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Preparation method for chlorella anti-oxidative peptide

A technology of antioxidant peptides and chlorella, applied in biochemical equipment and methods, methods based on microorganisms, microorganisms, etc., to achieve high efficiency, strong antioxidant activity, and high purity

Active Publication Date: 2014-10-29
安徽汉芳生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Research on the extraction of natural antioxidant drugs from marine algae to be carried out

Method used

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  • Preparation method for chlorella anti-oxidative peptide
  • Preparation method for chlorella anti-oxidative peptide
  • Preparation method for chlorella anti-oxidative peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] 1 Extraction process of chlorella protein

[0041] 1.1 Materials and reagents

[0042] Chlorella dry powder, from Hangzhou Huadan Agricultural Products Co., Ltd., Zhejiang Province; Bio-Rad Protein Assay kit, purchased from Bio-Rad Laboratories Inc; copper sulfate, potassium sulfate, sulfuric acid, boric acid, sodium hydroxide, 95% ethanol, mixed Indicator solution (2 parts of methyl red ethanol solution and 1 part of methylene blue ethanol solution are mixed immediately before use).

[0043] 1.2 Experimental Instruments

[0044] Enzyme-linked immunoassay instrument (Model-550), American Bio-Rad company; High-speed refrigerated centrifuge (5415R), German Eppendorf company; Electronic balance (BS224S); Kjeldahl nitrogen analyzer (KDY-9820) AL04 electronic balance, HH-2 water bath, DHG-9240A blast drying oven, DL-5M centrifuge (with replacement rotor), SANYO ultra-low temperature refrigerator.

[0045] 1.3 Experimental method

[0046] 1.3.1 Determination of protein co...

Embodiment 2

[0114] Take 20g of chlorella powder, add 400ml of distilled water at a material-to-liquid ratio of 1:20 (m / v), and stir for 1 hour at 4°C; slowly put the chlorella suspension into an ultra-low temperature refrigerator, and freeze at -70°C 12h; then thaw at 30°C to 4°C, slowly add NaOH to the thawed chlorella mixture to a final concentration of 1%, and keep stirring to obtain an alkaline mixture; at 4°C, the alkali The mixed solution was ultrasonically broken for 60 minutes, and the ultrasonic condition was 30KHz, and then the pH was adjusted to 7.0 with 0.5M hydrochloric acid; centrifuged at 4000rpm for 20 minutes, and the supernatant was taken; 95% ethanol solution with a volume fraction of 4 times the volume of the supernatant was added , continuously stirred for 1 min, and stood still for 5 h; centrifuged at 2000 rpm for 10 min, discarded the supernatant, dissolved the precipitate in 50 mL of distilled water, concentrated it to about 30 mL, and obtained 2.26 g of chlorella p...

Embodiment 3

[0121] Take 20g of chlorella powder, add 400ml of distilled water at a material-to-liquid ratio of 1:20 (m / v), and stir for 1 hour at 4°C; slowly pour the chlorella suspension into a high-pressure homogenizer, and adjust the pressure to 100MPa , repeated 20 times, and then in 4°C environment, slowly add NaOH to the chlorella mixed solution after high-pressure homogenization treatment to a final concentration of 1%, and keep stirring to obtain an alkaline mixed solution; Under normal circumstances, the alkaline mixed solution was treated with ultrasonic wall breaking for 60min, and the ultrasonic condition was 30KHz, and then the pH was adjusted to 7.0 with 0.5M hydrochloric acid; centrifuged at 4000rpm for 20min, and the supernatant was taken; a volume fraction of 4 times the volume of the supernatant was added to 95 % ethanol solution, continuously stirred for 1 min, and stood still for 5 h; centrifuged at 2000 rpm for 10 min, discarded the supernatant, dissolved the precipita...

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Abstract

The invention discloses a preparation method for a chlorella anti-oxidative peptide. The preparation method is as follows: by taking chlorella as material, carrying out wall-breaking treatment on the chlorella to obtain soluble protein by utilizing a dilute alkali swelling-combined ultrasonic-wave process; and hydrolyzing the chlorella protein by a biological enzyme method to obtain the chlorella anti-oxidative peptide with remarkable anti-oxidative effect. The invention further discloses a treatment process combining ultralow temperature treatment, high pressure treatment with dilute alkali swelling and ultrasonic waves, which can improve yield of the water-soluble protein and can further improve anti-oxidative ability of hydrolysate. A product obtained by virtue of the preparation method disclosed by the invention is high in purity and strong in anti-oxidative activity.

Description

(1) Technical field [0001] The invention relates to a preparation method of chlorella antioxidant peptide. (2) Background technology [0002] Free radicals are intermediate metabolites of various biochemical reactions in human life activities. Due to their single-electron structure, they show extremely active instability. In organisms, they mainly include superoxide anion radicals (O 2 - ), hydrogen peroxide (H 2 o 2 ), hydroxyl radical ( OH), oxygen organic radical (RO ), organic peroxy radical (ROO ), reactive oxygen species (ROS), lipid radical (L ) and lipoperoxy radical (LOO ) Wait. Active free radicals can cause varying degrees of damage to general tissues and biofilms. Therefore, the mechanism of free radicals in physiology and pathology has received extensive attention, and more and more scholars have devoted themselves to related research. [0003] There are two main sources of oxygen free radicals in the human body: exogenous and endogenous. "Exogenous free r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C12R1/89
Inventor 张拥军王爽张高帆苏东洋朱丽云
Owner 安徽汉芳生物科技有限公司
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