富血红素多肽产物制备用酶的比较研究

Comparison of Proteases for Hydrolyzing Porcine Hemoglobin

为筛选适于生产富血红素多肽产物的蛋白酶,比较不同蛋白酶对猪血红蛋白的水解效果。在AS1398中性蛋白酶、2709碱性蛋白酶、木瓜蛋白酶和胰蛋白酶的最适水解pH值及温度下,以L16(45)正交试验研究酶种类、酶质量分数、底物质量分数和水解时间等因素对水解率、水解液中粗多肽含量和血红素含量等指标的影响。结果表明:酶种类和水解时间两因素对水解率指标的影响达到极显著水平(P<0.01);底物质量分数是影响粗多肽含量指标的最重要因素;酶种类对血红素含量指标的影响则达到显著性水平(P<0.05)。中性蛋白酶有利于提高水解率,木瓜蛋白酶有利于提高水解物的粗多肽含量,而碱性蛋白酶则有利于提高水解物的血红素含量。

In order to screen a protease suitable for producing heme-enriched polypeptides, the effects of various proteases on hydrolysis efficiency of porcine hemoglobin were compared. Under the optimal pH and temperature of proteases such as AS1398 neutral protease, 2709 alkaline protease, papain and trypsin, orthogonal experiments were used to explore the effects of enzyme type, enzyme concentration, substrate concentration and hydrolysis time on the hydrolysis degree of porcine hemoglobin and the contents of polypeptides and heme. Results showed that the effects of enzyme type and hydrolysis time on the degree of hydrolysis reached an extremely significant level （P〈0.01）. Substrate concentration was the most important factor to affect the content of polypeptide production. The effect of enzyme type on heme content reached a significant level （P〈0.05）. Neutral protease is beneficial for the increase of hydrolysis degree and papain is good for the increase of polypeptide content, while alkali protease is helpful for the increase of heme content.