青占鱼黄嘌呤氧化酶抑制肽制备工艺优化

Optimization of the preparation process of xanthine oxidase inhibitory peptides from Pneumatophorus japonicus

中文关键词:  青占鱼  酶解  黄嘌呤氧化酶抑制肽  工艺优化

英文关键词:Pneumatophorus japonicus  enzymolysis  xanthine oxidase inhibitory peptide  process optimization

中文摘要:

      目的 优化青占鱼黄嘌呤氧化酶(xanthine oxidase, XOD)抑制肽的制备工艺。方法 以青占鱼为原料, 选用5种蛋白酶对其水解制备XOD抑制肽, 测定XOD抑制率和水解度并用以评价效果, 通过单因素和响应面分析相结合的实验方法优化酶解条件, 最后对其分子量分布情况进行了检测分析。结果 复配酶制剂为最适酶, 最佳酶解工艺为: 加酶量4000 U/g、酶解时间4 h、酶解温度55℃、pH 7.0、料液比1:2 (g/mL); 同时测得最优工艺下XOD抑制率和水解度的实际值分别为68.22%和19.15%, 与理论值基本无差别。采用该法制备的青占鱼XOD抑制肽分子量分布情况为: 5500~3000 Da的肽段占比56.44%, 3000~1500 Da的肽段占比31.07%, <1500 Da的肽段占比12.49%, 均在5500 Da以下, 证明表达生物活性的是小分子肽。结论 本研究可为青占鱼制备XOD抑制肽的生产工艺与机制研究提供一定的技术参考与理论支持。

英文摘要:

      Objective To optimize the preparation process of xanthine oxidase (XOD) inhibitory peptides from Pneumatophorus japonicus. Methods Using Pneumatophorus japonicus as raw materials, the XOD inhibitory peptide was prepared by the hydrolysis of the Pneumatophorus japonicus with 5 kinds of proteases, and the XOD inhibition rates and hydrolysis degrees were measured and used to evaluate the effect. Enzymatic hydrolysis conditions were optimized by combining single factor and response surface analysis. Finally, the distribution of molecular weight was analyzed. Results The mixed enzyme compound was the most suitable enzyme, and the best enzymolysis process: Enzyme dosage was 4000 U/g, enzymolysis time was 4 h, enzymolysis temperature was 55℃, pH was 7.0, material liquid ratio was 1:2 (g/mL). The actual values of XOD inhibition rate and hydrolysis degree were 68.22% and 19.15%, respectively, which were not different from the theoretical values. The molecular weight of XOD inhibitory peptides prepared by this method was as follows: The proportion of 5500?3000 Da peptides was 56.44%, the proportion of 3000?1500 Da peptides was 31.07%, the proportion of peptides <1500 Da was 12.49%. They were all below 5500 Da, which proved that the peptide expressing biological activity was small molecule peptide. Conclusion This study can provide some technical reference and theoretical support for the study of the production process and mechanism of XOD inhibitory peptide preparation in Pneumatophorus japonicus.