Document Detail


A high-performance liquid chromatography-based radiometric assay for acyl-CoA:alcohol transacylase from jojoba.
MedLine Citation:
PMID:  1481989     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Acyl-CoA:alcohol transacylase catalyzes the final step in the biosynthesis of storage liquid wax esters from acyl-CoA fatty acids and fatty alcohols in a limited number of microbes, algae, and Simmondsia chinensis Link (jojoba). An improved and automated method of enzyme assay for this catalyst from cotyledons of jojoba is described. The assay method uses reversed-phase C18 high performance liquid chromatography (HPLC) to separate the labeled C30:1 liquid wax product, [14C]-dodecanyl-octadecenoate, from the unreacted substrate, [14C]octadecenoyl-CoA (oleyl-CoA), and other components produced from enzymes present in the crude homogenate of jojoba cotyledons, including [14C]-octadecenoic acid (oleic acid) and [14C]octadecenol (oleyol). Methods are also described for microscale chemical synthesis in one vessel of 14C-radiolabeled substrates and products for the transacylase. These labeled reagents are required to confirm the HPLC separations of reaction products. The radioactive components are quantitated using an on-line flow-through scintillation detector enabling sensitive and precise analysis of the reaction products.
Authors:
W S Garver; J D Kemp; G D Kuehn
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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:  Analytical biochemistry     Volume:  207     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  1992 Dec 
Date Detail:
Created Date:  1993-02-11     Completed Date:  1993-02-11     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  335-40     Citation Subset:  IM    
Affiliation:
Plant Genetic Engineering Laboratory, New Mexico State University, Department 3C, Las Cruces 88003-0001.
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MeSH Terms
Descriptor/Qualifier:
Acyl Coenzyme A / isolation & purification,  metabolism
Acyltransferases / analysis*,  metabolism
Carbon Radioisotopes
Chromatography, High Pressure Liquid / methods
Isotope Labeling / methods
Kinetics
Oleic Acids / isolation & purification*
Plants / enzymology*
Radioisotope Dilution Technique
Thermodynamics
Chemical
Reg. No./Substance:
0/Acyl Coenzyme A; 0/Carbon Radioisotopes; 0/Oleic Acids; 1716-06-9/oleoyl-coenzyme A; 36078-10-1/dodecyl oleate; EC 2.3.-/Acyltransferases; EC 2.3.1.75/long-chain-alcohol O-fatty-acyltransferase

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


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