Document Detail


On the specificity of lipid hydroperoxide fragmentation by fatty acid hydroperoxide lyase from Arabidopsis thaliana.
MedLine Citation:
PMID:  12940547     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Fatty acid hydroperoxide lyase (HPL) is a membrane associated P450 enzyme that cleaves fatty acid hydroperoxides into aldehydes and omega-oxo fatty acids. One of the major products of this reaction is (3Z)-hexenal. It is a constituent of many fresh smelling fruit aromas. For its biotechnological production and because of the lack of structural data on the HPL enzyme family, we investigated the mechanistic reasons for the substrate specificity of HPL by using various structural analogues of HPL substrates. To approach this 13-HPL from Arabidopsis thaliana was cloned and expressed in E. coli utilising a His-Tag expression vector. The fusion protein was purified by affinity chromatography from the E. coli membrane fractions and its pH optimum was detected to be pH 7.2. Then, HPL activity against the respective (9S)- and (13S)-hydroperoxides derived either from linoleic, alpha-linolenic or gamma-linolenic acid, respectively, as well as that against the corresponding methyl esters was analysed. Highest enzyme activity was observed with the (13S)-hydroperoxide of alpha-linolenic acid (13alpha-HPOT) followed by that with its methyl ester. Most interestingly, when the hydroperoxy isomers of gamma-linolenic acid were tested as substrates, 9gamma-HPOT and not 13gamma-HPOT was found to be a better substrate of the enzyme. Taken together from these studies on the substrate specificity it is concluded that At13HPL may not recognise the absolute position of the hydroperoxy group within the substrate, but shows highest activities against substrates with a (1Z4S,5E,7Z)-4-hydroperoxy-1,5,7-triene motif. Thus, At13HPL may not only be used for the production of C6-derived volatiles, but depending on the substrate may be further used for the production of Cg-derived volatiles as well.
Authors:
Romy Kandzia; Michael Stumpe; Ekkehardt Berndt; Marlena Szalata; Kenji Matsui; Ivo Feussner
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of plant physiology     Volume:  160     ISSN:  0176-1617     ISO Abbreviation:  J. Plant Physiol.     Publication Date:  2003 Jul 
Date Detail:
Created Date:  2003-08-27     Completed Date:  2003-11-17     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9882059     Medline TA:  J Plant Physiol     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  803-9     Citation Subset:  IM    
Affiliation:
Institute of Plant Biochemistry-Halle/Saale, Weinberg 3, D-06120 Halle/Saale, Germany.
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MeSH Terms
Descriptor/Qualifier:
Aldehyde-Lyases / genetics,  metabolism*
Arabidopsis / enzymology*,  genetics
Cytochrome P-450 Enzyme System / genetics,  metabolism*
Fatty Acids / chemistry,  metabolism
Hydrogen-Ion Concentration
Kinetics
Lipid Peroxides / chemistry,  metabolism*
Molecular Structure
Substrate Specificity
Chemical
Reg. No./Substance:
0/Fatty Acids; 0/Lipid Peroxides; 9035-51-2/Cytochrome P-450 Enzyme System; EC 4.1.2.-/Aldehyde-Lyases; EC 4.1.2.-/hydroperoxide lyase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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