双酶法酶解豌豆蛋白制备高抗氧化多肽的研究

Study on Preparation of Antioxidant Peptides from Pisum sativum L. by Dual-enzymatic Method

以水解度及DPPH自由基清除能力为指标优选出1398中性蛋白酶和胰蛋白酶分步酶解豌豆分离蛋白,所得豌豆蛋白水解产物(pea protein hydrolysate,PPH)中相对分子量小于2 KDa的多肽占77.07%,对DPPH的半清除浓度(IC50)为3.01 mg/mL;PPH经Sephadex G-25凝胶层析和两次RESOURCETM3RPC反相层析纯化获得高抗氧化活性峰,经反相层析鉴定为层析纯,当浓度在0.2 mg/mL时,对DPPH的清除率达到62.03%,与同浓度下的谷胱甘肽对DPPH的清除率相近(66.82%)。

In this paper, 1398 neutral protease and trypsin were screened to prepare pea protein hydrelysate （PPH） by using the degree of hydrolysis and DPPH radicals scavenging activity as indexes. The less than 2 KDa fraction of PPH accounted for 77.07% and the concentration of PPH required to scavenge 50% of DPPH radicals （ ICs0 ） was 3.01 mg/ mL. Antioxidant peptides were isolated by gel filtration chromatography on Sephadex G-25 and reversed phase chroma- tography on RESOURCETM 3RPC. The purity of antioxidant peptides was analyzed by reversed phase chromatography. The results showed that DPPH radicals scavenging activity of antioxidant peptides could reach 62.03% at the concentrations of 0.2 me-/mL,close to lutathione （66.82%）.