双酶法制备马鹿茸降血糖肽工艺优化及其对α-葡萄糖苷酶的抑制效果  

Optimization of Preparation of Hypoglycemic Peptides from Red Deer(Cervus elaphus) Antlers by Two-Step Enzymatic Hydrolysis and Their α-Glucosidase Inhibitory Activity

作　　者：包美丽[1,2] 杨添植 张立钢[3] 赵玉红[1] BAO Meili;YANG Tianzhi;ZHANG Ligang;ZHAO Yuhong(School of Forestry, Northeast Forestry University, Harbin 150040, China;Heilongjiang Vocational College for Nationalities, Harbin 150066, China;College of Food Science, Northeast Agricultural University, Harbin 150030, China)

机构地区：[1]东北林业大学林学院,黑龙江哈尔滨150040 [2]黑龙江民族职业学院,黑龙江哈尔滨150066 [3]东北农业大学食品学院,黑龙江哈尔滨150030

出　　处：《食品科学》2017年第6期88-95,共8页Food Science

基　　金：中央高校基本科研业务费专项(2572014CAQ02);黑龙江省高校科技成果产业化前期研发培育项目(1254CGZH14)

摘　　要：为探索制备马鹿茸降血糖肽的最佳工艺条件,以α-葡萄糖苷酶抑制率为指标,从碱性蛋白酶、风味蛋白酶、中性蛋白酶和胰蛋白酶中筛选出两种酶,根据其体外降血糖效果确定酶的作用顺序,再以水解度、α-葡萄糖苷酶抑制率和蛋白质回收率为指标进行单因素试验和正交试验,优化降血糖肽制备工艺条件。结果表明:碱性蛋白酶和风味蛋白酶比中性蛋白酶和胰蛋白酶更适合用于制备马鹿茸降血糖肽。采用碱性蛋白酶-风味蛋白酶顺序对马鹿茸进行水解,所得酶解产物的α-葡萄糖苷酶抑制率、蛋白质回收率和水解度较高,分别为21.11%、39.12%、19.88%。通过单因素试验和正交试验确定双酶酶解最佳工艺条件为先用碱性蛋白酶在p H 8.0、60℃、底物质量分数12%、加酶量5 000 U/g条件下酶解3 h,再用风味蛋白酶于p H 6.5、45℃、底物质量分数5%、加酶量6 000 U/g条件下酶解1 h。双酶分步水解终产物的α-葡萄糖苷酶抑制率受质量浓度的影响,当质量浓度为3 mg/m L时,α-葡萄糖苷酶抑制率可达94.09%,IC50值为1.82 mg/m L。碱性蛋白酶-风味蛋白酶双酶分步水解马鹿茸可获得高α-葡萄糖苷酶抑制率的降血糖肽。This study was designed to optimize the enzymatic preparation of hypoglycemic peptides from red deer(Cervuselaphus)antlers.Alcalase,flavourzyme,neutral protease and trypsin were comparatively evaluated based onα-glucosidaseinhibitory activity of hydrolysates to select two suitable enzymes for sequential hydrolysis of red deer antlers.The sequencesof two hydrolysis steps were compared for better hypoglycemic activity in vitro and the hydrolysis conditions wereoptimized based on degree of hydrolysis(DH),α-glucosidase inhibitory activity and protein recovery with one-factor-at-atimemethod and orthogonal array design.The results showed that alcalase and flavourzyme were found to be more suitableand sequential use of alcalase followed by flavourzyme was optimal for higher percentage inhibition ofα-glucosidaseactivity,DH and protein recovery,reaching21.11%,19.88%and39.12%,respectively.The optimal hydrolysis conditionswere obtained as follows.Red deer antlers were hydrolyzed firstly with5000U/g alcalase at an initial pH of8.0and60℃with a substrate concentration of12%for3h,and then with6000U/g flavourzyme for1h at45℃with a substrateconcentration of5%after pH adjustment to6.5.The final hydrolysate exhibited concentration-dependentα-glucosidaseinhibitory activity,with a percentage inhibition of94.09%at3mg/mL and IC50value of1.82mg/mL.It was concluded thatalcalase-flavourzyme hydrolysis would provide an efficient way to prepare hypoglycemic peptides from red deer antlers.

关 键 词：马鹿茸 酶解 降血糖肽 

分 类 号：O629.72[理学—有机化学]