乳铁蛋白水解物修饰物的分离纯化及其抗大肠杆菌活性  

Isolation and purification of lactoferrinhydrolysate's modifier and determination of its antibacterial activity on Escherichia coli

作　　者：马春敏 薄力影[2] 范婧[2] 李铁晶 

机构地区：[1]辽宁大学,辽宁大学轻型产业学院,辽宁沈阳110000 [2]乳品科学教育部重点实验室,东北农业大学,黑龙江哈尔滨150030

出　　处：《食品工业科技》2018年第1期92-95,106,共5页Science and Technology of Food Industry

基　　金：国家自然基金(31201337).

摘　　要：利用胃蛋白酶水解乳铁蛋白得到最小抑菌浓度为200 mg/m L的乳铁蛋白水解物(LHS),进行类蛋白反应导入色氨酸(Trp)得到乳铁蛋白水解物修饰物。对LHS及其色氨酸修饰产物采用葡聚糖凝胶过滤色谱G-25进行分离纯化,收集出峰产物,比较各级产物的抗大肠杆菌活性。结果表明,LHS和Trp修饰物分离纯化后的峰1为抑菌活性峰,各级产物的抑菌效果为LHS<LHS纯化峰1产物<Trp修饰产物<Trp修饰产物纯化峰1产物。加样浓度200 mg/m L时,LHS、Trp修饰物对大肠杆菌的抑菌率分别为40.42%和85.83%。加样浓度100 mg/m L时,LHS纯化峰1产物、Trp修饰物、Trp修饰物纯化峰1产物对大肠杆菌抑菌率分别为41.12%、58.31%和88.00%。可见,色氨酸修饰物对大肠杆菌的抑菌率较未修饰前提高了45.41%,色氨酸修饰产物经过纯化后对大肠杆菌的抑菌率较未纯化前提高了29.69%。将收集的活性峰作为样品,采用反相高效液相色谱进一步分离鉴定,证明经过分离纯化得到了纯度相对较高且活性提高的抗菌肽。The lactoferrinhydrolysate（LHS） with a minimum inhibitory bacterial concentration of 200 mg/mL was hydrolyzed by pepsin.And then the lactoferrinhydrolysate was modified with tryptophan（Trp） LHS and its tryptophan modifier were separated and purified by Sephadex G-25.The peak products were collected and their antibacterial activity was determined.Especially, the purred peak 1 belong to the antibacterial activity peak and the antibacterial effect of the separative products were LHS 〈 LHS＇ s peak 1 product 〈 Trp modifier 〈 Trp modifier＇ s peak 1 product.The inhibitory rates of LHS and Trp modifier on Eseheriehia coli were 40.42% and 85.83% at the concentration of 200 mg/mL. When adding the concentration of 100 mg/mL, the inhibitory rates of LHS＇ s peak 1 product,Trp modifier and Trp modifier＇ s peak 1 product on Escheriehia coli were 41.12% ,58.31% and 88.00% ,respectively. Then, the inhibition rate of tryptophan modifier against Escherichia coli was 45.41% higher than its unmodifier.The inhibition rate of tryptophan modifier＇ s purified product against Escherichia coli was 29.69% higher than its not purified product. The collected products were further separated and identified by reversed- phase high performance liquid chromatography（RP-HPLC） .The result demonstrated that the antibacterial peptide with high purity and high activity was obtained by separation and purification.

关 键 词：乳铁蛋白水解物修饰物 类蛋白反应 分离纯化 抑菌活性 反相高效液相色谱 

分 类 号：TS201.3[轻工技术与工程—食品科学]