Smirnova I P, Siatkin S P, Berezov T T
Vopr Med Khim. 1984 Jan-Feb;30(1):133-6.
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.
开发了一种用于测定L-赖氨酸-α-氧化酶的分光光度微量法。该方法基于对L-赖氨酸酶促氧化过程中形成的过氧化氢的测量。确定了用于测定木霉属提取物中酶活性的最佳条件。L-赖氨酸的饱和浓度为10 mM。L-赖氨酸和DL-赖氨酸的Km值分别为3.0×10⁻³ M和6.4×10⁻⁴ M。反应无潜伏期，过氧化氢生成量与时间的关系图在20分钟内呈线性。酶的最佳活性在pH 5.8 - 6.0时发现。反应中赖氨酸氧化所需的最佳缓冲液是磷酸盐缓冲液，而不是琥珀酸盐、硼酸盐或甘氨酸缓冲液。所描述的方法具有高度敏感性和可重复性。
