正交试验优化哈蟆油蛋白双酶法水解工艺  

Optimization of Two-Step Enzymatic Hydrolysis of Oviductus Ranae Protein by Orthogonal Array Design

作　　者：董颖[1] 张鹤[1] 周光新[1] 赵大庆[1] 赵雨[1] 丛德毓[1] 

机构地区：[1]长春中医药大学,吉林长春130117

出　　处：《食品科学》2014年第6期36-39,共4页Food Science

基　　金："重大新药创制"重大科技专项(2011ZX09401-305-09);吉林省科技发展计划项目(YYZX201126)

摘　　要：目的:考察双酶法制备哈蟆油蛋白的最佳工艺。方法:以哈蟆油为原料,采用柠檬酸浸提和水煎煮的方法提取哈蟆油蛋白,先后加入胃蛋白酶和碱性蛋白酶进行酶解,根据酶解后哈蟆油蛋白相对分子质量分布,应用单因素和正交试验确定哈蟆油蛋白酶解的最佳条件。结果:最佳酶解条件为胃蛋白酶与底物比(m/m)1∶2 000、酶解时间2 h、碱性蛋白酶与底物比(m/m)1∶100、酶解时间6 h,酶解后哈蟆油蛋白相对分子质量小于5 000所占的比例为94.85%,哈蟆油蛋白中蛋白质含量为49.85%。结论:胃蛋白酶与碱性蛋白酶结合使用可以有效地降低哈蟆油蛋白的相对分子质量,利于人体吸收。Objective： To optimize the two-step enzymatic hydrolysis of Oviductus Ranae protein. Methods： Oviductus Ranae protein was extracted by soaking in citric acid, homogenizing Oviductus Ranae and then decoting the homogenate with water, and sequentially hydrolyzed with pepsin and alkaline protease. Various hydrolysis parameters were optimized by single factor and orthogonal array design methods to obtain maximum percentage of peptides with molecular weight less than 5 000. Results： The optimum hydrolysis conditions were obtained as follows： pepsin hydrolysis at an enzyme/substrate ratio of 1：2 000 （m/m） for 2 h followed by alkaline protease hydrolysis at an enzyme/substrate ratio of 1：100 （m/m） for 6 h. The percentage of peptides with molecular weight less than 5 000 in the hydrolysate obtained under these conditions was as high as 94.85%, and the protein content of the hydrolysate was 49.85%. Conclusion： The combined use of pepsin and alkaline protease can effectively decompose Oviductus Ranae protein for better absorption by the body.

关 键 词：哈蟆油蛋白 胃蛋白酶 碱性蛋白酶 双酶法 

分 类 号：R284.2[医药卫生—中药学]