Overexpression of LAS21 in Cellulase-Displaying <i>Saccharomyces cerevisiae</i> for High-Yield Ethanol Production from Pretreated Sugarcane Bagasse
The valorization of lignocellulosic feedstocks into biofuels and biochemicals has received much attention due to its environmental friendliness and sustainability. However, engineering an ideal microorganism that can both produce sufficient cellulases and ferment ethanol is highly challenging. In th...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-11-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/8/11/652 |
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| author | Jantima Arnthong Piyada Bussadee Apisan Phienluphon Pacharawan Deenarn Kan Tulsook Sa-ngapong Plupjeen Chatuphon Siamphan Chakrit Tachaapaikoon Verawat Champreda Surisa Suwannarangsee |
| author_facet | Jantima Arnthong Piyada Bussadee Apisan Phienluphon Pacharawan Deenarn Kan Tulsook Sa-ngapong Plupjeen Chatuphon Siamphan Chakrit Tachaapaikoon Verawat Champreda Surisa Suwannarangsee |
| author_sort | Jantima Arnthong |
| collection | DOAJ |
| description | The valorization of lignocellulosic feedstocks into biofuels and biochemicals has received much attention due to its environmental friendliness and sustainability. However, engineering an ideal microorganism that can both produce sufficient cellulases and ferment ethanol is highly challenging. In this study, we have tested seven different genes that are involved in glycosylphosphatidylinositol (GPI) biosynthesis and remodeling for the improvement of cellulase activity tethered on the <i>S. cerevisiae</i> cell surface. It was found that the overexpression of LAS21 can improve β-glucosidase activity by 48.8% compared to the original strain. Then, the three cellulase genes (cellobiohydrolase, endoglucanase, and β-glucosidase) and the LAS21 gene were co-introduced into a diploid thermotolerant <i>S. cerevisiae</i> strain by a multiple-round transformation approach, resulting in the cellulolytic ECBLCCE5 strain. Further optimization of the bioprocess parameters was found to enhance the ethanol yield of the ECBLCCE5 strain. Scaling up the valorization of pretreated sugarcane bagasses in a 1 L bioreactor resulted in a maximum ethanol concentration of 28.0 g/L (86.5% of theoretical yield). Our study provides a promising way to improve the economic viability of second-generation ethanol production. Moreover, the engineering of genes involved in GPI biosynthesis and remodeling can be applied to other yeast cell surface display applications. |
| first_indexed | 2024-03-09T18:20:39Z |
| format | Article |
| id | doaj.art-752eb440227e434ab813be7c5828708e |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-09T18:20:39Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-752eb440227e434ab813be7c5828708e2023-11-24T08:19:57ZengMDPI AGFermentation2311-56372022-11-0181165210.3390/fermentation8110652Overexpression of LAS21 in Cellulase-Displaying <i>Saccharomyces cerevisiae</i> for High-Yield Ethanol Production from Pretreated Sugarcane BagasseJantima Arnthong0Piyada Bussadee1Apisan Phienluphon2Pacharawan Deenarn3Kan Tulsook4Sa-ngapong Plupjeen5Chatuphon Siamphan6Chakrit Tachaapaikoon7Verawat Champreda8Surisa Suwannarangsee9National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Klong Luang, Pathumthani 12120, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Klong Luang, Pathumthani 12120, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Klong Luang, Pathumthani 12120, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Klong Luang, Pathumthani 12120, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Klong Luang, Pathumthani 12120, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Klong Luang, Pathumthani 12120, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Klong Luang, Pathumthani 12120, ThailandExcellent Center of Enzyme Technology and Microbial Utilization, Pilot Plant Development and Training Institute, King Mongkut’s University of Technology Thonburi, Bangkok 10150, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Klong Luang, Pathumthani 12120, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), 113 Thailand Science Park, Klong Luang, Pathumthani 12120, ThailandThe valorization of lignocellulosic feedstocks into biofuels and biochemicals has received much attention due to its environmental friendliness and sustainability. However, engineering an ideal microorganism that can both produce sufficient cellulases and ferment ethanol is highly challenging. In this study, we have tested seven different genes that are involved in glycosylphosphatidylinositol (GPI) biosynthesis and remodeling for the improvement of cellulase activity tethered on the <i>S. cerevisiae</i> cell surface. It was found that the overexpression of LAS21 can improve β-glucosidase activity by 48.8% compared to the original strain. Then, the three cellulase genes (cellobiohydrolase, endoglucanase, and β-glucosidase) and the LAS21 gene were co-introduced into a diploid thermotolerant <i>S. cerevisiae</i> strain by a multiple-round transformation approach, resulting in the cellulolytic ECBLCCE5 strain. Further optimization of the bioprocess parameters was found to enhance the ethanol yield of the ECBLCCE5 strain. Scaling up the valorization of pretreated sugarcane bagasses in a 1 L bioreactor resulted in a maximum ethanol concentration of 28.0 g/L (86.5% of theoretical yield). Our study provides a promising way to improve the economic viability of second-generation ethanol production. Moreover, the engineering of genes involved in GPI biosynthesis and remodeling can be applied to other yeast cell surface display applications.https://www.mdpi.com/2311-5637/8/11/652<i>Saccharomyces cerevisiae</i>cell surface displaycellulaseLAS21ethanol productionlignocellulosic biomass |
| spellingShingle | Jantima Arnthong Piyada Bussadee Apisan Phienluphon Pacharawan Deenarn Kan Tulsook Sa-ngapong Plupjeen Chatuphon Siamphan Chakrit Tachaapaikoon Verawat Champreda Surisa Suwannarangsee Overexpression of LAS21 in Cellulase-Displaying <i>Saccharomyces cerevisiae</i> for High-Yield Ethanol Production from Pretreated Sugarcane Bagasse Fermentation <i>Saccharomyces cerevisiae</i> cell surface display cellulase LAS21 ethanol production lignocellulosic biomass |
| title | Overexpression of LAS21 in Cellulase-Displaying <i>Saccharomyces cerevisiae</i> for High-Yield Ethanol Production from Pretreated Sugarcane Bagasse |
| title_full | Overexpression of LAS21 in Cellulase-Displaying <i>Saccharomyces cerevisiae</i> for High-Yield Ethanol Production from Pretreated Sugarcane Bagasse |
| title_fullStr | Overexpression of LAS21 in Cellulase-Displaying <i>Saccharomyces cerevisiae</i> for High-Yield Ethanol Production from Pretreated Sugarcane Bagasse |
| title_full_unstemmed | Overexpression of LAS21 in Cellulase-Displaying <i>Saccharomyces cerevisiae</i> for High-Yield Ethanol Production from Pretreated Sugarcane Bagasse |
| title_short | Overexpression of LAS21 in Cellulase-Displaying <i>Saccharomyces cerevisiae</i> for High-Yield Ethanol Production from Pretreated Sugarcane Bagasse |
| title_sort | overexpression of las21 in cellulase displaying i saccharomyces cerevisiae i for high yield ethanol production from pretreated sugarcane bagasse |
| topic | <i>Saccharomyces cerevisiae</i> cell surface display cellulase LAS21 ethanol production lignocellulosic biomass |
| url | https://www.mdpi.com/2311-5637/8/11/652 |
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