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分别采用体积排阻色谱和Folin-酚法测定了经酸性蛋白酶(蛋白酶M、胃蛋白酶)、中性蛋白酶(蛋白酶A、蛋白酶N)、枯草杆菌碱性蛋白酶Alcalase酶解后制得的大豆肽经过模拟体内消化之后分子量的变化,以及离子强度对酶解物聚合的影响。大豆肽的高效液相色谱体积排阻色谱图显示,50%左右的大豆肽的分子量集中在10~30 kDa之间;在经过模拟肠道胃蛋白酶和胰蛋白酶消化之后,分子量范围在3~30 kDa的肽段变化相对较大。Folin-酚法测定酶解上清液中肽的浓度结果表明,胃-肽和A-肽酶解之后添加NaCl能明显减弱聚合现象,N-肽酶解之前添加NaCl能明显减弱聚合现象,而不管哪种方式添加NaCl M-肽都会加剧聚合现象,Alcalase-肽的浓度始终维持在11.40 mg·mL-1左右,离子强度对其影响不大。
Soybean peptides were obtained by enzymolysis of Alcalase with acid protease (protease M, pepsin), neutral protease (protease A, protease N) and Bacillus subtilis alkaline protease using size exclusion chromatography and Folin-phenol method respectively The change of molecular weight after in vivo digestion was simulated, and the effect of ionic strength on the enzymolysis polymerization was simulated. High performance liquid chromatography with size exclusion chromatography of soy peptides showed that about 50% of the soy peptides concentrated in the molecular weight of 10 ~ 30 kDa; after simulated intestinal pepsin and trypsin digestion, the molecular weight range of 3 ~ 30 kDa peptide changes relatively large. Folin-phenol method to determine the concentration of peptides in the enzymolysis supernatant The results showed that the addition of NaCl after gastric-peptide and A-peptide enzymatic hydrolysis significantly reduced the polymerization phenomenon. The addition of NaCl prior to N-peptide digestion significantly reduced the polymerization, The addition of NaCl M-peptide in various ways would aggravate the polymerization phenomenon. The concentration of Alcalase-peptide remained at about 11.40 mg · mL-1, but the ionic strength had little effect on it.