Title:
PREPARATION METHOD FOR DEHYDROEPIANDROSTERONE, AND ENZYME FOR PREPARATION THEREOF
Document Type and Number:
WIPO Patent Application WO/2019/010971
Kind Code:
A1
Abstract:
A preparation method for dehydroepiandrosterone, comprising: in a protective atmosphere, adding potassium tert-butoxide to tert-butanol, stirring evenly, adding 4-androstenedione to obtain a mixture, and adding the mixture dropwise to a sodium ascorbate-containing acetic acid solution for a reaction to obtain 5-androstenedione; dissolving the 5-androstenedione in an organic solvent, adding a ketone reductase, a glucose dehydrogenase, glucose and a redox coenzyme to obtain a mixture, controlling the pH of the mixture to be 6.0-6.3, stirring and reacting for 1-6 hours at 22-26°C to obtain a reaction solution, and performing separation and purification on the reaction solution to obtain dehydroepiandrosterone, the ketone reductase and the glucose dehydrogenase being coexpressed by a microbial strain and added in the form of a crude enzyme solution. The synthesis process of the preparation method has few steps, simple operations, high yields and low costs, and may be widely applied to industrial scale production. Also provided is an enzyme for preparation.
Inventors:
FU RONGZHAO (CN)
LIU LIHUI (CN)
CHEN XIAOCHUN (CN)
CAO LEI (CN)
LIU TAOTAO (CN)
LIU LIHUI (CN)
CHEN XIAOCHUN (CN)
CAO LEI (CN)
LIU TAOTAO (CN)
Application Number:
PCT/CN2018/075822
Publication Date:
January 17, 2019
Filing Date:
February 08, 2018
Export Citation:
Assignee:
BONTAC BIO ENG SHENZHEN CO LTD (CN)
International Classes:
C12P33/06; C07K14/195; C12N9/02; C12N15/53; C12N15/64
Foreign References:
| CN106086148A | 2016-11-09 | |||
| CN106636285A | 2017-05-10 | |||
| CN105695551A | 2016-06-22 |
Other References:
DATABASE GenBank 22 January 2016 (2016-01-22), SAHL, J. ET AL.: "3-alpha-hydroxysteroid dehydrogenase[Burkholderia territorii", Database accession no. KWA22726. 1
DATABASE G enBank 22 January 2016 (2016-01-22), SAHL, J. ET AL.: "3-alpha-hydroxysteroid dehydrogenase[Burkholderia territorii", Database accession no. KWA22726. 1.
DATABASE GenBank 17 April 2017 (2017-04-17), ANASTASI, E. ET AL.: "3-alpha-hydroxysteroid dehydrogenase[Rhodococcus hoagii", Database accession no. ORI1)9999 . 1
DATABASE GenBank 17 April 2017 (2017-04-17), ANASTASI, E. ET AL.: "3-alpha-hydroxysteroid dehydrogenase[Rhodococcus hoagii", Database accession no. ORL09999. 1
DATABASE GenBank 9 December 2016 (2016-12-09), KANTOR, R.S. ET AL.: "3-alpha-hydroxysteroid dehydrogenase[Sphingomonas sp.67-36", Database accession no. OJV28933.1
DATABASE G enBank 22 January 2016 (2016-01-22), SAHL, J. ET AL.: "3-alpha-hydroxysteroid dehydrogenase[Burkholderia territorii", Database accession no. KWA22726. 1.
DATABASE GenBank 17 April 2017 (2017-04-17), ANASTASI, E. ET AL.: "3-alpha-hydroxysteroid dehydrogenase[Rhodococcus hoagii", Database accession no. ORI1)9999 . 1
DATABASE GenBank 17 April 2017 (2017-04-17), ANASTASI, E. ET AL.: "3-alpha-hydroxysteroid dehydrogenase[Rhodococcus hoagii", Database accession no. ORL09999. 1
DATABASE GenBank 9 December 2016 (2016-12-09), KANTOR, R.S. ET AL.: "3-alpha-hydroxysteroid dehydrogenase[Sphingomonas sp.67-36", Database accession no. OJV28933.1
Attorney, Agent or Firm:
SCIHEAD IP LAW FIRM (CN)
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