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Biotechnological Approach to the Synthesis of 9α-Hydroxylated Steroids
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Physical and Analytical Chemistry.
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2007 (English)In: Preparative Biochemistry & Biotechnology, ISSN 1082-6068, E-ISSN 1532-2297, Vol. 37, no 4, p. 309-321Article in journal (Refereed) Published
Abstract [en]

The steroid 9α-hydroxylase gene has been cloned from Mycobacterium smegmatis into Escherichia coli BL21. Progesterone added to bioreactors was subjected to in vivo transformation into 9α-hydroxyprogesterone. In 7 days, 43.6 mg

9α-hydroxyprogesterone was formed from 53.8 mg/L progesterone. The enzyme also has shown evidence of processing 4-androstene-3,17-dione in vivo. An extensive analytical method development, including LLE, HPLC-DAD, MS, and

NMR was performed to verify the product and to enable a quantitative analysis. Protocols for analytical and preparative separation have been developed, using binaphtol as internal standard. Both the growth pattern and the bioconversion rate

were unaffected by the presence of binaphtol in the bioreactor. The enzyme was purified by immobilised metal affinity and ion exchange chromatography, resulting in low in vitro activity.

Place, publisher, year, edition, pages
2007. Vol. 37, no 4, p. 309-321
Keyword [en]
Progesterone, 4-Androstene-3, 17-dione, Steroid, 9α-Hydroxylation, 9α-Hydroxyprogesterone, Recombinant expression
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:uu:diva-12318DOI: 10.1080/10826060701593209ISI: 000249781000001OAI: oai:DiVA.org:uu-12318DiVA, id: diva2:40087
Available from: 2007-12-11 Created: 2007-12-11 Last updated: 2018-01-23Bibliographically approved

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Fornstedt, TorgnyErsson, Bo

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