Cytochrome P450 monooxygenase from Clostridium acetobutylicum: a new alpha-fatty acid hydroxylase.
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.
Girhard
Marco
M
Institute of Technical Biochemistry, University of Stuttgart, 70569 Stuttgart, Germany.
Schuster
Stefanie
S
Institute of Microbiology and Biotechnology, University of Ulm, 89069 Ulm, Germany.
Dietrich
Matthias
M
Institute of Technical Biochemistry, University of Stuttgart, 70569 Stuttgart, Germany.
Dürre
Peter
P
Institute of Microbiology and Biotechnology, University of Ulm, 89069 Ulm, Germany.
Urlacher
Vlada B
VB
Institute of Technical Biochemistry, University of Stuttgart, 70569 Stuttgart, Germany. Electronic address: Vlada.Urlacher@itb.uni-stuttgart.de.
D001426:Bacterial Proteins / Q000378:metabolism; D001665:Binding Sites; D003001:Cloning, Molecular; D046969:Clostridium acetobutylicum / Q000201:enzymology*; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology*; D005227:Fatty Acids / Q000737:chemistry / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000378:metabolism; D008954:Models, Biological; D008956:Models, Chemical; D009251:NADPH-Ferrihemoprotein Reductase; D011485:Protein Binding; D013379:Substrate Specificity
D001426:Bacterial Proteins; D005227:Fatty Acids; D003577:Cytochrome P-450 Enzyme System; D006861:Hydrogen Peroxide; D006899:Mixed Function Oxygenases; C022333:peroxygenase; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases