胰蛋白酶对猪血红蛋白酶解脱色条件优化  

Optimization of Conditions for Decolorization of Porcine Hemoglobin by Trypsin

作　　者：姚伦欢 郝刚[1] 唐善虎[1] 李思宁[1] YAO Lunhuan;HAO Gang;TANG Shanhu;LI Sining(College of Food Science and Technology,Southwest University for Nationalities,Chengdu 610041,China)

机构地区：[1]西南民族大学食品科学与技术学院,四川成都610041

出　　处：《肉类研究》2020年第10期19-25,共7页Meat Research

基　　金：“十三五”国家重点研发计划重点专项(2018YFD0400101);西南民族大学研究生创新项目(CX2019SZ160)。

摘　　要：以酶解液水解度(hydrolysis degree,DH)、三氯乙酸(trichloroacetic acid,TCA)可溶性氮含量及血红素残留量为评定指标,采用单因素试验比较胰蛋白酶和碱性蛋白酶对猪血红蛋白的酶解脱色效果。在单因素试验基础上,进行4因素3水平正交试验。结果表明:胰蛋白酶水解脱色的效果更好,最佳酶解工艺参数为酶底比7500 U/g、酶解温度45℃、pH 7.5、底物质量浓度8 g/100 mL,在此条件下,酶解液DH为25.67%,TCA可溶性氮含量为80.05%,血红素残留量为18.34%,猪血红蛋白多肽粉蛋白含量83.17%;对此条件下制备的猪血红蛋白多肽粉进行分子质量分布分析,分子质量在1000 Da以下的小肽占比94.20%;最佳酶解脱色条件下,猪血红蛋白多肽粉中TCA可溶性氮含量为80.05%的蛋白占猪血红蛋白多肽粉蛋白含量的96.25%,这与分子质量分布结果比较符合。This study comparatively evaluated the decolorization efficiency of porcine hemoglobin with trypsin and alkaline protease based on hydrolysis degree(DH),trichloroacetic acid(TCA)soluble nitrogen content and heme residue.Trypsin was determined to be more suitable for decolorizing porcine hemoglobin than alkaline protease.Using a fourfactor,three-level orthogonal array design,the optimal hydrolysis conditions for decolorizing porcine hemoglobin by trypsin were determined as follows:enzyme/substrate ratio 7500 U/g,temperature 45℃,pH 7.5,and substrate concentration 8 g/100 mL.Under these conditions,the DH was 25.67%,and the TCA soluble nitrogen content,heme residue and total protein content of the peptide obtained powder were 80.05%,18.34%and 83.17%,respectively.Molecular mass distribution analysis showed that small peptides with molecular masses less than 1000 Da accounted for 94.20%of the peptide powder.TCA soluble nitrogen accounted for 96.25%of the protein content in the peptide powder,which is quite consistent with the results of molecular mass distribution.

关 键 词：猪血红蛋白 酶解 水解度 三氯乙酸可溶性氮 血红素 

分 类 号：TS251.9[轻工技术与工程—农产品加工及贮藏工程]