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Cloning and Characterization of β-Fructosidase from Thermotoga neapolitana DSM4359 and Through the Degraded Glucoside

Author:
Bi, Yun-feng, Xu, Lin-lin, Chen, Ping, Jiang, Shan, Shen, Ming-hao
Source:
Proceedings of the National Academy of Sciences, India, Section B: biological sciences 2018 v.88 no.4 pp. 1499-1505
ISSN:
0369-8211
Subject:
Escherichia coli, Thermotoga neapolitana, beta-fructofuranosidase, enzyme activity, genes, glucosides, inulin, kestose, magnesium, pH, polyacrylamide gel electrophoresis, sucrose, syrups, temperature, thermal stability
Abstract:
The gene of β-fructosidase from Thermotoga neapolitana DMS4359 was cloned and expressed in Escherichia coli. The molecule weight of β-fructosidase detected by SDS-PAGE is 60.0 kDa and the enzyme activity is 6991.33 U/mL. The results of the enzyme characteristics are obtained as the optimal temperature 85 °C, the optimal pH 5.0, and the best promoter 10 mmol/L of Mg²⁺. β-Fructosidase have high thermostability and retained >50% activity after incubated for 40 h at 95 °C. Research on degraded glycosides showed that β-fructosidase can act on sucrose and inulin to produce kestose and fructose–glucose syrup. All these findings confirmed that β-fructosidase can tolerate high temperature and can be widely used in industrial production.
Agid:
6195396