Matthias Dietrich

Publications (6)18.28 Total impact

• Article: Identification and functional expression of △9 fatty acid desaturase from the marine bacterium Pseudoalteromonas sp. MLY15

  Yan Li, Xian Xu, Matthias Dietrich, Vlada B. Urlacher, Rolf D. Schmid, Pingkai Ouyang, Bingfang He
  Journal of Molecular Catalysis B Enzymatic 01/2009; · 2.73 Impact Factor
• Article: Altering the regioselectivity of the subterminal fatty acid hydroxylase P450 BM-3 towards gamma- and delta-positions.

  Matthias Dietrich, Tuan Anh Do, Rolf D Schmid, Jürgen Pleiss, Vlada B Urlacher
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  ABSTRACT: Cytochrome P450 BM-3 monooxygenase from Bacillus megaterium (CYP102A1) catalyzes the subterminal hydroxylation of fatty acids with a chain length of 12-22 carbons. Wild-type P450 BM-3 oxidizes saturated fatty acids at subterminal positions producing a mixture of omega-1, omega-2 and omega-3 hydroxylated products. Using a rational site-directed mutagenesis approach, three new elements have been introduced into the substrate binding pocket of the monooxygenase, which greatly changed the product pattern of lauric acid hydroxylation. Particularly, substitutions at positions S72, V78 and I263 had an effect on the enzyme regioselectivity. The P450 BM-3 mutants V78A F87A I263G and S72Y V78A F87A were able to oxidize lauric acid not only at delta-position (14% and 16%, respectively), but also produced gamma- and beta-hydroxylated products. delta-Hydroxy lauric and gamma-hydroxy lauric acid are important synthons for the production of the corresponding lactones.
  Journal of Biotechnology 11/2008; 139(1):115-7. · 3.05 Impact Factor
• Article: Identification and functional expression of a Delta9-fatty acid desaturase from Psychrobacter urativorans in Escherichia coli.

  Yan Li, Matthias Dietrich, Rolf D Schmid, Bingfang He, Pingkai Ouyang, Vlada B Urlacher
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  ABSTRACT: The Delta9-fatty acid desaturase is a key enzyme in the synthesis of unsaturated fatty acids. The fatty acid composition of membrane phospholipids in Psychrobacter urativorans is characterized by a high degree of desaturation at Delta9 position. Based on CODEHOP-mediated PCR strategy, a novel gene designated as PuFAD9, putatively encoding a Delta9-fatty acid desaturase (PuFAD9), was isolated from P. urativorans. The gene consists of 1,455 bp and codes for 484 amino acids. Analysis of the amino acid sequence reveals three histidine clusters and a hydropathy profile, typical for membrane-bound desaturases. Activity of the PuFAD9 protein, recombinantly expressed in Escherichia coli was confirmed by GC-MS analysis of the cellular fatty acid composition. It was found that the ratio between palmitoleic and palmitic acid in E. coli cells heterologously expressing the PuFAD9 gene was significantly affected by IPTG induction and the growth temperature.
  Lipids 04/2008; 43(3):207-13. · 2.13 Impact Factor
• Article: Cytochrome P450 monooxygenase from Clostridium acetobutylicum: a new alpha-fatty acid hydroxylase.

  Marco Girhard, Stefanie Schuster, Matthias Dietrich, Peter Dürre, Vlada B Urlacher
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  ABSTRACT: Cytochrome P450 monooxygenase from the anaerobic microorganism Clostridium acetobutylicum (CYP152A2) has been produced in Escherichia coli. CYP152A2 was shown to bind a broad range of saturated and unsaturated fatty acids and corresponding methyl esters and demonstrated a high peroxygenase activity of up to 200min(-1) with myristic acid. Although a high concentration of hydrogen peroxide of 200microM was necessary for high activities of the enzyme, it led to a fast enzyme inactivation within 2-4min. This might reflect the natural function of CYP152A2 as a rapid hydrogen peroxide scavenging enzyme. In two different reconstituted systems with NADPH, CYP152A2 was able to convert 10 times more substrate, if provided with flavodoxin and flavodoxin reductase from E. coli and even 30-40 times more substrate with the CYP102A1-reductase from Bacillus megaterium. According to the clear preference for hydroxylation at alpha-position, CYP152A2 can be referred to as fatty acid alpha-hydroxylase.
  Biochemical and Biophysical Research Communications 11/2007; 362(1):114-9. · 2.48 Impact Factor
• Source

  Available from: uni-stuttgart.de

  Article: Recombinant production of human microsomal cytochrome P450 2D6 in the methylotrophic yeast Pichia pastoris.

  Matthias Dietrich, Lisa Grundmann, Katja Kurr, Laura Valinotto, Tanja Saussele, Rolf D Schmid, Stefan Lange
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  ABSTRACT: Microsomal cytochrome P450 monooxygenases of groups 1-3 are mainly expressed in the liver and play a crucial role in phase 1 reactions of xenobiotic metabolism. The cDNAs encoding human CYP2D6 and human NADPH-P450 oxidoreductase (CPR) were transformed into the methylotrophic yeast Pichia pastoris and expressed with control of the methanol-inducible AOX1 promoter. The determined molecular weights of the recombinant CYP2D6 and CPR closely matched the calculated values of 55.8 and 76.6 kDa. CPR activity was detected by conversion of cytochrome c by using isolated microsomes. Nearly all of the recombinant CYP was composed of the active holoenzyme, as confirmed by reduced CO difference spectra, which showed a single peak at 450 nm. Only by coexpression of human CPR and CYP was CYP2D6 activity obtained. Microsomes containing human CPR and CYP2D6 converted different substrates, such as 3-cyano-7-ethoxycoumarin, parathion and dextrometorphan. The kinetic parameters of dextrometorphan conversion closely matched those of CYP2D6 from other recombinant expression systems and human microsomes. The endogenous NADPH-P450 oxidoreductase of Pichia pastoris seems to be incompatible with human CYP2D6, as expression of CYP2D6 without human CPR did not result in any CYP activity. These recombinant strains provide a novel, easy-to-handle and cheap source for the biochemical characterisation of single microsomal cytochromes, as well as their allelic variants.
  ChemBioChem 12/2005; 6(11):2014-22. · 3.94 Impact Factor
• Article: Recombinant Production of Human Microsomal Cytochrome P450 2D6 in the Methylotrophic Yeast Pichia pastoris

  Matthias Dietrich, Lisa Grundmann, Katja Kurr, Laura Valinotto, Tanja Saussele Dr, Rolf D. Schmid Prof. Dr, Stefan Lange Dr
  ChemBioChem 10/2005; 6(11):2014 - 2022. · 3.94 Impact Factor