带鱼黄嘌呤氧化酶抑制肽制备及工艺优化

Preparation and process optimization of xanthine oxidase inhibitory peptide from Trichiurus lepturus

中文关键词:  带鱼  酶解  肌肽  鹅肌肽  黄嘌呤氧化酶

英文关键词:Trichiurus lepturus  enzymolysis  carnosine  anserine  xanthine oxidase

中文摘要:

      目的 制备带鱼黄嘌呤氧化酶抑制肽, 研究含肌肽类活性肽的带鱼提取物对黄嘌呤氧化酶的抑制作用。方法 以带鱼为原料, 通过超声辅助酶法制备带鱼黄嘌呤氧化酶抑制肽, 高效液相色谱法测定肌肽类活性肽的含量。结果 以肽粉得率和黄嘌呤氧化酶抑制率为指标, 筛选得到碱性蛋白酶为最佳反应用酶。以单因素实验为基础, 通过Box-Behnken响应面分析法, 以水解度、黄嘌呤氧化酶抑制活性、肌肽及鹅肌肽含量为响应值, 优化得到酶解最优条件: 酶解时间5 h, 超声时间21 min, 酶解温度49℃。在此条件下, 得出水解度21.90%、黄嘌呤氧化酶抑制率52.03%、鹅肌肽含量0.66%和肌肽含量为0.15%, 与回归理论模型基本符合。在最优酶解方案的基础上, 将酶解液进一步分成不同组分, 可得到肌肽类活性肽含量升高, 抑制活性变大。结论 肌肽类活性肽对黄嘌呤氧化酶活性的抑制有着积极的作用, 这为工业化利用带鱼制备黄嘌呤氧化酶抑制肽提供理论及指导。

英文摘要:

      Objective To prepare xanthine oxidase inhibitory peptides from Trichiurus lepturus, and study the inhibitory effect of Trichiurus lepturus extract containing carnosineactive peptides on xanthine oxidase. Methods The inhibition peptide of xanthine oxidase of Trichiurus lepturus was prepared by ultrasound-assisted enzymatic method, and the content of carnosine-like active peptide was determined by high performance liquid chromatography. Results The peptide powder yield and xanthine oxidase inhibition rates were used as indicators to screen the alkaline protease as the best enzyme for the reaction. Based on the single-factor experiment, the optimal conditions for enzymatic digestion were optimized by Box-Behnken response surface analysis with the degree of hydrolysis, xanthine oxidase inhibition activity, carnosine and anserine content as the response values: The enzymolysis time was 5 h, the sonication time was 21 min, and the enzymatic digestion temperature was 49℃. Under these conditions, the hydrolysis degree of 21.90%, xanthine oxidase inhibition rate of 52.03%, anserine content of 0.66% and carnosine content of 0.15% were obtained, which were basically consistent with the regression theoretical model. On the basis of the optimal enzymatic solution, the enzymatic solution was further divided into different fractions, and the carnosine active peptide could be obtained to have a positive effect on the inhibition of xanthine oxidase activity. Conclusion Carnosine active peptides have a positive effect on the inhibition of xanthine oxidase activity, which provides theory and guidance for the industrial preparation of xanthine oxidase inhibitory peptides by Trichiurus lepturus.