摘要
以鸽血红蛋白为原材料,采用在线数据库ExPASy Peptide Cutter中的蛋白酶模拟酶解,通过生物信息学工具筛选多肽潜在的生物活性、溶解性、毒性和致敏性,由此预测最适宜制备鸽血红蛋白源抗氧化肽的蛋白酶。设置不同的酶解时间、酶添加量和酶解温度作为单因素,探究不同酶解条件对酶解产物水解度和DPPH自由基清除率的影响。在单因素试验的基础上,通过响应面法优化试验,探究酶解法制备鸽血红蛋白抗氧化肽的最优工艺条件。结果表明,蛋白酶K为虚拟筛选出的最佳蛋白酶,最佳的制备鸽血红蛋白抗氧化肽的工艺为鸽血红蛋白浓度1 g/100 mL、pH8、酶解时间3 h、酶添加量415.54 U/g、酶解温度65℃。在此条件下,通过验证试验得出响应值DPPH自由基清除率为(33.21±0.68)%,与预测值相差较小,故而此条件为最优工艺。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳试验,发现最优工艺条件下鸽血红蛋白酶解后目的条带消失,进一步验证该方法为酶解鸽血红蛋白源抗氧化肽的最优工艺。
Pigeon hemoglobin was used as raw material to simulate enzymolysis by using proteases in the online database ExPASy Peptide Cutter,and the potential biological activity,solubility,toxicity,and sensitization of peptides were filtered by in silico tools to select the most suitable proteases for preparing pigeon hemoglobin-derived antioxidant peptides. Different enzymolysis time,enzyme addition,and enzymolysis temperature were set as single factors,and the effects of different enzymolysis conditions on the hydrolysis degree and 1,1-diphenyl-2-picrylhydrazyl(DPPH) radical scavenging rate were studied. Based on the single-factor experiment,the optimal process conditions for preparing antioxidant peptides derived from pigeon hemoglobin by enzymolysis were explored by response surface methodology. The results indicated proteinase K was the optimal protease for the preparation of pigeon hemoglobin antioxidant peptides by enzymolysis,and the optimal conditions were pigeon hemoglobin concentration of 1 g/100 mL,pH8,enzymolysis time of 3 h,enzyme dosage of 415.54 U/g,and enzymolysis temperature of 65 ℃. Under these conditions,through the validation test,the response value of the DPPH radical scavenging rate was(33.21±0.68)%,which was consistent with the predicted value. Therefore,the optimal processing conditions for enzymolysis of antioxidant peptides from pigeon hemoglobin were obtained.Moreover,the sodium laurylsulfonate-polyacrylamide gel electrophoresis(SDS-PAGE) test was used to find that the target band disappeared after the enzymolysis of pigeon hemoglobin under the optimal process conditions,which further confirmed this method as the optimal process for enzymolysis of pigeon hemoglobin-derived antioxidant peptides.
作者
力俊琛
高昕悦
江晨怡
杨云顺
赛买提·艾则孜
刘雅娜
巴吐尔·阿不力克木
LI Jun-chen;GAO Xin-yue;JIANG Chen-yi;YANG Yun-shun;Saimaiti Aizeizi;LIU Ya-na;Batuer Abulikemu(College of Food Science and Pharmacy,Xinjiang Agricultural University,Urumqi 830052,Xinjiang,China)
出处
《食品研究与开发》
CAS
北大核心
2023年第2期108-115,共8页
Food Research and Development
基金
新疆维吾尔自治区科技成果转化示范专项(2020C005)。
关键词
鸽血红蛋白
抗氧化肽
模拟酶解
响应面分析法
十二烷基硫酸钠-聚丙烯酰胺凝胶电泳
pigeon hemoglobin
antioxidant peptides
virtual enzymolysis
response surface analysis
sodium laurylsulfonate-polyacrylamide gel electrophoresis(SDS-PAGE)
