Authors: Elisa Brill Frank Hannemann Josef Zapp Gerit Brüning Johann Jauch Rita Bernhardt
Publish Date: 2013/06/25
Volume: 98, Issue: 4, Pages: 1703-1717
Abstract
In the genome of Bacillus megaterium DSM319 a strain who has recently been sequenced to fully exploit its potential for biotechnological purposes we identified a gene encoding the cytochrome P450 CYP106A1 as well as genes encoding potential redox partners of CYP106A1 We cloned expressed and purified CYP106A1 and five potential autologous redox partners one flavodoxin and four ferredoxins The flavodoxin and three ferredoxins were able to support the activity of CYP106A1 displaying the first cloned natural redox partners of a cytochrome P450 from B megaterium The CYP106A1 system was able to convert the pentacyclic triterpene 11ketoβboswellic acid KBA belonging to the main bioactive constituents of Boswellia serrata gum resin extracts which are used to treat inflammatory disorders and arthritic diseases In order to provide sufficient amounts of the KBA products to characterize them structurally by NMR spectroscopy recombinant wholecell biocatalysts were constructed based on B megaterium MS941 The main product has been identified as 7βhydroxyKBA while the side product ∼20 was shown to be a mixture of 7β15αdihydroxyKBA and 15αhydroxyKBA Without further optimization 5607 mg l−1 day−1 of the main product 7βhydroxyKBA could be obtained thus providing a suitable starting point for the efficient production of modified KBA by chemical tailoring to produce novel KBA derivatives with increased bioavailability and this way more efficient drugs
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