河蚬多肽的制备工艺

以酸溶性多肽得率（TCA-SN）为指标,研究河蚬多肽的制备工艺.比较了中性蛋白酶Neutrase、复合蛋白酶Protamex、碱性蛋白酶Alcalase和木瓜蛋白酶Papain对河蚬蛋白的水解效果,考察了加酶量、p H、温度、时间、底物浓度五种因素对Alcalase碱性蛋白酶酶解过程的影响,采用正交实验优化法确定碱性蛋白酶Alcalase最佳酶解工艺条件为:酶解温度55℃,加酶量3%,底物浓度2%,起始p H8.5,酶解时间为3 h;在此条件下酸溶多肽的得率为29.65%。再利用大孔树脂吸附法对河蚬多肽进行分离,比较SQD-67、DA201-A、DA201-DⅡ、DA201-M和DA201-H几种树脂对河蚬多肽的吸附性能,最终确定大孔吸附树脂DA201-C的最佳吸附条件为:上样流速1 BV/h,上样浓度30 mg/m L,乙醇洗脱浓度为75%,洗脱液经减压浓缩、冷冻干燥得到河蚬多肽,其纯度为82.19%。 

Preparation technology of polypeptide from Corbicula fluminea

In this paper,with trichloroacetic acid- soluble nitrogen( TCA-SN) as indicators,the effect of several different enzymes including Neutrase,Protamex,Alcalase,Papain for protein hydrolysis were compared. Hydrolysis temperature,hydrolysis time,the dosage of enzyme,pH,the substrate concentration were as influencing factors. The results indicated that Alcalase was the best hydrolytic enzymes.The hydrolysis parameters was defined by the orthogonal test.The results showed that the optimum conditions were temperature of 55 ℃,the dosage of enzyme of 3%,the substrate protein concentration of 2%,the pH8.5,time of 3 h,and the TCA-SN was 26.95%. The adsorption capacity of several different macroporous adsorption resins( MAR) for Corbicula fluminea peptides were investigated. The DA201-C MAR had the optimum adsorption property for Corbicula fluminea peptides. The best condition was feeding amount of 1 BV/h,feeding concentration of 30 mg/mL,ethanol elution volume fraction of 75%,eluent were decompressed enrichment and freeze- drying,the percentage composition of peptides was 82.19%.