Journal of Applied Biological Chemistry
- Volume 43 Issue 4
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- Pages.230-234
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- 2000
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- 1976-0442(pISSN)
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- 2234-7941(eISSN)
Simple Assay Method for Determination of Capsaicinoid Synthetase Activity
- Kim, Kye-Won (Division of Biochemistry, National Institute of Agricultural Science and Technology) ;
- Varindra, R. (Department of Biochemistry, Punjab Agricultural University) ;
- Kim, Donghern (Division of Biochemistry, National Institute of Agricultural Science and Technology) ;
- Hwang, Seon-Kap (Department of Biochemistry, Punjab Agricultural University) ;
- Kim, Jong-Guk (Department of Microbiology, Kyungpook National University) ;
- Lee, Shin-Woo (Division of Biochemistry, National Institute of Agricultural Science and Technology)
- Received : 2000.09.02
- Published : 2000.12.31
Abstract
A new method to assay the capsaicinoid synthetase (CS) activity was developed by utilizing NADHcoupled enzyme systems involving pyruvate kinase and lactate dehydrogenase. CS activities in Capsicum placenta, depending upon the kinetics of the NADH oxidation, revealed almost the same profile as compared with those shown using an HPLC-based method. When the substrates, 8-methyl nonanoic acid and vanillylamine, for the CS enzyme were employed separately or simultaneously, it appeared that the two-step reaction, acyl-CoA formation and condensation with vanillyla~ne, of the CS enzyme was a coupled reaction. Thus, this assay method of the CS enzyme can be considered as an alternative to the HPLC-based method, since it has the advantages of rapidity and simplicity as well as reliability when compared with the existing method.