i>Saccharomyces cerevisiae</i> has been widely used as a microbial cell factory to produce recombinant proteins. Therefore, enhancing the protein production efficiency of yeast cell factories to expand the market demand for protein products is necessary. Recombinant proteins are often r...
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MDPI AG
2022-10-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/10/10/2005 |
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| author | Shuo Yang Junfeng Shen Jiliang Deng Hongxing Li Jianzhi Zhao Hongting Tang Xiaoming Bao |
| author_facet | Shuo Yang Junfeng Shen Jiliang Deng Hongxing Li Jianzhi Zhao Hongting Tang Xiaoming Bao |
| author_sort | Shuo Yang |
| collection | DOAJ |
| description | <i>Saccharomyces cerevisiae</i> has been widely used as a microbial cell factory to produce recombinant proteins. Therefore, enhancing the protein production efficiency of yeast cell factories to expand the market demand for protein products is necessary. Recombinant proteins are often retained in the secretory pathway because of the limited protein transport performed by vesicle trafficking. Cell polarization describes the asymmetric organization of the plasma membrane cytoskeleton and organelles and tightly regulates vesicle trafficking for protein transport. Engineering vesicle trafficking has broadly been studied by the overexpression or deletion of key genes involved but not by modifying cell polarization. Here, we used α-amylase as a reporter protein, and its secretion and surface-display were first improved by promoter optimization. To study the effect of engineering cell polarization on protein production, fourteen genes related to cell polarization were overexpressed. <i>BUD1</i>, <i>CDC42</i>, <i>AXL1</i>, and <i>BUD10</i> overexpression increased the activity of surface-displayed α-amylase, and <i>BUD1</i>, <i>BUD3</i>, <i>BUD4</i>, <i>BUD7</i>, and <i>BUD10</i> overexpression enhanced secreted α-amylase activity. Furthermore, <i>BUD1</i> overexpression increased the surface-displayed and secreted α-amylase expression by 56% and 49%, respectively. We also observed that the combinatorial modification and regulation of gene expression improved α-amylase production in a dose-dependent manner. <i>BUD1</i> and <i>CDC42</i> co-overexpression increased the α-amylase surface display by 100%, and two genomic copies of <i>BUD1</i> improved α-amylase secretion by 92%. Furthermore, these modifications were used to improve the surface display and secretion of the recombinant β-glucosidase protein. Our study affords a novel insight for improving the surface display and secretion of recombinant proteins. |
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| language | English |
| last_indexed | 2024-03-09T19:44:42Z |
| publishDate | 2022-10-01 |
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| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-ba02257db4b14dbb93b83d6314993ac12023-11-24T01:26:52ZengMDPI AGMicroorganisms2076-26072022-10-011010200510.3390/microorganisms10102005Engineering Cell Polarization Improves Protein Production in <i>Saccharomyces cerevisiae</i>Shuo Yang0Junfeng Shen1Jiliang Deng2Hongxing Li3Jianzhi Zhao4Hongting Tang5Xiaoming Bao6State Key Laboratory of Biobased Material and Green Papermaking, School of Bioengineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250353, ChinaCenter for Synthetic Biochemistry, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes for Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, ChinaCenter for Synthetic Biochemistry, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes for Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, ChinaState Key Laboratory of Biobased Material and Green Papermaking, School of Bioengineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250353, ChinaState Key Laboratory of Biobased Material and Green Papermaking, School of Bioengineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250353, ChinaCenter for Synthetic Biochemistry, Shenzhen Institute of Synthetic Biology, Shenzhen Institutes for Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, ChinaState Key Laboratory of Biobased Material and Green Papermaking, School of Bioengineering, Qilu University of Technology (Shandong Academy of Sciences), Jinan 250353, China<i>Saccharomyces cerevisiae</i> has been widely used as a microbial cell factory to produce recombinant proteins. Therefore, enhancing the protein production efficiency of yeast cell factories to expand the market demand for protein products is necessary. Recombinant proteins are often retained in the secretory pathway because of the limited protein transport performed by vesicle trafficking. Cell polarization describes the asymmetric organization of the plasma membrane cytoskeleton and organelles and tightly regulates vesicle trafficking for protein transport. Engineering vesicle trafficking has broadly been studied by the overexpression or deletion of key genes involved but not by modifying cell polarization. Here, we used α-amylase as a reporter protein, and its secretion and surface-display were first improved by promoter optimization. To study the effect of engineering cell polarization on protein production, fourteen genes related to cell polarization were overexpressed. <i>BUD1</i>, <i>CDC42</i>, <i>AXL1</i>, and <i>BUD10</i> overexpression increased the activity of surface-displayed α-amylase, and <i>BUD1</i>, <i>BUD3</i>, <i>BUD4</i>, <i>BUD7</i>, and <i>BUD10</i> overexpression enhanced secreted α-amylase activity. Furthermore, <i>BUD1</i> overexpression increased the surface-displayed and secreted α-amylase expression by 56% and 49%, respectively. We also observed that the combinatorial modification and regulation of gene expression improved α-amylase production in a dose-dependent manner. <i>BUD1</i> and <i>CDC42</i> co-overexpression increased the α-amylase surface display by 100%, and two genomic copies of <i>BUD1</i> improved α-amylase secretion by 92%. Furthermore, these modifications were used to improve the surface display and secretion of the recombinant β-glucosidase protein. Our study affords a novel insight for improving the surface display and secretion of recombinant proteins.https://www.mdpi.com/2076-2607/10/10/2005<i>Saccharomyces cerevisiae</i>recombinant proteinscell polarizationsurface-displaysecretion |
| spellingShingle | Shuo Yang Junfeng Shen Jiliang Deng Hongxing Li Jianzhi Zhao Hongting Tang Xiaoming Bao Engineering Cell Polarization Improves Protein Production in <i>Saccharomyces cerevisiae</i> Microorganisms <i>Saccharomyces cerevisiae</i> recombinant proteins cell polarization surface-display secretion |
| title | Engineering Cell Polarization Improves Protein Production in <i>Saccharomyces cerevisiae</i> |
| title_full | Engineering Cell Polarization Improves Protein Production in <i>Saccharomyces cerevisiae</i> |
| title_fullStr | Engineering Cell Polarization Improves Protein Production in <i>Saccharomyces cerevisiae</i> |
| title_full_unstemmed | Engineering Cell Polarization Improves Protein Production in <i>Saccharomyces cerevisiae</i> |
| title_short | Engineering Cell Polarization Improves Protein Production in <i>Saccharomyces cerevisiae</i> |
| title_sort | engineering cell polarization improves protein production in i saccharomyces cerevisiae i |
| topic | <i>Saccharomyces cerevisiae</i> recombinant proteins cell polarization surface-display secretion |
| url | https://www.mdpi.com/2076-2607/10/10/2005 |
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