蛋白酶-Sevag法去除木瓜总多糖中蛋白质的工艺优选

Optimization of Protein Removal Technology for Total Polysaccharides in Chaenomelis Fructus by Papain Enzyme-Sevag Method

目的:优选蛋白酶-Sevag法去除木瓜总多糖中蛋白质的工艺条件。方法:以总多糖保留率和蛋白脱除率为评价指标,通过单因素试验考察酶解pH和Sevag法处理次数对去除工艺的影响,利用星点设计-效应面法考察酶底质量分数、酶解时间及温度对酶解工艺的影响。采用UV测定蛋白质含量,检测波长280 nm。结果:最佳工艺条件为酶解pH 7,时间89 min,温度54℃,酶底质量分数1.70%,Sevag法处理数1次;总多糖保留率70.04%,蛋白脱除率63.42%,与预测值的偏差分别为1.42%和0.17%。结论:优选的工艺条件预测性良好,在有效去除蛋白的同时可尽可能地避免总多糖的损失,减少有机溶剂的使用次数。

Objective：To optimize technology conditions of protein removal from total polysaccharides in Chaenomelis Fructus by papain enzyme-Sevag method.Method：Taking ratios of total polysaccharides retention and proteins removal as indexes,central composite design-response surface methodology was used to optimize technology conditions of enzymatic hydrolysis with enzyme concentration,hydrolysis time and temperature as independent variables,single factor experiments were adopted to optimize pH and number of Sevag.The content of proteins was determined by UV,detection wavelength was set at 280 nm.Result：Optimal technology conditions were hydrolysis pH 7,the mass fraction of enzyme substrates 1.70％,hydrolysis temperature at 54 ℃,hydrolysis time 89 min,treatment once of Sevag.Under these conditions,ratios of total polysaccharides retention and proteins removal were 70.04％ and 63.42％,whose deviations were 1.42％ and 0.17％ by comparing with their predicted values,respectively.Conclusion：This optimized had highly predictive,it could effectively remove proteins,at the same time,avoid loss of total polysaccharides as much as possible,reduce use of organic solvents.