TITLE

A new cytochrome P450 system from Bacillus megaterium DSM319 for the hydroxylation of 11-keto-β-boswellic acid (KBA)

AUTHOR(S)
Brill, Elisa; Hannemann, Frank; Zapp, Josef; Brüning, Gerit; Jauch, Johann; Bernhardt, Rita
PUB. DATE
February 2014
SOURCE
Applied Microbiology & Biotechnology;Feb2014, Vol. 98 Issue 4, p1703
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
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 11-keto-β-boswellic 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 whole-cell biocatalysts were constructed based on B. megaterium MS941. The main product has been identified as 7β-hydroxy-KBA, while the side product (∼20 %) was shown to be a mixture of 7β,15α-dihydroxy-KBA and 15α-hydroxy-KBA. Without further optimization 560.7 mg l day of the main product, 7β-hydroxy-KBA, 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.
ACCESSION #
94355453

 

Related Articles

  • Laboratory evolution of a soluble, self-sufficient, highly active alkane hydroxylase. Glieder, Anton; Farinas, Edgardo T.; Arnold, Frances H. // Nature Biotechnology;Nov2002, Vol. 20 Issue 11, p1135 

    We have converted cytochrome P450 BM-3 from Bacillus megaterium (P450 BM-3), a medium-chain (C12-C18) fatty acid monooxygenase, into a highly efficient catalyst for the conversion of alkanes to alcohols. The evolved P450 BM-3 exhibits higher turnover rates than any reported biocatalyst for the...

  • A new Bacillus megaterium whole-cell catalyst for the hydroxylation of the pentacyclic triterpene 11-keto-β-boswellic acid (KBA) based on a recombinant cytochrome P450 system. Bleif, Sabrina; Hannemann, Frank; Zapp, Josef; Hartmann, David; Jauch, Johann; Bernhardt, Rita // Applied Microbiology & Biotechnology;Feb2012, Vol. 93 Issue 3, p1135 

    The use of cytochromes P450 for the regio- and stereoselective hydroxylation of non-activated carbon atoms in biotechnological applications reflects an efficient and cost-effective alternative in comparison to classical organic chemistry. The prokaryotic cytochrome P450 CYP106A2 from Bacillus...

  • Covalent immobilization of cytochrome P450 BM3 (R966D/W1046S) on glutaraldehyde activated SPIONs. Bahrami, Atieh; Garnier, Alain; Larachi, Faïçal; Iliuta, Maria C. // Canadian Journal of Chemical Engineering;Oct2018, Vol. 96 Issue 10, p2227 

    Abstract: Selective hydroxylation of the C‐H bond of saturated hydrocarbon chains at room temperature is the signature of an invaluable biocatalyst, cytochrome P450 BM3 from Bacillus megaterium. Despite this remarkable ability, because of the enzyme's inherent low stability and dependence...

  • Metabolism of polychlorinated dibenzo-p-dioxins by cytochrome P450 BM-3 and its mutant. Woro Sulistyaningdyah; Jun Ogawa; Qing-Shan Li; Raku Shinkyo; Toshiyuki Sakaki; Inouye, Kuniyo; Schmid, Rolf; Sakayu Shimizu // Biotechnology Letters;Dec2004, Vol. 26 Issue 24, p1857 

    The metabolism of polychlorinated dibenzo-p-dioxins by cytochrome P450 BM-3 fromBacillus megateriumand a mutant enzyme of it (AL4V; Ala74Gly, Phe87Val, Leu188Gln triple mutant) was examined. Both purified enzymes metabolized 1-monochloro-, 2,3-dichloro-, and 2,3,7-trichloro-dibenzo-p-dioxin, but...

  • Cytochrome P450foxy, a Catalytically Self-Sufficient Fatty Acid Hydroxylase of the Fungus Fusarium oxysporum1. Nakayama, Norikazu; Takemae, Asako; Shoun, Hirofumi // Journal of Biochemistry;1996, Vol. 119 Issue 3, p435 

    We have purified membrane-bound fatty acid (ω-1-ω-3) hydroxylase of the fungus Fusarium oxysporum MT-811 and found that the activity depends on a single polypeptide with an apparent Mr value of 118,000. The purified hydroxylase exhibited spectral characteristics of cytochrome P450...

  • Catalytic activity, duplication and evolution of the CYP98 cytochrome P450 family in wheat. Pascaline Ullmann; Tanya Ertunç; Dawn Little; Carl Olsen; Maike Petersen; Jonathan Negrel; Danièle Werck-Reichhart // Plant Molecular Biology;Jan2007, Vol. 63 Issue 1, p1 

    Abstract??A burst of evolutionary duplication upon land colonization seems to have led to the large superfamily of cytochromes P450 in higher plants. Within this superfamily some clans and families are heavily duplicated. Others, such as genes involved in the phenylpropanoid pathway have led to...

  • Reconstitution of monooxygenase activity of membrane cytochromes P450 in vitro and detergents. Myasoedova, K.; Arutyunyan, A.; Magretova, N. // Doklady Biochemistry & Biophysics;Jul2007, Vol. 415 Issue 1, p174 

    The article focuses on the reconstitution of monooxygenase activity of membrane cytochromes P450 (CYPs) in vitro and detergents. The catalytic function of these hemoproteins is hydroxylation of hydrophobic molecules in monooxygenase reactions. The catalytic activity of membrane CYPs is studied...

  • Oxidation of dibenzo-p-dioxin, dibenzofuran, biphenyl, and diphenyl ether by the white-rot fungus Phlebia lindtneri. Mori, T.; Kondo, R. // Applied Microbiology & Biotechnology;Oct2002, Vol. 60 Issue 1/2, p200 

    Hydroxylation of dibenzo-p-dioxin (DD), dibenzofuran (DF), biphenyl (BP) and diphenyl ether (DPE) by the white-rot fungus Phlebia lindtneri GB-1027 was studied. DD and DF were rapidly degraded in a culture of P. lindtneri. The initial oxidation products were identified by gas chromatography-mass...

  • Laboratory evolution of peroxide-mediated cytochrome P450 hydroxylation. Joo, Hyun; Lin, Zhanglin; Arnold, Frances H. // Nature;6/17/1999, Vol. 399 Issue 6737, p670 

    Reports the directed evolution of cytochrome P450 monooxygenases (P450s) from Pseudomonas putida to create mutants that hydroxylate naphthalene in the absence of cofactors through the peroxide shunt pathway. Ability to screen efficiently for improved mutants; Ability to select and develop mono...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics