ABSTRACT
A novel CYP74 clan gene CYP443С1 of the starlet sea anemone (Nematostella vectensis, Cnidaria) has been cloned, and the properties of the corresponding recombinant protein have been studied. Depending on the substrate, CYP443С1 exhibited double function hydroperoxide lyase/epoxyalcohol synthase activity.
Subject(s)
Aldehyde-Lyases/metabolism , Cytochrome P-450 Enzyme System/metabolism , Sea Anemones/enzymology , Aldehyde-Lyases/chemistry , Aldehyde-Lyases/genetics , Amino Acid Sequence , Animals , Cloning, Molecular , Cytochrome P-450 Enzyme System/chemistry , Cytochrome P-450 Enzyme System/genetics , Sea Anemones/genetics , Sequence AlignmentABSTRACT
Cytochromes P450 of the CYP74 family play a key role in the lipoxygenase cascade generating oxylipins (products of polyunsaturated fatty acid oxidation). The CYP74 family includes allene oxide synthases, hydroperoxide lyases, divinyl ether synthases, and epoxyalcohol synthases. In this work, we cloned the CYP74A88 gene from the Japanese buttercup (Ranunculus japonicus) and studied the properties of the encoded recombinant protein. The CYP74A88 enzyme specifically converts linoleic acid 9- and 13-hydroperoxides to oxiranyl carbinols 9,10-epoxy-11-hydroxy-12-octadecenoic acid and 11-hydroxy-12,13-epoxy-9-octadecenoic acid, respectively, which was confirmed by GC-MS analysis and kinetic studies. Therefore, the CYP74A88 enzyme is a specific epoxyalcohol synthase.
Subject(s)
Biocatalysis , Cloning, Molecular , Cytochrome P-450 Enzyme System/metabolism , Ranunculus/enzymology , Cytochrome P-450 Enzyme System/genetics , Gas Chromatography-Mass Spectrometry , JapanABSTRACT
Data on the influence of the double bond geometry on the antimicrobial properties of different isomers of etherolenic acid against phytopathogenic bacteria are presented. (ω5Z)-Etherolenic acid possesses bactericidal properties against Xanthomonas campestris ssp. vesicatoria, Pseudomonas syringae ssp. tomato, Pectobacterium atrosepticum SCRI1043; the etherolenic and (11Z)-etherolenic acids possess only bacteriostatic properties.
Subject(s)
Anti-Bacterial Agents , Bacteria/growth & development , Fatty Acids, Unsaturated , Lipoxygenase , Plant Proteins , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Fatty Acids, Unsaturated/chemistry , Fatty Acids, Unsaturated/pharmacology , StereoisomerismABSTRACT
The site-directed mutagenesis of tomato allene oxide synthase LeAOS3 (CYP74C3) on the "F/L toggle" site resulted in a partial conversion of LeAOS3 into epoxyalcohol synthase.