小麦酯酶的提取及反应条件的研究

Study on extraction and reaction conditions of wheat esterase

以小麦粉为原料提取植物酯酶,根据植物酯酶催化α-乙酸萘酯和固蓝B盐反应生成紫红色偶氮化合物,再采用紫外分光光度计在波长为520 nm时测定其吸光度值,确定最佳提取及反应条件.结果表明:小麦酯酶提取的最佳条件为浸泡时间24 h,离心转数4 000 r/min,离心时间10 min,料液比1∶4;小麦酯酶催化α-乙酸萘酯和固蓝B盐的最佳反应条件为显色温度35℃、显色时间20 min、p H=5.5、显色基质加入量200μL、酶液加入量150μL,且各因素的主次顺序为酶液加入量极差最大,其次分别为显色温度、显色时间、p H值.

The plant esterase was extracted from wheat flour as raw material. According to the principle that the plant esterase catalyzed the reaction of α-naphthyl acetate and solid blue B salt to produce purple azo compound, the UV spectrophotometer was used at the wavelength of 520 nm to measure absorbance values to determine the best extraction and reaction conditions.The results showed that the optimal conditions for the extraction of wheat esterase were as follows ： immersion time 24 h, centrifugal rotation 4 000 r/min, centrifugation time 10 min , ratio 1 ： 4.The optimum reaction conditions were as follows：color temperature 35 ℃ , color developing time 20min , pH = 5.5, coloring substrate addition amount 200 μL, enzyme dosage 150μL, and the primary and secondary order of the factors is the maximum liquid volume, followed by the color temperature , color development time,pH value.