[A spectrophotometric method for determining the concentration of L-lysine using L-lysine-alpha-oxidase from Trichoderma sp. and 3,3',5,5'-tetramethylbenzidene dihydrochloride].
A simple and relatively sensitive procedure was developed for determination of L-lysine at 3-30 mmole/L concentration. The procedure does not involve the carcinogenic compound o-dianisidine. L-lysine alpha-oxidase catalyzed oxidative deamination of L-lysine with O2 consumption and formation of H2O2, NH3 and alpha-keto-epsilon-aminocaproic acid. Horseradish peroxidase and a non-carcinogenic compound 3,3,5,5'-tetramethylbenzidine dihydrochloride as an electron donor were used in determination of H2O2 formed. The procedure developed enabled also to measure the L-lysine alpha-oxidase activity at the enzyme concentrations of 10-500 ng/ml. The only limitation of the procedure is relatively low pH-values of the reaction medium.
Lukasheva
E V
EV
Institute of Applied Enzymology, Vilnius, Turku University, Finland.
Vesa
V S
VS
Korpela
T K
TK
Berezov
T T
TT
D000594:Amino Acid Oxidoreductases; D001560:Benzidines; D006735:Horseradish Peroxidase / Q000737:chemistry; D006861:Hydrogen Peroxide / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008239:Lysine / Q000032:analysis*; D013057:Spectrum Analysis; D014242:Trichoderma / Q000201:enzymology*
D001560:Benzidines; C021758:3,3',5,5'-tetramethylbenzidine; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase; D000594:Amino Acid Oxidoreductases; C041123:L-lysine oxidase; D008239:Lysine
Spektrofotometricheskiĭ metod opredeleniia kontsentratsii L-lizina s pomoshch'iu L-lizin-alpha-oksidazy iz Trichoderma sp. i 3,3',5,5'-tetrametilbenzidindigidrokhlorida.