| Multi-enzymatic one-pot reduction of dehydrocholic acid to 12-keto-ursodeoxycholic acid with whole-cell biocatalysts. | |
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MedLine Citation:
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PMID: 22806613 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Ursodeoxycholic acid (UDCA) is a bile acid of industrial interest as it is used as an agent for the treatment of primary sclerosing cholangitis and the medicamentous, non-surgical dissolution of gallstones. Currently, it is prepared industrially from cholic acid following a seven-step chemical procedure with an overall yield of less than 30%. In this study, we investigated the key enzymatic steps in the chemo-enzymatic preparation of UDCA - the two-step reduction of dehydrocholic acid (DHCA) to 12-keto-ursodeoxycholic acid using a mutant of 7β-hydroxysteroid dehydrogenase (7β-HSDH) from Collinsella aerofaciens and 3α-hydroxysteroid dehydrogenase (3α-HSDH) from Comamonas testosteroni. Three different one-pot reaction approaches were investigated using whole-cell biocatalysts in simple batch processes. We applied one-biocatalyst systems, where 3α-HSDH, 7β-HSDH and either a mutant of formate dehydrogenase (FDH) from Mycobacterium vaccae N10 or a glucose dehydrogenase (GDH) from Bacillus subtilis were expressed in a Escherichia coli BL21(DE3) based host strain. We also investigated two-biocatalyst systems, where 3α-HSDH and 7β-HSDH were expressed separately together with FDH enzymes for cofactor regeneration in two distinct E. coli hosts that were simultaneously applied in the one-pot reaction. The best result was achieved by the one-biocatalyst system with GDH for cofactor regeneration, which was able to completely convert 100 mM DHCA to > 99.5 mM 12-keto-UDCA within 4.5 h in a simple batch process on a liter scale. Biotechnol. Bioeng. © 2012 Wiley Periodicals, Inc. |
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Authors:
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Boqiao Sun; Christina Kantzow; Sven Bresch; Kathrin Hölsch; Dirk Weuster-Botz |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2012-7-13 |
Journal Detail:
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Title: Biotechnology and bioengineering Volume: - ISSN: 1097-0290 ISO Abbreviation: - Publication Date: 2012 Jul |
Date Detail:
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Created Date: 2012-7-18 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 7502021 Medline TA: Biotechnol Bioeng Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2012 Wiley Periodicals, Inc. |
Affiliation:
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Institute of Biochemical Engineering, Technische Universität München, Boltzmannstr. 15, 85748 Garching, Germany. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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