Voprosy meditsinskoi khimii (ISSN 0042-8809)

Spectrophotometric micromethod to determine L-lysine-alpha-oxidase activity

   
Smirnova I.P., Siatkin S.P., Berezov T.T.
PubMed Id: 6538725
Year: 1984 vol: 30  issue:1  pages: 133-136
Abstract: A spectrophotometric micromethod is developed for estimation of L-lysine-alpha-oxidase. The method is based on measurement of hydrogen peroxide formed in enzymatic oxidation of L-lysine. Optimal conditions were developed for estimation of the enzymatic activity in the extracts of Trichoderma sp. The saturating concentration of L-lysine was 10 mM. Km values for L- and DL-lysines constituted 3.0 . 10(-3) M and 6.4 . 10(-4) M, respectively. The reaction proceeded without a latent phase and H2O2 formation versus time plot had a linear shape within 20 min. The enzyme optimal activity was found at pH 5.8-6.0. The best buffer, required for lysine oxidation in the reaction, proved to be phosphate, but not succinate, borate or glycine buffers. The method described was highly sensitive and reproducible.
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Reference: Smirnova I.P., Siatkin S.P., Berezov T.T., Spectrophotometric micromethod to determine L-lysine-alpha-oxidase activity, Voprosy meditsinskoi khimii, 1984, vol: 30(1), 133-136.
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