牛乳酪蛋白酶解物的分离纯化及抗菌肽乳基料的制备研究  

Isolation,purification of bovine casein hydrolyzates and preparation of antimicrobial peptide dairy ingredients

作　　者：熊清权[1] 李志成[1] 童伟[1] 郑晓莹[1] 苏晋文[1] 葛武鹏[1] 任娇[1] 

机构地区：[1]西北农林科技大学食品科学与工程学院,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2013年第1期200-206,共7页Journal of Northwest A&F University(Natural Science Edition)

基　　金：陕西省农业攻关项目(2011K01-04);陕西省科技统筹创新工程计划项目(2011KTCL02-11);国家"十二五"科技支撑计划项目(2012BAD12B07)

摘　　要：【目的】分离纯化牛乳酪蛋白酶解物,检测酶解物及其分离组分的抑菌活性,并制备抗菌肽乳基料。【方法】从中性蛋白酶、碱性蛋白酶、胰蛋白酶及木瓜蛋白酶中筛选一种蛋白酶水解牛乳酪蛋白,将其酶解产物经大孔吸附树脂和凝胶过滤色谱分离、纯化后,以对大肠杆菌、沙门氏菌和金黄色葡萄球菌的抑菌圈直径、最小抑菌浓度(MIC)、最小杀菌浓度(MBC)为指标,分析酶解物和分离组分的抑菌活性。【结果】(1)木瓜蛋白酶水解牛乳酪蛋白4.5h,其酶解物对大肠杆菌、沙门氏菌和金黄色葡萄球菌均有较好的抑制作用。(2)大孔吸附树脂将牛乳酪蛋白酶解物分离成4个组分,其中以体积分数75%乙醇洗脱组分的抑菌作用较强。(3)凝胶过滤色谱将体积分数75%乙醇洗脱组分分离成4个色谱峰,其中A峰对大肠杆菌和沙门氏菌抑制作用较强,B峰对金黄色葡萄球菌抑制作用较强,A峰和B峰经冷冻干燥后成功制备了2种抗菌肽乳基料。【结论】牛乳酪蛋白酶解物经大孔吸附树脂和凝胶过滤色谱分离、纯化后,其分离组分的抑菌活性逐步增强,能够用于抗菌肽乳基料的制备。[Objective] Bovine casein hydrolyzates were isolated and purified, and their antimicrobial ac- tivities were detected. Antimicrobial peptide dairy ingredients were prepared as well. [Method] A protease was chosen from neutral protease,alealase,trypsin and papain to hydrolyze bovine casein, and its hydroly- zates were isolated and purified by macroporous resin and gel filtration chromatography. Diameter of anti- microbial circle,MIC and MBC against Escherichia coli, Salmonella, and Staphylococcus aureus were used to analyzed the antimicrobial activities of hydrolyzates and their separated components. [Result] （1） Bovine casein was hydrolyzed for 4.5 h by papain, and its hydrolyzates had stronger antimicrobial effects against Escherichia coli ,Salmonella,and Staphylococcus aureus. （2） Macroporous resin was used to separate bo- vine casein hydrolyzates into 4 groups,and group eluted by ethanol with a volume ratio of 75 % showed the strongest antibacterial effects. （3） The group of hydrolyzing components eluted by ethanol with a volume ratio of 75% were further isolated to 4 fractions by gel filtration chromatography. Fraction A showed strong antibacterial activity against Escherichia coli and Salmonella,and fraction B showed strong antibac- terial activity against Staphylococcus aureus. Two dairy ingredients of antimicrobial peptide from fractions A and B had been prepared successfully by lyophilization. [Conclusion] Antimicrobial activities of fractions from bovine casein hydrolyzates increased gradually after isolation and purification by macroporous absorp- tion resin and gel filtration chromatography. The bovine casein hydrolyzates could be used to prepare anti- microbial peptide dairy ingredients.

关 键 词：牛乳酪蛋白酶解物 乳基料 分离纯化 抗菌肽 

分 类 号：Q814.9[生物学—生物工程] TQ464.7[化学工程—制药化工]