| A high-performance liquid chromatography-based radiometric assay for acyl-CoA:alcohol transacylase from jojoba. | |
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MedLine Citation:
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PMID: 1481989 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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W S Garver; J D Kemp; G D Kuehn |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Analytical biochemistry Volume: 207 ISSN: 0003-2697 ISO Abbreviation: Anal. Biochem. Publication Date: 1992 Dec |
Date Detail:
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Created Date: 1993-02-11 Completed Date: 1993-02-11 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0370535 Medline TA: Anal Biochem Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 335-40 Citation Subset: IM |
Affiliation:
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Plant Genetic Engineering Laboratory, New Mexico State University, Department 3C, Las Cruces 88003-0001. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Acyl Coenzyme A
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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:
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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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