Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside II
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 Escheric...
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The editorial department of Science and Technology of Food Industry
2022-03-01
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| Series: | Shipin gongye ke-ji |
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| Online Access: | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021080167 |
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| author | Wenwen PEI Yan ZENG Juanjuan LIU Jiangang ZHANG Min WANG Yuanxia SUN |
| author_facet | Wenwen PEI Yan ZENG Juanjuan LIU Jiangang ZHANG Min WANG Yuanxia SUN |
| author_sort | Wenwen PEI |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T09:09:15Z |
| format | Article |
| id | doaj.art-8d99f9983fe342b58a098ea8b588b7b8 |
| institution | Directory Open Access Journal |
| issn | 1002-0306 |
| language | zho |
| last_indexed | 2024-04-13T09:09:15Z |
| publishDate | 2022-03-01 |
| publisher | The editorial department of Science and Technology of Food Industry |
| record_format | Article |
| series | Shipin gongye ke-ji |
| spelling | doaj.art-8d99f9983fe342b58a098ea8b588b7b82022-12-22T02:52:56ZzhoThe editorial department of Science and Technology of Food IndustryShipin gongye ke-ji1002-03062022-03-0143613314010.13386/j.issn1002-0306.20210801672021080167-6Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside IIWenwen PEI0Yan ZENG1Juanjuan LIU2Jiangang ZHANG3Min WANG4Yuanxia SUN5College of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, ChinaNational Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, ChinaNational Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, ChinaCollege of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, ChinaCollege of Bioengineering, Tianjin University of Science and Technology, Tianjin 300457, ChinaNational Engineering Laboratory for Industrial Enzymes, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin 300308, ChinaTo 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.http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021080167β-glucosidasebacillus subtilisheterologous expressionenzymatic propertieshydrolysis of icariinicariside ii |
| spellingShingle | Wenwen PEI Yan ZENG Juanjuan LIU Jiangang ZHANG Min WANG Yuanxia SUN Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside II Shipin gongye ke-ji β-glucosidase bacillus subtilis heterologous expression enzymatic properties hydrolysis of icariin icariside ii |
| title | Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside II |
| title_full | Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside II |
| title_fullStr | Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside II |
| title_full_unstemmed | Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside II |
| title_short | Expression of Recombinant β-Glucosidase in Bacillus subtilis and Its Enzymatic Characterization and Application in Preparation of Icariside II |
| title_sort | expression of recombinant β glucosidase in bacillus subtilis and its enzymatic characterization and application in preparation of icariside ii |
| topic | β-glucosidase bacillus subtilis heterologous expression enzymatic properties hydrolysis of icariin icariside ii |
| url | http://www.spgykj.com/cn/article/doi/10.13386/j.issn1002-0306.2021080167 |
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