| 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4.5 h，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。 |
| 关键词: 牛乳酪蛋白酶解物 乳基料 分离纯化 抗菌肽 |
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| 基金项目: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） |
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| Isolation,purification of bovine casein hydrolyzates and preparation of antimicrobial peptide dairy ingredients |
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| Abstract: |
| 【Objective】Bovine casein hydrolyzates were isolated and purified,and their antimicrobial activities were detected.Antimicrobial peptide dairy ingredients were prepared as well.【Method】A protease was chosen from neutral protease,alcalase,trypsin and papain to hydrolyze bovine casein,and its hydrolyzates were isolated and purified by macroporous resin and gel filtration chromatography.Diameter of antimicrobial circle,MIC and MBC against Escherichia coli,Salmonella,and Staphylococcus aureuswere 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 bovine 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 antibacterial 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 absorption resin and gel filtration chromatography.The bovine casein hydrolyzates could be used to prepare antimicrobial peptide dairy ingredients. |
| Key words: bovine casein hydrolyzates dairy ingredients isolation and purification antimicrobial peptide |
