Expression and characterization of mesophilic GH1 β-glucosidase CdBglA from acidophilic <i>Cuniculiplasma divulgatum</i>.

Jinjian HE; Fengfei Shen; Xinhan Liu; Tianjun Yang; Baotong LI; Pengjun Shi; Huiqin Liu; Wanning Zeng

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Expression and characterization of mesophilic GH1 β-glucosidase CdBglA from acidophilic Cuniculiplasma divulgatum.

Author: Jinjian HE ¹ ; Fengfei SHEN ² ; Xinhan LIU ¹ ; Tianjun YANG ³ ; Baotong LI ¹ ; Pengjun SHI ⁴ ; Huiqin LIU ¹ ; Wanning ZENG ¹
Author Information

1. College of Horticulture and Landscape, Tianjin Agricultural University, Tianjin 300392, China.
2. Jiangxi Zhonghong Boyuan Biotechnology Co., Ltd., Nanchang 330200, Jiangxi, China.
3. Suijiang Xingbang Agricultural Development Co., Ltd., Zhaotong 657700, Yunnan, China.
4. Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences, Changsha 410205, Hunan, China.
Publication Type:Journal Article
Keywords: Cuniculiplasma divulgatum; GH1 family; enzymatic properties; ethanol tolerance; β-glucosidase
MeSH: beta-Glucosidase; Sodium Chloride; Temperature; Glucose; Ethanol/chemistry*; Ions; Hydrogen-Ion Concentration; Enzyme Stability; Substrate Specificity
From: Chinese Journal of Biotechnology 2023;39(11):4694-4707
CountryChina
Language:Chinese
Abstract: β-glucosidase has important applications in food, pharmaceutics, biomass conversion and other fields, exploring β-glucosidase with strong adaptability and excellent properties thus has received extensive interest. In this study, a novel glucosidase from the GH1 family derived from Cuniculiplasma divulgatum was cloned, expressed, and characterized, aiming to find a better β-glucosidase. The amino acid sequences of GH1 family glucosidase derived from C. divulgatum were obtained from the NCBI database, and a recombinant plasmid pET-30a(+)-CdBglA was constructed. The recombinant protein was induced to express in Escherichia coli BL21(DE3). The enzymatic properties of the purified CdBglA were studied. The molecular weight of the recombinant CdBglA was 56.0 kDa. The optimum pH and temperature were 5.5 and 55 ℃, respectively. The enzyme showed good pH stability, 92.33% of the initial activity could be retained when treated under pH 5.5-11.0 for 1 h. When pNPG was used as a substrate, the kinetic parameters K_m, V_max and K_cat/K_m were 0.81 mmol, 291.99 μmol/(mg·min), and 387.50 s^-1 mmol^-1, respectively. 90.33% of the initial enzyme activity could be retained when CdBglA was placed with various heavy metal ions at a final concentration of 5 mmol/L. The enzyme activity was increased by 28.67% under 15% ethanol solution, remained unchanged under 20% ethanol, and 43.68% of the enzyme activity could still be retained under 30% ethanol. The enzyme has an obvious activation effect at 0-1.5 mol/L NaCl and can tolerate 0.8 mol/L glucose. In conclusion, CdBglA is an acidic and mesophilic enzyme with broad pH stability and strong tolerance to most metal ions, organic solvents, NaCl and glucose. These characteristics may facilitate future theoretical research and industrial production.