Crocodile bone gelatin polypeptide, and preparation method and applications of crocodile bone gelatin polypeptide

A crocodile bone and protease technology, applied in the field of bioengineering, can solve problems such as disorder of taste, excessive reduction of blood pressure, and no research report on crocodile bone glue antihypertensive peptide, etc., and achieves the effect of high degree of hydrolysis, improved activity, and beneficial to human absorption.

CN111777679AInactive Publication Date: 2020-10-16福建鼍龙实业有限责任公司
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2020-10-16
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses crocodile bone gelatin polypeptide. The polypeptide includes, by mass fraction, 80-99% of amino acid, wherein the amino acid includes 5-25% of proline, 0.5-5% of tyrosine and 0.5-8% of phenprobamate. A preparation method of the crocodile bone gelatin polypeptide is also disclosed. The preparation method includes the following steps: cooking crocodile bones of which other tissue such as surplus meat and fat are removed in distilled water, and obtaining crocodile bone gelatin through concentration; performing hydrolysis on the crocodile bone gelatin with protease to obtain enzymatic hydrolysate; centrifuging the enzymatic hydrolysate, taking supernatant to perform freeze drying so as to obtain crocodile polypeptide coarse powder; and separating the crocodile polypeptide coarse powder to obtain the crocodile bone gelatin polypeptide. The prepared crocodile bone gelatin polypeptide is high in degree of hydrolysis and small in molecular weight of peptide fragments; and the IC50 of the crocodile bone gelatin polypeptide on ACE enzyme is within the range from 1.000+ / -0.001 mg / ml to 2.000+ / -0.001 mg / ml.
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Description

technical field

[0001] The application belongs to the technical field of bioengineering, and in particular relates to a crocodile bone glue angiotensin-converting enzyme inhibitory polypeptide, a preparation method and an application. Background technique

[0002] Crocodiles are the oldest reptiles and are close relatives of extinct dinosaurs, so they are known as "living fossils". Crocodiles are full of treasures, such as crocodile meat, which is rich in high-quality protein and essential amino acids, unsaturated fatty acids, vitamins and various trace elements. Crocodile liver has the functions of nourishing the brain, generating new blood, and removing dampness. Eating crocodile liver can detoxify and improve eyesight, nourish blood and kidney, and has obvious therapeutic effects on those with poor liver function and poor eyesight, and can also inhibit the formation of stones. Crocodile bone can prevent and treat rheumatic bone pain, and bone meal contains a large amoun...

Examples

Embodiment 1

[0044] The preparation of embodiment 1 crocodile bone glue ACE inhibitory peptide

[0045] (1) Effects of different proteases on the activity of crocodile bone glue polypeptides

[0046] Remove excess meat and fat from the crocodile bones, wash and chop. Add appropriate amount of distilled water to the processed crocodile bones, the ratio of material to liquid is 1:3, and cook in a pressure cooker for 30-60 minutes. After depressurization, let it stand for cooling to separate the oil and water, remove the upper layer of grease, filter with double-layer gauze, concentrate and dry to obtain the crocodile bone glue. Get 5 parts of equal amounts of crocodile bone glue, add in 5 beakers respectively, add trypsin, papain, neutral protease, alkaline protease, compound protease respectively based on the weight of bone glue 1% in each beaker, respectively in each enzyme At the optimum temperature (the optimum hydrolysis temperature of trypsin is 45°C, the optimum hydrolysis temperatu...

Embodiment 2

[0054] Example 2: Amino acid composition of an angiotensin-converting enzyme inhibitory peptide

[0055] The angiotensin-converting enzyme inhibitory peptide prepared in Example 1 was placed in 6 mol / L HCl, hydrolyzed at 110° C. for 24 hours, and the amino acid composition was determined with an automatic amino acid analyzer. Its amino acid composition is as Image 6 shown. from Image 6 It can be seen that the polypeptide has a complete range of amino acids, and the content accounts for 90.13% of the substance. Among them, the content of proline, tyrosine and phenylalanine is relatively high.

[0056]

Embodiment 3

[0057] Example 3: Study on the inhibition kinetics of ACE by the crocodile bone glue angiotensin-converting enzyme inhibitory polypeptide of the present application

[0058] Enzyme inhibition kinetics is to establish the enzyme kinetics model by detecting the relationship between the substrate concentration change and the reaction rate under different inhibitor concentrations, so as to judge the inhibition mode of the inhibitor. With 0mg / mL, 1mg / mL, and 3mg / mL collagen peptide M3 as inhibitors, the ACE catalytic concentrations of 2mg / mL, 4mg / mL, and 6mg / mL substrates, acryloylphenylalanylglycine The activity of aminoacylglycine. ACE Inhibition Kinetics Study Results See Figure 7 , to obtain a set of parallel lines. The abscissa / S is the reciprocal of the HHL concentration, and the V in the ordinate 1 / V represents the reaction speed, which is measured by the hippuric acid formed after the combination of ACE and HHL. It can be seen that with the increase of the concentration...