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High-F-ratio leech oligopeptide, and enzymatic hydrolysis preparation method and application thereof

A leech and oligopeptide technology, which is applied in the preparation method of peptides, from leech inhibitors, applications, etc., can solve the problems of single product type and forgetting, and achieve the effect of simple preparation process and easy intestinal absorption.

Active Publication Date: 2014-08-27
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Several centuries ago, humans have successfully used leeches to prevent and treat related diseases. However, with the continuous development of medical technology, drugs that can have the same curative effect, such as aspirin and nitroglycerin, have appeared one after another. forgotten by the world
At present, the leech products on the market are mainly anticoagulant and antithrombotic products, and the product types are relatively single. Therefore, to explore the potential of more abundant applications of leeches, develop a simple method, low cost, and easy absorption of leeches with high F value and low Peptides have a certain market value

Method used

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  • High-F-ratio leech oligopeptide, and enzymatic hydrolysis preparation method and application thereof
  • High-F-ratio leech oligopeptide, and enzymatic hydrolysis preparation method and application thereof
  • High-F-ratio leech oligopeptide, and enzymatic hydrolysis preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0043] Take 500g of dried leeches, clean them, soak them in deionized water and ethanol for 12 hours, put them in an oven at 45°C and dry them until the water content is less than 3%, grind them into fine powder at high speed, add water to dissolve, and the weight ratio of dry powder to water is 5: 100 to prepare the suspension, and homogenize it with a high-speed homogenizer, then adjust the pH to 10 with 1mol / L NaOH, soak for 20min; then adjust the pH to 9, add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the reaction temperature was maintained at 60°C, and the enzymolysis was carried out for 4 hours; boiled for 10 minutes, inactivated, cooled, centrifuged at 8000r / min for 5 minutes, the supernatant was collected, the pH was adjusted to 7.0, and papain was added at 4000U / g pro ( Novozymes Company, Denmark), the reaction temperature was kept at 60°C, and the enzymolysis was carried out for 4 hours; boiled for 10 minutes to inactivate the enzyme, cooled, centrif...

Embodiment 2

[0045] Take 500g of dried leeches, clean them, soak them in deionized water and ethanol for 10 hours, put them in an oven at 45°C and dry them until the water content is less than 3%, grind them into fine powder at high speed, add water to dissolve, and the weight ratio of dry powder to water is 8: 100 to prepare the suspension, and homogenize it with a high-speed homogenizer, then adjust the pH to 11 with 1mol / L NaOH, soak for 50min; then adjust the pH to 10, add Alcalase2.4L alkaline protease (Novozymes company, Denmark), the reaction temperature was maintained at 50°C, and the enzymolysis was carried out for 3 hours; boiled for 20 minutes, inactivated, cooled, centrifuged at 6000r / min for 10 minutes, the supernatant was collected, the pH was adjusted to 8.0, and papain was added at 6000U / g pro ( Novozymes Company, Denmark), the reaction temperature was kept at 60°C, and the enzymolysis was carried out for 3 hours; boiled for 15 minutes to inactivate the enzyme, cooled, centr...

Embodiment 3

[0047] Take 500g of dried leeches, clean them, soak them in deionized water and ethanol for 15 hours, put them in an oven at 45°C and dry them until the water content is less than 3%, grind them into fine powder at high speed, add water to dissolve, and the weight ratio of dry powder to water is 2: Prepare the suspension at 100, and homogenize with a high-speed homogenizer, then adjust the pH to 8 with 1mol / L NaOH, and soak for 60 minutes; then adjust the pH to 8.5, add Alcalase2.4L alkaline protease ( Novozymes company, Denmark), keep the reaction temperature at 65°C, enzymolysis for 3.5h; boil for 10min, inactivate the enzyme, cool, centrifuge at 5000r / min for 20min, collect the supernatant, adjust the pH to 7.0, add papaya at 5000U / g pro Protease (Novozymes, Denmark), the reaction temperature was kept at 55°C, and the enzymolysis was carried out for 2.5 hours; boiled for 10 minutes, inactivated, cooled, centrifuged at 8000r / min for 10 minutes, and the supernatant was collect...

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Abstract

The invention discloses high-F-ratio leech oligopeptide and an enzymatic hydrolysis preparation method and application thereof. The preparation method comprises the following steps: (1) preliminarily cleaning dried leech bodies, removing impurities and carrying out high-speed crushing; (2) pre-soaking raw materials with alkali; (3) carrying out enzymatic hydrolysis by using double enzymes consisting of alkali protease AlcalaseAF 2.4L and papain; (4) statically adsorbing and removing aromatic amino acids by using active carbon powder; and (5) carrying out further separation and purification by using a gel filtration chromatographic column, collecting a separation product and then carrying out freeze drying so as to obtain leech oligopeptide powder with a molecular weight of less than 2000 Da and an F ratio (Fishcer ratio, i.e., a ratio of the mole content of branched-chain amino acids to the mole content of aromatic amino acids) of more than 20. According to the invention, preparation process is simple, two commercial enzymes consisting of alkali protease and papain are selected and used, industrial production can be easily realized, high yield is obtained, and the high-F-ratio leech oligopeptide has a certain anticoagulation effect and increases a novel source for high-F-ratio products.

Description

technical field [0001] The invention relates to a biological peptide and its preparation method and application, in particular to a leech high F value oligopeptide and its enzymolysis preparation method and application. Background technique [0002] High F value oligopeptide is a biologically active oligopeptide mixture with a molar ratio of branched-chain amino acids and aromatic amino acids greater than 20. It can not only relieve the symptoms of hepatic encephalopathy, improve liver function and the degree of hepatic coma, but also have a positive effect on patients with protein malnutrition. In actual production, high F value oligopeptides can be widely used in drugs for the treatment of liver diseases, liver protection food, protein nutrition food for surgical patients, intestinal nutrition for patients with digestive enzyme deficiency and food nutrition fortification for high-intensity laborers agents etc. The high F oligopeptide mixture has more practical utility th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/815C12P21/06A23L1/29C07K1/18A23L33/00
CPCA23L33/18C07K14/815
Inventor 吴晖任尧赖富饶杨祎静李晓凤
Owner SOUTH CHINA UNIV OF TECH
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