ENZYMATIC METHOD FOR DETERMINATION OF BRANCHED-CHAIN AMINO-ACID AMINOTRANSFERASE ACTIVITY

Authors
SCHADEWALDT P HUMMEL W WENDEL U ADELMEYER F
Citation
P. Schadewaldt et al., ENZYMATIC METHOD FOR DETERMINATION OF BRANCHED-CHAIN AMINO-ACID AMINOTRANSFERASE ACTIVITY, Analytical biochemistry, 230(2), 1995, pp. 199-204
Citations number
39
Categorie Soggetti
Biology
Journal title
Analytical biochemistry → ACNP
ISSN journal
00032697
Volume
230
Issue
2
Year of publication
1995
Pages
199 - 204
Database
ISI
SICI code
0003-2697(1995)230:2<199:EMFDOB>2.0.ZU;2-P
Abstract
A spectrophotometric assay for the determination of branched-chain L-a mino acid aminotransferase activity is described. It is based on the t ransamination of L-leucine in the presence of 2-oxoglutarate yielding 4-methyl-2-oxopentanoate. The rate of formation of the branched-chain 2-oxo acid is specifically monitored in a coupled enzymatic reaction u sing NAD(+)-dependent D-2-hydroxyisocaproate dehydrogenase from Lactob acillus casei ssp. pseudoplantarum as coupling enzyme by measuring the decrease in NADH absorbance at 334 nm. Optimized assay conditions are provided as evaluated for the (iso)enzyme from rat heart. (C) 1995 Ac ademic Press, Inc.