牦牛血抗氧化肽发酵制备工艺优化及结构分析研究

Study on the optimization of fermentation preparation process and structural of antioxidant peptides of yak blood

中文关键词:  牦牛血  发酵  抗氧化肽  蛋白质  结构

英文关键词:yak blood  fermentation  antioxidant peptide  protein  structure

中文摘要:

      目的 优化牦牛血抗氧化肽的发酵制备工艺并分析其结构。方法 在单因素实验的基础上, 开展牦牛血抗氧化肽制备的响应面优化实验。采用液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry, LC-MS/MS)对经冷冻干燥处理后的牦牛血抗氧化肽的肽段组成及其序列进行检测。结果 牦牛血抗氧化肽制备的最佳工艺条件为: 底物质量浓度65 g/L、接种量2.0%、发酵时间71 h。在此条件下, 牦牛血抗氧化肽的羟基自由基清除率为88.21%。根据其氨基酸组成、肽段序列、分子量大小筛选出可能具有较好抗氧化活性的肽段20条, 其所匹配的蛋白为A0A6B0QV06、A0A6B0QW84。经蛋白质数据库Uniprot鉴定分析, 此两类蛋白质已形成由两条α链和两条β链组成的异四聚体结构, 可促进动物肺脏器官中的氧气运输到其他脏器组织中, 具有抗缺氧的作用。结论 采用枯草芽孢杆菌液态发酵法制备牦牛血抗氧化肽, 可增大牦牛血的利用率, 减少资源的浪费。

英文摘要:

      Objective To optimize the fermentation preparation process of yak blood antioxidant peptide and analyze its structure. Methods The response surface optimization test was conducted on the basis of single factor test, liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to detect the peptide composition and sequence of antioxidant peptide of yak blood after freeze-drying treatment. Results The optimum technological conditions for the preparation of antioxidant peptides from yak blood were as follows: Substrate mass concentration 65 g/L, inoculation amount 2.0%, fermentation time 71 h. Under these conditions, the scavenging rate of hydroxyl radical was 88.21%. According to its amino acid composition, peptide sequence and molecular weight, 20 peptides with better antioxidant activity were screened out, and the matching protein was A0A6B0QV06 and A0A6B0QW84. According to the identification and analysis of the protein database Uniprot, these 2 kinds of proteins had formed heterotetramer structure which was consisting of two α chain and two β chain. The two types of proteins which could promote the transportation of oxygen from animal lung organs to other organs and tissues had the effect of anti-hypoxia. Conclusion Using Bacillus subtilis liquid fermentation to prepare antioxidant peptides from yak blood can improve the utilization rate of yak blood, and reduce the waste of resources.