Overproduction of native endo-β-1,4-glucanases leads to largely enhanced biomass saccharification and bioethanol production by specific modification of cellulose features in transgenic rice
Abstract Background Genetic modification of plant cell walls has been implemented to reduce lignocellulosic recalcitrance for biofuel production. Plant glycoside hydrolase family 9 (GH9) comprises endo-β-1,4-glucanase in plants. Few studies have examined the roles of GH9 in cell wall modification. I...
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| Format: | Article |
| Language: | English |
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BMC
2019-01-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13068-018-1351-1 |
| _version_ | 1818218797940604928 |
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| author | Jiangfeng Huang Tao Xia Guanhua Li Xianliang Li Ying Li Yanting Wang Youmei Wang Yuanyuan Chen Guosheng Xie Feng-Wu Bai Liangcai Peng Lingqiang Wang |
| author_facet | Jiangfeng Huang Tao Xia Guanhua Li Xianliang Li Ying Li Yanting Wang Youmei Wang Yuanyuan Chen Guosheng Xie Feng-Wu Bai Liangcai Peng Lingqiang Wang |
| author_sort | Jiangfeng Huang |
| collection | DOAJ |
| description | Abstract Background Genetic modification of plant cell walls has been implemented to reduce lignocellulosic recalcitrance for biofuel production. Plant glycoside hydrolase family 9 (GH9) comprises endo-β-1,4-glucanase in plants. Few studies have examined the roles of GH9 in cell wall modification. In this study, we independently overexpressed two genes from GH9B subclasses (OsGH9B1 and OsGH9B3) and examined cell wall features and biomass saccharification in transgenic rice plants. Results Compared with the wild type (WT, Nipponbare), the OsGH9B1 and OsGH9B3 transgenic rice plants, respectively, contained much higher OsGH9B1 and OsGH9B3 protein levels and both proteins were observed in situ with nonspecific distribution in the plant cells. The transgenic lines exhibited significantly increased cellulase activity in vitro than the WT. The OsGH9B1 and OsGH9B3 transgenic plants showed a slight alteration in three wall polymer compositions (cellulose, hemicelluloses, and lignin), in their stem mechanical strength and biomass yield, but were significantly decreased in the cellulose degree of polymerization (DP) and lignocellulose crystalline index (CrI) by 21–22%. Notably, the crude cellulose substrates of the transgenic lines were more efficiently digested by cellobiohydrolase (CBHI) than those of the WT, indicating the significantly increased amounts of reducing ends of β-1,4-glucans in cellulose microfibrils. Finally, the engineered lines generated high sugar yields after mild alkali pretreatments and subsequent enzymatic hydrolysis, resulting in the high bioethanol yields obtained at 22.5% of dry matter. Conclusions Overproduction of OsGH9B1/B3 enzymes should have specific activity in the postmodification of cellulose microfibrils. The increased reducing ends of β-1,4-glucan chains for reduced cellulose DP and CrI positively affected biomass enzymatic saccharification. Our results demonstrate a potential strategy for genetic modification of cellulose microfibrils in bioenergy crops. |
| first_indexed | 2024-12-12T07:29:29Z |
| format | Article |
| id | doaj.art-994c5648482a44af9f2efcd5c1250047 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-12-12T07:29:29Z |
| publishDate | 2019-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-994c5648482a44af9f2efcd5c12500472022-12-22T00:33:04ZengBMCBiotechnology for Biofuels1754-68342019-01-0112111510.1186/s13068-018-1351-1Overproduction of native endo-β-1,4-glucanases leads to largely enhanced biomass saccharification and bioethanol production by specific modification of cellulose features in transgenic riceJiangfeng Huang0Tao Xia1Guanhua Li2Xianliang Li3Ying Li4Yanting Wang5Youmei Wang6Yuanyuan Chen7Guosheng Xie8Feng-Wu Bai9Liangcai Peng10Lingqiang Wang11Biomass and Bioenergy Research Centre, College of Plant Science and Technology, College of Life Science and Technology, Huazhong Agricultural UniversityBiomass and Bioenergy Research Centre, College of Plant Science and Technology, College of Life Science and Technology, Huazhong Agricultural UniversityState Key Laboratory of Reproductive Regulation and Breeding of Grassland Livestock, School of Life Sciences, Inner Mongolia UniversityCollege of Bioengineering, Jingchu University of TechnologyBiomass and Bioenergy Research Centre, College of Plant Science and Technology, College of Life Science and Technology, Huazhong Agricultural UniversityBiomass and Bioenergy Research Centre, College of Plant Science and Technology, College of Life Science and Technology, Huazhong Agricultural UniversityBiomass and Bioenergy Research Centre, College of Plant Science and Technology, College of Life Science and Technology, Huazhong Agricultural UniversityBiomass and Bioenergy Research Centre, College of Plant Science and Technology, College of Life Science and Technology, Huazhong Agricultural UniversityBiomass and Bioenergy Research Centre, College of Plant Science and Technology, College of Life Science and Technology, Huazhong Agricultural UniversityState Key Laboratory of Microbial Metabolism, School of Life Science and Biotechnology, Shanghai Jiao Tong UniversityBiomass and Bioenergy Research Centre, College of Plant Science and Technology, College of Life Science and Technology, Huazhong Agricultural UniversityBiomass and Bioenergy Research Centre, College of Plant Science and Technology, College of Life Science and Technology, Huazhong Agricultural UniversityAbstract Background Genetic modification of plant cell walls has been implemented to reduce lignocellulosic recalcitrance for biofuel production. Plant glycoside hydrolase family 9 (GH9) comprises endo-β-1,4-glucanase in plants. Few studies have examined the roles of GH9 in cell wall modification. In this study, we independently overexpressed two genes from GH9B subclasses (OsGH9B1 and OsGH9B3) and examined cell wall features and biomass saccharification in transgenic rice plants. Results Compared with the wild type (WT, Nipponbare), the OsGH9B1 and OsGH9B3 transgenic rice plants, respectively, contained much higher OsGH9B1 and OsGH9B3 protein levels and both proteins were observed in situ with nonspecific distribution in the plant cells. The transgenic lines exhibited significantly increased cellulase activity in vitro than the WT. The OsGH9B1 and OsGH9B3 transgenic plants showed a slight alteration in three wall polymer compositions (cellulose, hemicelluloses, and lignin), in their stem mechanical strength and biomass yield, but were significantly decreased in the cellulose degree of polymerization (DP) and lignocellulose crystalline index (CrI) by 21–22%. Notably, the crude cellulose substrates of the transgenic lines were more efficiently digested by cellobiohydrolase (CBHI) than those of the WT, indicating the significantly increased amounts of reducing ends of β-1,4-glucans in cellulose microfibrils. Finally, the engineered lines generated high sugar yields after mild alkali pretreatments and subsequent enzymatic hydrolysis, resulting in the high bioethanol yields obtained at 22.5% of dry matter. Conclusions Overproduction of OsGH9B1/B3 enzymes should have specific activity in the postmodification of cellulose microfibrils. The increased reducing ends of β-1,4-glucan chains for reduced cellulose DP and CrI positively affected biomass enzymatic saccharification. Our results demonstrate a potential strategy for genetic modification of cellulose microfibrils in bioenergy crops.http://link.springer.com/article/10.1186/s13068-018-1351-1Endo-β-1,4-glucanasesTransgenic riceGH9BCellulose modificationBiomass saccharificationBioethanol production |
| spellingShingle | Jiangfeng Huang Tao Xia Guanhua Li Xianliang Li Ying Li Yanting Wang Youmei Wang Yuanyuan Chen Guosheng Xie Feng-Wu Bai Liangcai Peng Lingqiang Wang Overproduction of native endo-β-1,4-glucanases leads to largely enhanced biomass saccharification and bioethanol production by specific modification of cellulose features in transgenic rice Biotechnology for Biofuels Endo-β-1,4-glucanases Transgenic rice GH9B Cellulose modification Biomass saccharification Bioethanol production |
| title | Overproduction of native endo-β-1,4-glucanases leads to largely enhanced biomass saccharification and bioethanol production by specific modification of cellulose features in transgenic rice |
| title_full | Overproduction of native endo-β-1,4-glucanases leads to largely enhanced biomass saccharification and bioethanol production by specific modification of cellulose features in transgenic rice |
| title_fullStr | Overproduction of native endo-β-1,4-glucanases leads to largely enhanced biomass saccharification and bioethanol production by specific modification of cellulose features in transgenic rice |
| title_full_unstemmed | Overproduction of native endo-β-1,4-glucanases leads to largely enhanced biomass saccharification and bioethanol production by specific modification of cellulose features in transgenic rice |
| title_short | Overproduction of native endo-β-1,4-glucanases leads to largely enhanced biomass saccharification and bioethanol production by specific modification of cellulose features in transgenic rice |
| title_sort | overproduction of native endo β 1 4 glucanases leads to largely enhanced biomass saccharification and bioethanol production by specific modification of cellulose features in transgenic rice |
| topic | Endo-β-1,4-glucanases Transgenic rice GH9B Cellulose modification Biomass saccharification Bioethanol production |
| url | http://link.springer.com/article/10.1186/s13068-018-1351-1 |
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