Separation and preparation method of rice bran antioxidant peptide

A technology of antioxidant active peptides and rice bran, which is applied in the preparation method of peptides, chemical instruments and methods, peptides, etc., can solve the problems that the research field is still in the blank stage, and achieve the effect of improving the utilization of by-products and saving production costs

Active Publication Date: 2017-05-10
长沙飞创生物技术有限责任公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] After investigation, there is no relevant literature report on Chinese scholars using neutral protease to extract antioxidant active peptides from rice bran gluten.

Method used

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  • Separation and preparation method of rice bran antioxidant peptide
  • Separation and preparation method of rice bran antioxidant peptide
  • Separation and preparation method of rice bran antioxidant peptide

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0028] The method for separating and preparing the rice bran antioxidant active peptide comprises the following steps:

[0029] (1) The technological process of extracting rice bran gluten by alkali extraction and acid precipitation method is: defatted rice bran (through a 60-mesh sieve) → add water and adjust the pH to 9.0 (the ratio of solid to liquid is 1:10, and the unit of the ratio of solid to liquid is mL / g, water bath: 40°C, 4h)→centrifugation (8000r / min, 15min)→take the supernatant→acid precipitation (pH 4.0)→centrifugation (8000r / min, 15min)→take the precipitate→wash three times, adjust the pH7.0 → Freeze-drying (48h) → Alkaline extraction of rice bran gluten (sealed storage at 4°C + silica gel) → Osborne fractional extraction to obtain relatively pure gluten;

[0030] (2) Choose Na 2 CO 3 -NaHCO 3 (pH10.0, 0.1mol / L) was used as a buffer for preliminary separation of rice gluten. With 0.05mol / L Na 2 HPO 4 -NaOH (pH12.0) buffer to dissolve the alkaline-extracte...

Embodiment 2

[0038] Example 2 Detection of Antioxidant Activity of Rice Glutenin Active Peptide in vivo

[0039] Select the cells in the logarithmic growth phase, digest them with EDTA-free trypsin to make a cell suspension, measure the cell concentration in the suspension with a counting plate, inoculate 5000 cells in a 96-well plate, inoculate 100 μL in each well, and inoculate 100 μL in each well. Set up 6 replicate wells, at 37°C 5% CO 2 After being cultured in the incubator for 6 hours, they were divided into normal group, H 2 o 2 Treatment group, H 2 o 2 + Rice glutenin active peptide protection group, suck out the original culture medium, all groups first add 80 μL culture medium, normal group, H 2 o 2 Add 10 μL of serum-free medium to the treatment group, H 2 o 2 + Rice glutenin active peptide protection group was added with corresponding concentration and equal volume of rice glutenin active peptide, so that the final concentration of the drug was 100, 150, 200 μmol / L respe...

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Abstract

The invention provides a bioactive peptide derived from rice bran glutelin, wherein the sequence of the bioactive peptide is shown as KHNRGDEF. The active peptide provided by the invention, which has an antioxidant activity, can be used for preparing foods and health care products capable of enhancing antioxidant efficacy. A polypeptide, which has an antioxidant activity in vitro, is separated from an enzymatic hydrolysate of the rice bran glutelin; an antioxidant active peptide product is prepared by extracting crude glutelin by virtue of a method of alkaline extraction and acid deposition, then conducting sequential extraction, separating and purifying the rice bran glutelin by virtue of ion-exchange chromatography and gel chromatography technologies, hydrolyzing obtained rice bran glutelin by virtue of alkaline protease, and conducting centrifuging, vacuum concentration and freeze-drying, wherein the rice bran glutelin is enzymatically hydrolyzed under the following conditions: an [E] / [S] ratio is 1.5% 1.8%, a pH value is 10.0, a temperature is at 50 DEG C and a duration is 4.2h; and a required antioxidant active peptide pure product is prepared by conducting ion-exchange chromatography and gel chromatography, preparing RP HPLC and analytical RP HPLC, and conducting separating and purifying. A separation and preparation method provided by the invention is low in production cost, simple and convenient in process and is green and environmentally friendly; the prepared rice bran active peptide has the characteristics of being reasonable in amino acid composition, low in allergy, safe and reliable, and the like; and the rice bran active peptide is relatively good in free radical scavenging capacity and excellent in antioxidant efficacy.

Description

technical field [0001] The invention relates to a method for separating and preparing rice bran antioxidant active peptide. Background technique [0002] Rice bran is a by-product produced more in rice production factories. The amino acid composition of its protein is similar to that in rice. The amino acid composition is reasonable, and it is a high-quality protein. A large amount of rice bran is produced every year in my country's industrial production, and most of it has not been effectively utilized. Therefore, it is necessary to further process rice bran protein to increase its added value and realize full utilization of resources. The protein solubility in rice and rice bran is poor, which seriously affects its application. Peptides with smaller molecular weights in rice bran have better solubility, and different peptides have different effects, such as antioxidant, blood pressure lowering, immune activity, etc. Rice antioxidant active peptide has good free radical s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K14/415C07K1/18C07K1/16C07K1/20A61P39/06
CPCC07K14/415C12P21/06
Inventor 梁盈林亲录王玉倩李佳佳黄萍张琳
Owner 长沙飞创生物技术有限责任公司
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