酮氨氧化酶（KAOD）

用途 用于酶法糖化白蛋白试剂的研发和大量配制。 酶学性质（注意-4、 2+为上标） 来源： 微生物 酶学委员会编号： EC1.5.3 分子量： 55kDa (SDS-PAGE) 等电点： 6 Km值： 5.0×10⁻⁴ M (Fructosyl-Ala) 抑制剂： Hg²⁺ , Pb²⁺ 最适pH： 7.7 图1 最适温度： 42℃ 图2 pH稳定性： 5.0-9.5 (25℃,16h) 图3 热稳定性： 40℃以下稳定(pH8.0，30min) 图4 稳定性： -25～-15℃静置保存12个月保持 90%以上活性 图5 保护剂： 甘油、海藻糖 1、原理 2、酶活定义 单位酶活定义为在下述条件下每分钟催化生成1μmol H2O2所需的酶量。 3、试剂准备 试剂I：1M磷酸钾缓冲液，pH 8.0 试剂II：1kU/mL POD 试剂III：50mM TOOS溶液 试剂IV：50mM 4-AA溶液 试剂V：200mM 糖化丙氨酸 酶稀释液：20mMTris-HCl, pH 8.0 按如下配制反应混合物： 试剂I：10mL 试剂II：0.1mL 试剂III：1mL 试剂IV：1mL 试剂V：10mL 双蒸水：定容至100mL 4、操作步骤 4.1 向1mL比色皿中加入980μL反应混合物。 4.2 37°C孵育5分钟。 4.3 向反应混合物中加入20μL待测酶液。 4.4 37°C反应，用分光光度计在555nm检测样品1min内的吸光度变化（∆As）。 *空白对照测定方法：用20μL酶稀释液代替待测酶液，测定样品1min内的吸光度变化（∆A b ）。 ∆A=∆As-∆ b 5、 活力计算 Vt：反应液总体积(1.0mL)； Vs：酶液体积(0.02mL)； t：反应时间(1min)； df：稀释倍数； C：酶浓度(mg/mL)； 1.0：光路长度 (cm)； 1/2：1 摩尔过氧化氢生成1/2摩尔醌亚胺染 料； 39.2：标准反应条件下，生色基团在555nm 处毫摩尔吸光系数(cm²⁺ /μmol)。 Application It is used in the research and development of enzymatic glycosylated albumin reagents and mass formulation. Enzymatic properties Source: Microorganism Enzymology Committee Number: EC1.5.3 Molecular weight: 55kDa (SDS-PAGE) Isoelectric point: 6 Km value: 5.0×10 ⁻⁴ M (Fructosyl-Ala) Inhibitors: Hg ²⁺ , Pb ²⁺ Optimal pH: 7.7 Figure 1 Optimum temperature: 42℃ Figure 2 pH stability: 5.0-9.5 (25℃, 16h) Figure 3 Thermal stability: Stable below 40℃ (pH8.0, 30min) Figure 4 Stability: -25 ~ -15℃ standing store for 12 months More than 90% activity Figure 5 Protective agent: glycerin, trehalose Assay method for activity 1. Principle 2. Definition of enzyme activityUnit enzyme activity is defined as the amount of enzyme required to catalyze the production of 1μmol H2O2 per minute under the following conditions. 3. Reagent preparation Reagent I: 1M potassium phosphate buffer, pH8.0 Reagent II: 1kU/mL POD Reagent III: 50mM TOOS solution Reagent IV: 50mM 4-AA solution Reagent V: 200mM Glycated alanine Enzyme diluent: 20mM Tris-HCl, pH8.0 Prepare the reaction mixture as follows: Reagent I: 10mL Reagent II: 0.1mL Reagent III: 1mL Reagent IV: 1mL Reagent V: 10mL Double steam water set volume to 100mL 4. Operation procedure 4.1 Add 980μL reaction mixture into 1mL colorimetric dish. 4.2 Incubate at 37°C for 5 minutes. 4.3 Add 20 μL of enzyme solution to be tested to the reaction mixture. 4.4 Reaction at 37°C, the absorbance change (∆As) of the sample within 1min is detected by spectrophotometer at 555nm. * Blank control measurement method: Use 20 μL of enzyme dilution solution instead of the enzyme solution to be tested, and measure the absorbance change (ΔA b ) of the sample within 1 minute. ∆A=∆As-∆A b 5. Vitality computing Vt: Total volume of reaction liquid (1.0 mL); Vs: Enzyme liquid volume (0.02 mL); t: Reaction time (1 min); df: Dilution ratio; C: Enzyme concentration (mg/mL); 1.0: Optical path length (cm); 1/2: 1 mole hydrogen peroxide to generate 1/2 mole quinone imide dye; 39.2: Under standard reaction conditions, the millimolar absorption coefficient of the color group at 555nm (cm² /μmol).

No numbering system information yet

No physical and chemical property information yet

No safety information yet

No production method and use information yet

No upstream information yet

No downstream information yet

No related product information yet