Biochemical Characterization of Novel GH6 Endoglucanase from <i>Myxococcus</i> sp. B6-1 and Its Effects on Agricultural Straws Saccharification
Cellulase has been widely used in many industrial fields, such as feed and food industry, because it can hydrolyze cellulose to oligosaccharides with a lower degree of polymerization. Endo-β-1,4-glucanase is a critical speed-limiting cellulase in the saccharification process. In this study, endo-β-1...
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MDPI AG
2023-06-01
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| author | Zhen Huang Guorong Ni Longhua Dai Weiqi Zhang Siting Feng Fei Wang |
| author_facet | Zhen Huang Guorong Ni Longhua Dai Weiqi Zhang Siting Feng Fei Wang |
| author_sort | Zhen Huang |
| collection | DOAJ |
| description | Cellulase has been widely used in many industrial fields, such as feed and food industry, because it can hydrolyze cellulose to oligosaccharides with a lower degree of polymerization. Endo-β-1,4-glucanase is a critical speed-limiting cellulase in the saccharification process. In this study, endo-β-1,4-glucanase gene (<i>CelA257</i>) from <i>Myxococcus</i> sp. B6-1 was cloned and expressed in <i>Escherichia coli</i>. CelA257 contained carbohydrate-binding module (CBM) 4-9 and glycosyl hydrolase (GH) family 6 domain that shares 54.7% identity with endoglucanase from <i>Streptomyces halstedii</i>. The recombinant enzyme exhibited optimal activity at pH 6.5 and 50 °C and was stable over a broad pH (6–9.5) range and temperature < 50 °C. CelA257 exhibited broad substrate specificity to barley β-glucan, lichenin, CMC, chitosan, laminarin, avicel, and phosphoric acid swollen cellulose (PASC). CelA257 degraded both cellotetrose (G<sub>4</sub>) and cellppentaose (G<sub>5</sub>) to cellobiose (G<sub>2</sub>) and cellotriose (G<sub>3</sub>). Adding CelA257 increased the release of reducing sugars in crop straw powers, including wheat straw (0.18 mg/mL), rape straw (0.42 mg/mL), rice straw (0.16 mg/mL), peanut straw (0.16 mg/mL), and corn straw (0.61 mg/mL). This study provides a potential additive in biomass saccharification applications. |
| first_indexed | 2024-03-11T01:41:02Z |
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| last_indexed | 2024-03-11T01:41:02Z |
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| spelling | doaj.art-646e2204e87d412181da2dedcb15170c2023-11-18T16:33:40ZengMDPI AGFoods2304-81582023-06-011213251710.3390/foods12132517Biochemical Characterization of Novel GH6 Endoglucanase from <i>Myxococcus</i> sp. B6-1 and Its Effects on Agricultural Straws SaccharificationZhen Huang0Guorong Ni1Longhua Dai2Weiqi Zhang3Siting Feng4Fei Wang5College of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang 330045, ChinaCollege of Land Resources and Environment, Jiangxi Agriculture University, Nanchang 330045, ChinaCollege of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang 330045, ChinaCollege of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang 330045, ChinaCollege of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang 330045, ChinaCollege of Bioscience and Bioengineering, Jiangxi Agricultural University, Nanchang 330045, ChinaCellulase has been widely used in many industrial fields, such as feed and food industry, because it can hydrolyze cellulose to oligosaccharides with a lower degree of polymerization. Endo-β-1,4-glucanase is a critical speed-limiting cellulase in the saccharification process. In this study, endo-β-1,4-glucanase gene (<i>CelA257</i>) from <i>Myxococcus</i> sp. B6-1 was cloned and expressed in <i>Escherichia coli</i>. CelA257 contained carbohydrate-binding module (CBM) 4-9 and glycosyl hydrolase (GH) family 6 domain that shares 54.7% identity with endoglucanase from <i>Streptomyces halstedii</i>. The recombinant enzyme exhibited optimal activity at pH 6.5 and 50 °C and was stable over a broad pH (6–9.5) range and temperature < 50 °C. CelA257 exhibited broad substrate specificity to barley β-glucan, lichenin, CMC, chitosan, laminarin, avicel, and phosphoric acid swollen cellulose (PASC). CelA257 degraded both cellotetrose (G<sub>4</sub>) and cellppentaose (G<sub>5</sub>) to cellobiose (G<sub>2</sub>) and cellotriose (G<sub>3</sub>). Adding CelA257 increased the release of reducing sugars in crop straw powers, including wheat straw (0.18 mg/mL), rape straw (0.42 mg/mL), rice straw (0.16 mg/mL), peanut straw (0.16 mg/mL), and corn straw (0.61 mg/mL). This study provides a potential additive in biomass saccharification applications.https://www.mdpi.com/2304-8158/12/13/2517endo-β-1,4-glucanase<i>Escherichia coli</i>expressioncharacterizationoligosaccharidesaccharification |
| spellingShingle | Zhen Huang Guorong Ni Longhua Dai Weiqi Zhang Siting Feng Fei Wang Biochemical Characterization of Novel GH6 Endoglucanase from <i>Myxococcus</i> sp. B6-1 and Its Effects on Agricultural Straws Saccharification Foods endo-β-1,4-glucanase <i>Escherichia coli</i> expression characterization oligosaccharide saccharification |
| title | Biochemical Characterization of Novel GH6 Endoglucanase from <i>Myxococcus</i> sp. B6-1 and Its Effects on Agricultural Straws Saccharification |
| title_full | Biochemical Characterization of Novel GH6 Endoglucanase from <i>Myxococcus</i> sp. B6-1 and Its Effects on Agricultural Straws Saccharification |
| title_fullStr | Biochemical Characterization of Novel GH6 Endoglucanase from <i>Myxococcus</i> sp. B6-1 and Its Effects on Agricultural Straws Saccharification |
| title_full_unstemmed | Biochemical Characterization of Novel GH6 Endoglucanase from <i>Myxococcus</i> sp. B6-1 and Its Effects on Agricultural Straws Saccharification |
| title_short | Biochemical Characterization of Novel GH6 Endoglucanase from <i>Myxococcus</i> sp. B6-1 and Its Effects on Agricultural Straws Saccharification |
| title_sort | biochemical characterization of novel gh6 endoglucanase from i myxococcus i sp b6 1 and its effects on agricultural straws saccharification |
| topic | endo-β-1,4-glucanase <i>Escherichia coli</i> expression characterization oligosaccharide saccharification |
| url | https://www.mdpi.com/2304-8158/12/13/2517 |
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