Document Detail


Molecular analysis of a bifunctional fatty acid conjugase/desaturase from tung. Implications for the evolution of plant fatty acid diversity.
MedLine Citation:
PMID:  12481086     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The seed oil derived from the tung (Aleurites fordii Hemsl.) tree contains approximately 80% alpha-eleostearic acid (18:3delta(9cis,11trans,13trans)), an unusual conjugated fatty acid that imparts industrially important drying qualities to tung oil. Here, we describe the cloning and functional analysis of two closely related Delta(12) oleate desaturase-like enzymes that constitute consecutive steps in the biosynthetic pathway of eleostearic acid. Polymerase chain reaction screening of a tung seed cDNA library using degenerate oligonucleotide primers resulted in identification of two desaturases, FAD2 and FADX, that shared 73% amino acid identity. Both enzymes were localized to the endoplasmic reticulum of tobacco (Nicotiana tabacum cv Bright-Yellow 2) cells, and reverse transcriptase-polymerase chain reaction revealed that FADX was expressed exclusively within developing tung seeds. Expression of the cDNAs encoding these enzymes in yeast (Saccharomyces cerevisiae) revealed that FAD2 converted oleic acid (18:1delta(9cis)) into linoleic acid (18:2delta(9cis,12cis)) and that FADX converted linoleic acid into alpha-eleostearic acid. Additional characterization revealed that FADX exhibited remarkable enzymatic plasticity, capable of generating a variety of alternative conjugated and delta(12)-desaturated fatty acid products in yeast cells cultured in the presence of exogenously supplied fatty acid substrates. Unlike other desaturases reported to date, the double bond introduced by FADX during fatty acid desaturation was in the trans, rather than cis, configuration. Phylogenetic analysis revealed that tung FADX is grouped with delta(12) fatty acid desaturases and hydroxylases rather than conjugases, which is consistent with its desaturase activity. Comparison of FADX and other lipid-modifying enzymes (desaturase, hydroxylase, epoxygenase, acetylenase, and conjugase) revealed several amino acid positions near the active site that may be important determinants of enzymatic activity.
Authors:
John M Dyer; Dorselyn C Chapital; Jui-Chang W Kuan; Robert T Mullen; Charlotta Turner; Thomas A McKeon; Armand B Pepperman
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Plant physiology     Volume:  130     ISSN:  0032-0889     ISO Abbreviation:  Plant Physiol.     Publication Date:  2002 Dec 
Date Detail:
Created Date:  2002-12-13     Completed Date:  2003-05-30     Revised Date:  2010-09-14    
Medline Journal Info:
Nlm Unique ID:  0401224     Medline TA:  Plant Physiol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2027-38     Citation Subset:  IM    
Affiliation:
United States Department of Agriculture-Agricultural Research Service Southern Regional Research Center, 1100 Robert E Lee Boulevard, New Orleans, Louisiana 70124, USA. jdyer@nola.srrc.usda.gov
Data Bank Information
Bank Name/Acc. No.:
GENBANK/AF525534;  AF525535
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MeSH Terms
Descriptor/Qualifier:
Aleurites / enzymology*,  genetics
Amino Acid Sequence
Evolution, Molecular
Fatty Acid Desaturases / genetics*,  metabolism
Fatty Acids / chemistry,  metabolism*
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Plant
Mass Spectrometry
Molecular Sequence Data
Phylogeny
Plant Proteins / genetics,  metabolism
Saccharomyces cerevisiae / genetics,  metabolism
Seeds / growth & development
Sequence Homology, Amino Acid
Substrate Specificity
Chemical
Reg. No./Substance:
0/Fatty Acids; 0/Plant Proteins; EC 1.14.19.-/Fatty Acid Desaturases
Comments/Corrections

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


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