Preparation Of Oviductus Ranae Antioxidant Peptides And Their Activities Component Screening

Oviductus Ranae was female Rana chensinensis’s fallopian tubes dried products that underlow temperature conditions. Because of its’ high nutritional value, it had a long history oftaking.Order to determine the main antioxidant activities component of Oviductus Ranae,Oviductus Ranae were separated.Two sample include water-miscible proteins powder,non-water-miscible ingredients powder were prepared, and their hydroxyl free radicals,superoxide anion free radicals and DPPH· free radicals were determined. Results indicated thatOviductus Ranae’s water-miscible proteins powder has a good antioxidant activity, under thesame conditions, its antioxidant activity was the highest,in10mg/mL concentration,its hydroxylfree radicals scavenging, superoxide anion free radicals scavenging and DPPH· free radicalsscavenging was45.62%,37.8%and71.68%respectively.Water-miscible proteins wasdetermined to be the main antioxidant activities component of Oviductus Ranae.Oviductus Ranae was hydrolyed by papain, neutral protease, and alkaline protease, undertheir optimum hydrolysis conditions.Determination of the antioxidant activities of the threepeptides,in the10mg/mL concentration,the peptides’ prepared of neutral protease hydroxyl freeradicals scavenging, superoxide anion free radicals scavenging and DPPH· free radicalsscavenging was39.31%,32.59%and47%, was the best,so selected the neutral protease to do therest of the experiment. Neutral protease digestion experiments carried out single-factorexperiment and orthogonal experiment, hydroxyl free radicals scavenging as an indicator wasmeasured.The results showed that the best conditions was enzymatic hydrolysis conditionsorthogonal temperature50℃, pH value of7.5, enzyme dosage of4%,and hydrolysis time10h.When the optimum conditions of neutral protease antioxidant peptide prepared at aconcentration of10mg/mL, hydroxyl radical scavenging was39.33%, superoxide anionscavenging was32.61%, DPPH· radical scavenging was47%.Water-miscible proteins, non-water-miscible ingredients was hydrolyed by papain, neutralprotease, and alkaline protease, under their optimum hydrolysis conditions.Determination of thewater-miscible proteins was neutral protease hydrolye was the best,so selected the neutralprotease to do the rest of the experiment.When the optimum conditions of neutral proteaseantioxidant peptide prepared at a concentration of10mg/mL, hydroxyl radical scavenging,superoxide anion scavenging, DPPH· radical scavenging was60.03%,45%and80.17%. Non-water-miscible ingredients hydrolyzed by alkaline protease to improve the itsantioxidant ingredients, but its antioxidant activities after hydrolyzed still be low.Measured by flow cytometry to analysis of the protective effect of water-miscible proteinson hela cells damaged by H2O2. After processing with5×10-4mol/L H2O2,Hela cells rapidlyapoptosis, early apoptosis and late apoptosis rate rose to35.5%and1.4%, the number of livingcells was62.8%.It was found that, H2O2induced apoptosis of hela cells., the number of livingcells was62.8%.Antioxidant peptides of water-miscible improved phenomenon of H2O2inducedapoptosis. When the concentration of antioxidant peptides of water-miscible was2mg/mL,theliving cells’ percentage was63.6%, when the concentration of antioxidant peptides ofwater-miscible increased to the4mg/mL,the living cells’ percentage was71.6%, when theconcentration of antioxidant peptides of water-miscible was6mg/mL,the living cells’ percentagewas78%, when the concentration of antioxidant peptides of water-miscible was8mg/mL and10mg/mL,the living cells’ percentage was84.5%and87.1%. Result indicate that antioxidantpeptides of Oviductus Ranae water-miscible had a protective effect for the H2O2inducedapoptosis.