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Biochemical characterization of a novel thermo‐halo‐tolerant GH5 endoglucanase from a thermal spring metagenome
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2021-01-06 , DOI: 10.1002/bit.27668 Namrata Joshi 1 , Girija Kaushal 1 , Sudhir P Singh 1
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2021-01-06 , DOI: 10.1002/bit.27668 Namrata Joshi 1 , Girija Kaushal 1 , Sudhir P Singh 1
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A novel endoglucanase gene, celM, was cloned from a thermal spring metagenome. The gene was expressed in Escherichia coli, and the protein was extracted and purified. The protein catalyzed the hydrolysis of amorphous cellulose in a wide range of temperatures, 30–95°C, with optimal activity at 80°C. It was able to tolerate high temperature (80°C) with a half‐life of 8 h. Its activity was eminent in a wide pH range of 3.0–11.0, with the highest activity at pH 6.0. The enzyme was tested for halostability. Any significant loss was not recorded in the activity of CelM after the exposure to salinity (3 M NaCl) for 30 days. Furthermore, CelM displayed a substantial resistance toward metal ions, denaturant, reducing agent, organic solvent, and non‐ionic surfactants. The amorphous cellulose, treated with CelM, was randomly cleaved, generating cello‐oligosaccharides of 2–5 degree of polymerization. Furthermore, CelM was demonstrated to catalyze the hydrolysis of cellulose fraction in the delignified biomass samples, for example, sweet sorghum bagasse, rice straw, and corncob, into cello‐oligosaccharides. Given that CelM is a thermo‐halo‐tolerant GH5 endoglucanase, with resistance to detergents and organic solvent, the biocatalyst could be of potential usefulness for a variety of industrial applications.
中文翻译：
来自温泉宏基因组的新型耐热耐卤 GH5 内切葡聚糖酶的生化表征
从温泉宏基因组中克隆了一种新的内切葡聚糖酶基因cel M。该基因在大肠杆菌中表达，蛋白被提取纯化。该蛋白质在 30-95°C 的宽温度范围内催化无定形纤维素的水解，在 80°C 时具有最佳活性。它能够耐受高温（80°C），半衰期为 8 小时。它的活性在 3.0-11.0 的宽 pH 范围内突出，在 pH 6.0 时活性最高。测试酶的可卤性。在暴露于盐度 (3 M NaCl) 30 天后，Cel M的活性没有记录到任何显着损失。此外，Cel M对金属离子、变性剂、还原剂、有机溶剂和非离子表面活性剂表现出显着的抵抗力。用 Cel M处理的无定形纤维素随机裂解，产生 2-5 聚合度的纤维寡糖。此外，Cel M被证明可催化脱木质生物质样品（例如甜高粱甘蔗渣、稻草和玉米芯）中的纤维素部分水解成纤维寡糖。鉴于 Cel M是一种耐热卤素 GH5 内切葡聚糖酶，对洗涤剂和有机溶剂具有耐受性，因此该生物催化剂可能对各种工业应用具有潜在用途。
更新日期：2021-03-17
中文翻译：
来自温泉宏基因组的新型耐热耐卤 GH5 内切葡聚糖酶的生化表征
从温泉宏基因组中克隆了一种新的内切葡聚糖酶基因cel M。该基因在大肠杆菌中表达，蛋白被提取纯化。该蛋白质在 30-95°C 的宽温度范围内催化无定形纤维素的水解，在 80°C 时具有最佳活性。它能够耐受高温（80°C），半衰期为 8 小时。它的活性在 3.0-11.0 的宽 pH 范围内突出，在 pH 6.0 时活性最高。测试酶的可卤性。在暴露于盐度 (3 M NaCl) 30 天后，Cel M的活性没有记录到任何显着损失。此外，Cel M对金属离子、变性剂、还原剂、有机溶剂和非离子表面活性剂表现出显着的抵抗力。用 Cel M处理的无定形纤维素随机裂解，产生 2-5 聚合度的纤维寡糖。此外，Cel M被证明可催化脱木质生物质样品（例如甜高粱甘蔗渣、稻草和玉米芯）中的纤维素部分水解成纤维寡糖。鉴于 Cel M是一种耐热卤素 GH5 内切葡聚糖酶，对洗涤剂和有机溶剂具有耐受性，因此该生物催化剂可能对各种工业应用具有潜在用途。
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