Characterization of a Thermophilic and Inhibitor-Tolerant GH1 β-Glucosidase Present in a Hot Spring
β-glucosidase is a key enzyme in the degradation of lignocellulosic biomass, which is responsible for the conversion of oligosaccharides from cellulose hydrolysates to glucose. However, its required high temperatures and the presence of inhibitors have limited its use in industry. In this study, a n...
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MDPI AG
2023-09-01
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| Online Access: | https://www.mdpi.com/2073-4441/15/19/3389 |
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| author | Yu-Ying Huang Pei Wu Xing-Ci Wu Qian-Ru Zhu Qian Zhu Hong-Zhao Zheng Dan Zhu Zhi-Hua Lv Yi-Rui Yin |
| author_facet | Yu-Ying Huang Pei Wu Xing-Ci Wu Qian-Ru Zhu Qian Zhu Hong-Zhao Zheng Dan Zhu Zhi-Hua Lv Yi-Rui Yin |
| author_sort | Yu-Ying Huang |
| collection | DOAJ |
| description | β-glucosidase is a key enzyme in the degradation of lignocellulosic biomass, which is responsible for the conversion of oligosaccharides from cellulose hydrolysates to glucose. However, its required high temperatures and the presence of inhibitors have limited its use in industry. In this study, a new β-glucosidase gene, named <i>thbg2</i>, was obtained from the metagenome Ruidian Hot Spring, Tengchong City, Yunnan Province, southwestern China. The gene was synthesized, cloned, heterologously expressed, and enzymatically characterized. Its optimum temperature and pH were 60 °C and pH 5.6, respectively. ThBg2 exhibited more than 60% relative activity in temperatures ranging from 40 °C to 70 °C and across a pH of 4.0–6.6. It maintained 100% relative activity after incubation at either 50 °C for 24 h or 60 °C for 12 h and more than 80% residual activity after incubation at pH 4.0–6.0 for 24 h. Moreover, it maintained more than 80% relative activity in the presence of heavy metal ions, ethanol, SDS etc. Furthermore, glucose yields from corn stalks increased by 20% after ThBg2 (0.05 mg/mL) was added to the commercial cellulase reaction system. Overall, this work identified a thermophilic and inhibitor-tolerant β-glucosidase with potential applications in commercial lignocellulose utilization and the bioenergy industry. |
| first_indexed | 2024-03-09T13:51:11Z |
| format | Article |
| id | doaj.art-791a310ec27f44c39da362bb83770293 |
| institution | Directory Open Access Journal |
| issn | 2073-4441 |
| language | English |
| last_indexed | 2024-03-09T13:51:11Z |
| publishDate | 2023-09-01 |
| publisher | MDPI AG |
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| series | Water |
| spelling | doaj.art-791a310ec27f44c39da362bb837702932023-11-30T20:49:22ZengMDPI AGWater2073-44412023-09-011519338910.3390/w15193389Characterization of a Thermophilic and Inhibitor-Tolerant GH1 β-Glucosidase Present in a Hot SpringYu-Ying Huang0Pei Wu1Xing-Ci Wu2Qian-Ru Zhu3Qian Zhu4Hong-Zhao Zheng5Dan Zhu6Zhi-Hua Lv7Yi-Rui Yin8College of Agriculture and Biological Science, Dali University, Dali 671003, ChinaCollege of Agriculture and Biological Science, Dali University, Dali 671003, ChinaCollege of Agriculture and Biological Science, Dali University, Dali 671003, ChinaCollege of Agriculture and Biological Science, Dali University, Dali 671003, ChinaCollege of Agriculture and Biological Science, Dali University, Dali 671003, ChinaCollege of Agriculture and Biological Science, Dali University, Dali 671003, ChinaCollege of Agriculture and Biological Science, Dali University, Dali 671003, ChinaCollege of Agriculture and Biological Science, Dali University, Dali 671003, ChinaCollege of Agriculture and Biological Science, Dali University, Dali 671003, Chinaβ-glucosidase is a key enzyme in the degradation of lignocellulosic biomass, which is responsible for the conversion of oligosaccharides from cellulose hydrolysates to glucose. However, its required high temperatures and the presence of inhibitors have limited its use in industry. In this study, a new β-glucosidase gene, named <i>thbg2</i>, was obtained from the metagenome Ruidian Hot Spring, Tengchong City, Yunnan Province, southwestern China. The gene was synthesized, cloned, heterologously expressed, and enzymatically characterized. Its optimum temperature and pH were 60 °C and pH 5.6, respectively. ThBg2 exhibited more than 60% relative activity in temperatures ranging from 40 °C to 70 °C and across a pH of 4.0–6.6. It maintained 100% relative activity after incubation at either 50 °C for 24 h or 60 °C for 12 h and more than 80% residual activity after incubation at pH 4.0–6.0 for 24 h. Moreover, it maintained more than 80% relative activity in the presence of heavy metal ions, ethanol, SDS etc. Furthermore, glucose yields from corn stalks increased by 20% after ThBg2 (0.05 mg/mL) was added to the commercial cellulase reaction system. Overall, this work identified a thermophilic and inhibitor-tolerant β-glucosidase with potential applications in commercial lignocellulose utilization and the bioenergy industry.https://www.mdpi.com/2073-4441/15/19/3389hot springmetagenomethermophilicheavy metal ion tolerantglucose tolerantβ-glucosidase |
| spellingShingle | Yu-Ying Huang Pei Wu Xing-Ci Wu Qian-Ru Zhu Qian Zhu Hong-Zhao Zheng Dan Zhu Zhi-Hua Lv Yi-Rui Yin Characterization of a Thermophilic and Inhibitor-Tolerant GH1 β-Glucosidase Present in a Hot Spring Water hot spring metagenome thermophilic heavy metal ion tolerant glucose tolerant β-glucosidase |
| title | Characterization of a Thermophilic and Inhibitor-Tolerant GH1 β-Glucosidase Present in a Hot Spring |
| title_full | Characterization of a Thermophilic and Inhibitor-Tolerant GH1 β-Glucosidase Present in a Hot Spring |
| title_fullStr | Characterization of a Thermophilic and Inhibitor-Tolerant GH1 β-Glucosidase Present in a Hot Spring |
| title_full_unstemmed | Characterization of a Thermophilic and Inhibitor-Tolerant GH1 β-Glucosidase Present in a Hot Spring |
| title_short | Characterization of a Thermophilic and Inhibitor-Tolerant GH1 β-Glucosidase Present in a Hot Spring |
| title_sort | characterization of a thermophilic and inhibitor tolerant gh1 β glucosidase present in a hot spring |
| topic | hot spring metagenome thermophilic heavy metal ion tolerant glucose tolerant β-glucosidase |
| url | https://www.mdpi.com/2073-4441/15/19/3389 |
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