Method for extracting ACE (Angiotensin Converting Enzyme) inhibitory peptide from urechis unicinctus and application

A single-ring thorn and inhibitory peptide technology, which is applied in the field of comprehensive utilization of biological resources, can solve problems such as separation and purification of active peptides, and achieves the effects of easy mass production, good activity and low production cost.

Inactive Publication Date: 2022-06-24
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the separation and purification of ACE inhibitory active peptides derived from the monocyclic spinyweed

Method used

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  • Method for extracting ACE (Angiotensin Converting Enzyme) inhibitory peptide from urechis unicinctus and application
  • Method for extracting ACE (Angiotensin Converting Enzyme) inhibitory peptide from urechis unicinctus and application
  • Method for extracting ACE (Angiotensin Converting Enzyme) inhibitory peptide from urechis unicinctus and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Preparation of Monocyclic Cysts autolysase

[0027] 1.1 Preparation of the crude enzyme solution of monocyclic thorn autolyase:

[0028] (1) Take the discarded viscera of the single-ringed thorn and thaw at room temperature, which is stored in a freezer at -20--10 °C, and then wash it with sterile water to remove impurities after it is completely thawed. Add PBS (0.1 mol / l, pH 8.0) for homogenization, place at 4 °C overnight, centrifuge at 5000 r / min for 8 min, and collect the supernatant;

[0029] (2) Add ammonium sulfate to the above collected supernatant to 20% saturation, place at 4°C for 4 hours; centrifuge at 8000 r / min for 10 minutes, collect the supernatant, and add ammonium sulfate to the supernatant to 70% Saturation, placed at 4 °C for 4 h; centrifuged at 8000 r / min for 10 min, took the precipitate, added 10 times its volume of PBS buffer to the precipitate, and dialyzed with a dialysis bag with a molecular weight cut-off of 8KD-14KD to obtain aut...

Embodiment 2

[0035] Example 2: Screening of ACE-inhibited peptidolysis target protease

[0036] 2.1 Pretreatment of raw materials: Weigh 1kg of the frozen product of the single-ringed thorn beetle, thaw at 20°C for 3 hours, and then add 5 times the volume of the treatment solution to soak for 30min. The treatment solution consists of 4.5g of sodium chloride, chloramphenicol 0.25g, penicillin 0.25g, EDTA·2Na 0.25g, sterile water 4.09475kg. Homogenize.

[0037] 2.2 Centrifugation of raw materials: add 2% of its mass of enzyme to the homogenate, enzymolysis under optimum conditions, inactivate the enzyme at 100 °C for 15 min; after enzymatic hydrolysis, cool down to 20 °C, centrifuge at 8000 r / min for 20 min, and collect the supernatant by filtration .

[0038] 2.3 Screening of protease: 5 kinds of proteases were added in the slurries respectively, and the ratio of material to liquid was 1:2, wherein, 2% autolysate of monocyclic thorn, pH 7.5, 2% Bacillus licheniformis protease, pH 7.0; 2% ...

Embodiment 3

[0050] Example 3: Optimization of ACE-inhibiting peptide enzymatic hydrolysis process.

[0051] Single factor and response surface process optimization was carried out according to the single-ringed thorn autolysase selected in Example 2.

[0052] 3.1 Pretreatment of raw materials: Weigh 1kg of the frozen product of the single-ringed thorn beetle, thaw at 20°C for 3 hours, and then add 5 times the volume of the treatment solution to soak for 30 minutes. The treatment solution consists of 4.5g of sodium chloride, chloramphenicol 0.25g, penicillin 0.25g, EDTA·2Na 0.25g, sterile water 4.09475kg. Homogenize.

[0053] 3.2 Centrifugation of raw materials: add 2% of its mass enzyme to the homogenate, enzymolysis under optimal conditions, inactivate the enzyme at 100 °C for 15 min; after enzymatic hydrolysis, cool to 20 °C, centrifuge at 8000 r / min for 20 min, and filter to collect the supernatant .

[0054] 3.3 Single factor experiment: determine the ratio of solid to liquid (1:1,...

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Abstract

The invention belongs to the field of comprehensive utilization of biological resources, and particularly relates to a method for extracting ACE (Angiotensin Converting Enzyme) inhibitory peptide from urechis unicinctus and application. The method comprises the following steps: pretreating urechis unicinctus, adding optimized compound protease for enzymolysis, and filtering and collecting supernatant after enzyme deactivation; performing gradient dialysis to obtain a crude solution, performing fine purification by utilizing column chromatography, and performing low-temperature drying to obtain ACE inhibitory peptide dry powder. The urechis unicinctus ACE inhibitory peptide prepared by the method is good in activity, low in cost and easy for mass production, and can be used for formula production of food. Meanwhile, comprehensive high-value utilization of the urechis unicinctus is achieved, and good economic and social benefits are achieved.

Description

technical field [0001] The invention belongs to the field of comprehensive utilization of biological resources, and in particular relates to a method and application of extracting ACE inhibitory peptides from a monocyclic stinging beetle. Background technique [0002] The single-ringed beetle, also known as sea sausage or sea sausage, is a tubular beetle that lives in the muddy bottom of the intertidal zone or in the coral reef area. Naked sea cucumber". The single-ringed stinging beetle has a protein content of 22.84%, is rich in a variety of essential human amino acids, and has a crude fat content as low as 4.24%. It contains polysaccharides, polypeptides, functional proteins and other active components, and has good development prospects in anticoagulation, antitumor, blood pressure, blood lipid, and blood sugar reduction. Therefore, it is of good practical significance to explore the deep processing technology of the single-ringed thorn and extract and utilize the nutr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/36C07K1/34C07K1/16A23L33/18
CPCC12P21/06C07K1/36C07K1/34C07K1/16A23L33/18A23V2002/00A23V2200/326A23V2250/55
Inventor 焦绪栋杨林彤黄栋秦松
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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