Expression of Recombinant <i>β</i>-Glucosidase in <i>Bacillus subtilis</i> and Its Enzymatic Characterization and Application in Preparation of Icariside II

PEI Wenwen; ZENG Yan; LIU Juanjuan; ZHANG Jiangang; WANG Min; SUN Yuanxia

doi:10.13386/j.issn1002-0306.2021080167

Volume 43 Issue 6

Mar. 2022

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Science and Technology of Food Industry > 2022 > 43(6): 133-140. > DOI: 10.13386/j.issn1002-0306.2021080167

PEI Wenwen, ZENG Yan, LIU Juanjuan, et al. Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside II[J]. Science and Technology of Food Industry, 2022, 43(6): 133−140. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021080167.

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Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside II

1.
College of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, China
2.
National Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, China

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Received Date: August 16, 2021
Available Online: January 12, 2022

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Abstract

Abstract

To obtain a safe and efficient β-glucosidase for preparing icariside II from icariin by biocatalytic hydrolysis, a GRAS (generally recognized as safe) strain Bacillus subtilis WB600 was used as the host to produce a thermophilic β-glucosidase from Thermotoga petrophila, by construction of a Escherichia coli-Bacillus subtilis shuttle vector pMA5. Enzymatic properties of the recombinant enzyme and its process conditions of hydrolyzing icariin to prepare icariside II were studied. The result showed that after cultivated in SR culture for 48 h at 30 ℃, the activity of β-glucosidase in the fermented broth reached to 69.68 U/mL. TpBgl3 exhibited the maximal activity at 85 °C and pH4.0. More than 85% of the maximum activity was retained after incubation at 65 °C and pH4.0 for 3 h. Under optimized conditions of 65 °C, pH4.0, and 0.16 U/mg enzyme dosage, the recombinant enzyme could transform 10 g/L icariin into 7.50 g/L icariside II in 20 min with a molar conversion of 98.67%. The successful secretory expression of β-glucosidase in Bacillus subtilis could provide new ways to the industrial safe and efficient biopreparation of icariside II and other high-value-added aglycon compounds.
- β-glucosidase,
- Bacillus subtilis,
- heterologous expression,
- enzymatic properties,
- hydrolysis of icariin,
- icariside II

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References (33)

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