Study On Process Of Enzymatic Hydrolysis Chinese Pine Pollen Protein For Preparing Small Peptide

In the paper ethanol,n-hexane and water as extraction solvent to study the different conditions for extraction of flavonoids,unsaturated fatty acids and total output,drawn in ethanol:n-hexane = 7:3 and 1:1 extract flavonoids and the total output being better,and the conditions of the composite extract for the single factor was in research,the optimum conditions were:(1)m(ethanol):m(n-hexane)=7:3,the temperature was 50℃,the reaction time of 3h,solid-liquid ratio of 1:10;(2)m(ethanol):m(n-hexane)=1:1,the temperature was 50℃,the reaction time of 3h,solid-liquid ratio of 1:20.The unsaturated fatty acids of the composite extraction determined qualitatively by Liquid Chromatography.The design also selected temperature,the ethanol concentration,the ratio solid-liquid in optimization the flavonoids extraction conditions.The optimum conditions is:solvent is 70%ethanol,temperature is70℃,the ratio is 1:20.In these conditions,the content of flavonoids was 1.41%by NaNO₂-Al(NO₃)₃-NaOH colorimetric method.Response surface design(RSD) was applied to find the optimum extracting conditions for total flavonoids from Chinese pine pollen.The optimum conditions of extraction were as follows: temperature is 80℃,the addition of solvent(56%ethanol) was 17.9 times the addition of solute,then reflux in heated water-bath 3.2h.In these conditions,the content of flavonoids was 1.49%.Determination of total phosphorus in Chinese pine pollen by phosphorus measurement,the content was 0.66%.The paper used alkaline protease,papain and double emzymatic hydrolysis prepared small peptide of pine pollen.Hydrolyzed by alkaline protease:temperature is 50℃, substrate concentration is 4.5%,pH is 10,hydrolyzing time is 2 h,and enzyme concentration is 4000 u/g.In these conditions,nitrogen solubility index(NSI) of hydrolyzate is respectively 80.30%.Hydrolyzed by papain:temperature is 45℃,substrate concentration is 4.5%,pH is 6.5,hydrolyzing time is 2 h,and enzyme concentration is 3000 u/g.In these conditions,nitrogen solubility index(NSI) of hydrolyzate is respectively 41.63%.Hydrolyzed by double emzyme:temperature is 45℃,substrate concentration is 4.5%,pH is 9,hydrolyzing time is 2 h,and the proportion of alkaline protease and papain is 1:1.In these conditions,nitrogen solubility index(NSI) of hydrolyzate is respectively 69.75%.Determination of cleaning hydroxygen free radical capacity of hydrolyzate.The inhibition rate of alkaline protease hydrolyzate was 40.57%.The inhibition rate of papain hydrolyzate was 29.25%.The inhibition rate of double emzyme hydrolyzate was 74.84%. Determination of antioxidant activity of hydrolyzate.The antioxidant activity of alkaline protease hydrolyzate was 0.068mg/ml.The antioxidant activity of papain hydrolyzate was 0.034mg/ml.The antioxidant activity of double emzyme hydrolyzate was 0.119mg/ml.