摘要
为了明确芝麻脂肪氧化酶活性测定的适宜条件,采用紫外分光光度法测定芝麻脂肪氧化酶的活性并对测定的适宜条件进行探讨。结果表明,芝麻脂肪氧化酶活性测定的最佳反应条件为:取2.95 m L 0.05 mol/L的磷酸缓冲液(p H值6.2)、20μL 10 mmol/L的底物亚油酸钠、50μL酶液迅速混匀后,立即在紫外光234 nm处观察OD值的变化,最适检测反应时间为0~1.5 min。将吸光度代入公式计算酶活性。此外,芝麻脂肪氧化酶活性随酶液保存时间的延长而下降,应将酶液保存在4℃条件下并于1 h内进行测定。酶液添加量为10~50μL时,酶活力(y)与酶液添加量(x)成正比,y=0.084 6x+5.858,且线性关系良好(R2=0.999 8)。精密度试验结果显示RSD为0.027%,建立的方法方便快捷,稳定性好,可广泛使用。
In order to ascertain the suitable conditions for the determination of lipoxygenase activity in sesame,the activity of lipoxygenase in sesame was determined by ultraviolet spectrophotometry and the measured conditions were discussed. The results showed that the optimal reaction conditions were as follows : 2.95 mL of phosphate buffer (0.05 mol/L, pH 6.2) ,20μL of sodium linoleate ( 10 mmol/L) as the substrate,and 50μL of enzyme solution. Blend them quickly and immediately observe the change of OD value at 234 nm UV light. The optimal reaction time was 0-1.5 min. The enzyme activity was calculated using the equation according to the absorbance value. In addition,the activity of lipoxygenase in sesame decreased with the elongation of the storage time of the enzyme solution, which should be kept at 4℃ and measured within 1 h. When the added volume of the enzyme solution was between 10-50μL,the enzymatic activity (y) was in direct proportion to the enzyme volume (x) ,y = 0. 084 6x + 5. 858, and the linear relationship was good (R^2 = 0. 999 8). The results of the precision test showed that the RSD was 0. 027% ,and the established method was convenient,stable and suitable for wide application.
出处
《河南农业科学》
CSCD
北大核心
2017年第11期48-51,共4页
Journal of Henan Agricultural Sciences
基金
公益性行业(农业)科研专项(201303072)