响应面法优化酶解藻蓝蛋白制备抗肥胖肽

Optimization of Enayme Extraction of Anti-obesity Peptide from Phycocyanin by Response Surface Methodology

为了寻找具有减肥功效的天然产物,挖掘螺旋藻藻蓝蛋白的生物活性,以钝顶螺旋藻超声波诱变藻株40s Y为材料,水解度和胰脂肪酶抑制活性为指标,评价中性蛋白酶、碱性蛋白酶、木瓜蛋白酶、胰蛋白酶水解藻蓝蛋白的效果。选用效果最好的中性蛋白酶为试验用酶,在单因素试验基础上,采用响应面分析法建立藻蓝蛋白酶解条件的二次项数学模型,探讨酶解温度、酶解时间、加酶量、pH值对酶解效果的影响。结果表明,酶解时间、加酶量及两者交互作用的影响极显著,初始pH值及加酶量与酶解温度交互作用的影响显著。在酶解温度47℃,酶解时间4.5 h,加酶量118 U/m L,pH值7.0时,胰脂肪酶抑制率为53.88%,与响应面预测值54.96%基本吻合。此结果比单因素试验最大抑制率40.23%提高36.6%。

In order to find natural products with anti-obesity effect, bioactivity of phycocyanin was studied. The strain 40sY of Spirulina platensis, which is ultrasonic mutated, was used as the source material. The degree of hydrolysis and inhibitory activities of pancreas lipase were used as indicators to evaluate the effect of neutral proteinase, alkali pro- tease, papain and trypsin to phycocyanin hydrolysis. Neutral proteinase which has the most obvious effect was selected for further single-factor experiment. A quadratic-muhinominal model based on Response Surface Methodology （RSM）, was established to investigate the effects of temperature, time duration, neutral protease concentration and pH on enzy-matic hydrolysis. Results showed that the enzymatic hydrolysis time, amount of enzyme and their interaction have extremely significant effect, while initial pH value and interaction between enzyme amount and temperature have significant effect. Under the condition of temperature of 47℃, time duration of 4.5 h, neutral protease content of 118 U/mL and pH 7.0, the highest inhibitory rate of pancreas lipase is 53.88% which is closed to the forecast value of 54.96% by RSM. The present result increased by 36.6% compared to the highest inhibitory rate of 40.23% from a single-factor experiment.