Glutamate-branched-chain amino acid transaminase

摘要: Publisher Summary Branched-chain amino acid transaminase activity is widespread. The supernatant pig heart enzyme active with at least eleven acids but it most the branched-chain acids. various in order: isoleucine > valine DL-alloisoleucine leucine. assay procedures for transaminases involve selective extraction of α-keto 2,4-dinitrophenylhydrazone into a suitable organic solvent. In one methods used, reaction mixture contains 20 μmol L-leucine, α-ketoglutarate, 200 Tris (pH 8.6), and final volume 3.0 ml, 37 ° . After incubation 10 minutes, terminated by addition 0.3% 2, 4-dinitrophenylhydrazine 2 M HCl. phases are separated brief centrifugation clinical centrifuge. upper phase removed α-ketoisocaproate-2,4-dinitrophenylhydrazone back extracted 1.5 ml 10% w/v sodium carbonate.