大孔吸附树脂纯化苦荞多肽工艺研究

Purification of Tartary buckwheat polypeptide by macroporous adsorption resin

【目的】研究苦荞多肽纯化工艺和抗氧化活性。【方法】以大孔吸附树脂吸附率和解吸率为考察指标,对5种大孔吸附树脂(AB-8、D101、HPD600、ZCX-11和S-8)进行筛选,通过动态吸附-解吸单因素试验和正交试验优化大孔吸附树脂最佳纯化工艺,并测定大孔吸附树脂纯化前后苦荞多肽的超氧化物歧化酶活力、2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐和1,1-二苯基-2-苦基肼清除能力。【结果】从5种大孔吸附树脂中筛选出S-8为最佳大孔吸附树脂,其吸附率和解吸率分别为71.25%、89.00%;在上样质量浓度为5 mg/mL,洗脱流速为1.5 mL/min,乙醇体积分数为55%的条件下,苦荞多肽纯度由粗提液的22.18%提高到70.00%;纯化后苦荞多肽超氧化物歧化酶酶活力显著增加,从原来的26.20 U/mL升高到40.37 U/mL,对2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐和1,1-二苯基-2-苦基肼抑制率显著升高,分别是粗提液的2倍与1.4倍。【结论】优化后的工艺条件可使苦荞多肽的纯度显著提高,且纯化后苦荞多肽抗氧化活性显著增强。

【Objective】The purification process and antioxidant activity of Tartary buckwheat polypeptide were studied to provide basic reference for its production and application.【Methods】Five kinds of macroporous adsorption resins(AB-8,D101,HPD600,ZCX-11,S-8)were screened by taking the adsorption rate and resolution rate of macroporous adsorption resin as the investigation indexes.The optimal purification process of macroporous adsorption resin was optimized by dynamic adsorption-resolution single factor experiment and orthogonal experiment.Superoxide dismutase(SOD)activity,2,2-biazo-bis(3-ethyl-benzothiazol-6-sulfonic acid)ammonium salt(ABTS free radical)and 1,1-diphenyl-2-picrylhydrazine(DPPH free radical)scavenging activity of Tartary buckwheat polypeptides were determined before and after purification by macroporous resin.【Results】S-8 macroporous adsorption resin was selected as the best adsorption resin from the five kinds of resins,and its adsorption rate and resolution rate were 71.25%and 89.00%.Under the conditions of loading concentration of 5 mg/mL,elution flow rate of 1.5 mL/min and ethanol concentration of 55%,the purity of Tartary buckwheat polypeptide was increased from 22.18%to 70.00%.After purification,SOD activity of Tartary buckwheat polypeptide increased significantly from 26.20 U/mL to 40.37 U/mL,and the inhibition rates of DPPH and ABTS radical were increased significantly,which were 2 times and 1.4 times of the crude extract.【Conclusion】The purity of Tartary buckwheat polypeptide was increased under the optimized conditions,and the antioxidant activity of Tartary buckwheat polypeptide was significantly enhanced after purification.