High throughput screening system, screening method and preparation method for chlorella functional component CPE

A functional ingredient, chlorella technology, applied in biochemical equipment and methods, material excitation analysis, microbial determination/inspection, etc., can solve problems such as wall breaking, and achieve the effect of high functional activity and wide range of activity spectrum.

Inactive Publication Date: 2017-03-15
EAST CHINA UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

For example, due to the existence of tough cell walls, there is a problem of whether the active ingredients extracted from Chlorella are broken

Method used

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  • High throughput screening system, screening method and preparation method for chlorella functional component CPE
  • High throughput screening system, screening method and preparation method for chlorella functional component CPE
  • High throughput screening system, screening method and preparation method for chlorella functional component CPE

Examples

Experimental program
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Effect test

Embodiment 1

[0081] A high-throughput activity screening system for CPE, a functional component of Chlorella algae

[0082] 1) Antioxidant test: Griess reagent is used for testing, and the total antioxidant capacity and the scavenging effect on free radicals of the sample are detected by spectrophotometry;

[0083] 2) Cellular immune regulation test:

[0084] a. The effect of tetrathiazolium blue MTT assay components on the survival rate of mouse macrophage Ana-1;

[0085] b. Neutral red NR staining method to detect the effect of components on Ana-1 phagocytosis;

[0086] c. Fluorescence probe method was used to detect the change of NO secretion in Ana-1 cells by samples;

[0087] 3) Redox metal ion chelation test:

[0088] a. Using ferrozine as Fe 2+ Chelating reagent, detection of Fe in samples by spectrophotometry 2+ Chelation ability;

[0089] b. Using catechol violet as Cu 2+ Chelating reagent, Cu detection of samples by spectrophotometry 2+ Chelation ability;

[0090] 4) Inh...

Embodiment 2

[0094] In this example, CPE1 and CPE2 were selected to be prepared by ultrasonic enzymolysis, and the specific method was as follows: 10 g of Chlorella pyrenoidosa powder was evenly suspended in 100 mL of distilled water. After ultrasonically disrupting the wall for 30 min, extract in a water bath at 95°C for 30 min. After the extract is cooled, cellulase (80 mg / g algae powder) and pectinase (10 mg / g algae powder) are added. Degrade the polysaccharide in a water bath at 50°C for 2 hours. Boiling water bath for 30min to inactivate the enzyme, centrifuge at 10000rpm for 30min. The supernatant was collected, concentrated by 4 times volume, and freeze-dried to obtain CPE sonication hydrolysis products, wherein CPE1 was hydrolyzed for 4 hours, and CPE2 was hydrolyzed for 16 hours.

[0095] A high-throughput activity screening method for CPE, a functional component of Chlorella algae

[0096] 1) Antioxidant ability test, that is, the ability to scavenge hydroxyl radicals (˙OH)

...

Embodiment 3

[0126] Method for preparing chlorella functional component CPE based on high pressure wall breaking method

[0127] Take 10g of Chlorella pyrenoidosa algae powder and suspend evenly in 100mL of distilled water. Place in a pressure cooker, heat at 0.15MPa, 120°C for 40min. The samples were taken out and cooled at room temperature. Centrifuge at 10,000 rpm for 30 min, and the supernatant is filtered under reduced pressure and stored at 4°C for later use. Add 100mL of distilled water to resuspend the centrifuged algae mud, place in a pressure cooker, heat at 0.15MPa, 120°C for 40min, and extract again. The samples were taken out and cooled at room temperature. Centrifuge at 10,000 rpm for 30 min, take the supernatant, and filter under reduced pressure. The two filtrates were combined and freeze-dried to obtain a CPE extract powder, labeled as CPE-a.

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Abstract

The invention relates to a high throughput screening system, screening method and preparation method for a chlorella functional component CPE. The screening method comprises 1) an antioxidant test, 2) a cell immunoregulation test, 3) a redox metal ion chelation test, and 4) an advanced glycation end product AGEs inhibition test. The preparation method comprises adding chlorella pyrenoidosa powder into distilled water, heating the mixture at 110-130 DEG C for 30-60min under high pressure, carrying out extraction, taking out a sample, cooling the sample at the room temperature, carrying out centrifugation at a rotation rate of 10000r for 30min, carrying out vacuum suction filtration on the supernatant, storing the filtrate at 4 DEG C for nest use, adding the same amount of distilled water into the chlorella mud obtained by centrifugation, carrying out resuspension, repeating the above processes, merging the filtrates obtained by the two processes, and carrying out freeze drying to obtain chlorella CPE extract powder. The preparation method has a high yield of the chlorella functional component CPE, and the extract is rich in nucleic acid and proteins. The preparation method is suitable for industrial production, reduces a development cost of microalgae energy and improves the comprehensive utilization degree of microalgae energy.

Description

technical field [0001] The invention belongs to the technical field of screening, preparation and extraction of CPE, a functional component of chlorella, and particularly relates to a high-throughput screening system and screening method for CPE, a functional component of chlorella, and a method for preparing CPE, a functional component of chlorella, based on a high-pressure wall-breaking method. method. Background technique [0002] Due to the advantages of short growth cycle, high biomass yield, strong carbon dioxide fixation capacity, high production efficiency per unit of land and no competition with food, microalgae are gradually surpassing traditional energy crops and becoming the third generation of new generation bioenergy. Although the research on the industrialization of microalgae energy has entered the pilot stage, there are still many difficulties, the most important of which is the high cost. [0003] Chlorella (Chlorella), commonly known as green algae, is on...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N21/64C12Q1/02
Inventor 陶黎明张道敬李元广庄秀园秦文贾敬
Owner EAST CHINA UNIV OF SCI & TECH
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