基于响应面法的花豇豆酯酶提取工艺

Extraction Technology of Cowpea Esterase Based on Response Surface Methodology

基于酶抑制原理建立果蔬中农药残留的快速检测方法,以固蓝B盐作为显色剂,通过比较面粉、麦麸和13种植物种子的总酯酶活力和比活力,确定用于农药残留检测的最佳植物酯酶源。结果表明,花豇豆酯酶的总酯酶活力和比活力最大,确定花豇豆为最佳植物酶源。结合单因素试验、响应面试验和验证试验确定植物酯酶最佳提取条件:料液比1︰5(g/mL)、提取液pH 8、浸提时间2 h、离心速率8000 r/min,此最佳提取条件下酶活力为9.081 U/m L。因此,在该条件下能够较好地从花豇豆中提取植物酯酶,并且能够保持较高的酶活力。

Based on the principle of enzyme inhibition,a rapid detection method for pesticide residues in fruits and vegetables was established.Using fast blue B salt as a color developing agent,by comparing the total esterase activity and specific activity of flour,wheat bran and 13 plant seeds,the best plant esterase source for pesticide residue detection was determined.The results showed that the total esterase activity and specific activity of cowpea esterase were the largest,and cowpea was determined as the best plant enzyme source.Combined with single factor experiment,response surface experiment and verification experiment,the optimum extraction conditions of plant esterase were determined as follows:the ratio of material to liquid was 1︰5(g/mL),the pH of the extract was 8,the extraction time was 2 h,and the centrifugal rate was 8000 r/min.Under this optimum extraction condition,the enzyme activity was 9.081 U/mL.Therefore,under these conditions,plant esterases could be better extracted from cowpea,and high enzyme activity could be maintained.