糖基化亚硝基血红蛋白制备工艺优化

摘要

以猪血红蛋白为原料制备亚硝基血红蛋白,根据热稳定性选择用壳聚糖对亚硝基血红蛋白进行糖基化。对糖基化反应过程中亚硝基血红蛋白与壳聚糖的质量分数比例、pH值、加热温度和时间对反应产物浓度（吸光度）的影响进行研究。单因素试验及响应面试验结果表明,糖基化亚硝基血红蛋白制备的最佳工艺条件为亚硝基血红蛋白与壳聚糖的质量分数比例为4.16：1、pH5.6、加热温度50.5℃、时间15.7min。%Nitrosohemoglobin was prepared from porcine hemoglobin and glycosylated with chitosan,which was selected in consideration of thermal stability.The effects of mass ratio of nitrosohemoglobin to chitosan,pH,temperature and reaction time on the glycosylation of nitrosohemoglobin were probed by one-factor-at-a-time method.Subsequently,response surface methodology,based on Box-Benhnken experimental design,was used to optimize pH,temperature and reaction time for achieving maximum absorbance at 450 nm of reaction products.The optimal glycosylation conditions of nitrosohemoglobin were determined as follows： mass ratio of nitrosohemoglobin to chitosan,4.16：1,pH 5.6,temperature 50.5 ℃ and 15.7 min.