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Method for preparing protein peptide with immobilization proteolytic enzyme protolysate

A technology of immobilized protease and immobilized enzyme, applied in the field of producing protein peptides with immobilized protease, can solve the problems of application limitation and high production cost

Inactive Publication Date: 2008-10-29
FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, if free protease is directly added to the protein solution to hydrolyze the protein, the enzyme in the reaction system cannot be recovered, resulting in high production costs and other defects, so its industrial application is limited.

Method used

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  • Method for preparing protein peptide with immobilization proteolytic enzyme protolysate
  • Method for preparing protein peptide with immobilization proteolytic enzyme protolysate

Examples

Experimental program
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Effect test

Embodiment 1

[0020] Using Pluronic P123 as template, trimethylbenzene (TMB) as pore expander, tetraethyl orthosilicate (TEOS) as organosilicon source, water as solvent, and hydrochloric acid to adjust the pH value of the system to synthesize ordered silicon-based mesoporous Material SBA-15. The molar ratio of each material in the reaction is: 1TEOS: 0.017P123: 0.6TMB: 5.85HCl: 165H 2 O. Under magnetic stirring, add the silicon source into the template agent acidic aqueous solution, stir at room temperature for 2 hours, and then statically crystallize at 35°C for 24 hours; react in an autoclave at 100°C for 24 hours, filter, and wash with deionized water until Neutral, after drying at room temperature, calcined at 550°C for 4 hours to obtain SBA-15 carrier. The carrier has a most probable pore diameter of 9.8nm and a specific surface area of ​​977m 2 / g.

[0021] Add 5 g of the mesoporous material prepared by the above method to 3 mg / ml alkaline protease, shake at room temperature for 1...

Embodiment 2

[0023] Weigh 0.50g of casein, accurate to 0.01g, moisten it with a small amount of 0.5mol / l sodium hydroxide solution, add about 800ml of buffer solution with an appropriate pH=7.5, stir while heating in a boiling water bath until completely dissolved, After cooling, transfer to a 1000ml volumetric flask, dilute with buffer solution to volume.

Embodiment 3

[0025] Dissolve casein at a concentration of 0.5 mg / ml in a phosphate buffer solution with a pH of 8.0, flow through the reactor prepared in Example 1 above at a rate of 1000 ml / h at 50° C., collect the effluent, and concentrate , spray-dried protein peptides. The obtained protein peptide and the protein peptide hydrolyzed by free alkaline protease were subjected to HPLC analysis, and the results are shown in the accompanying drawings.

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Abstract

The invention is a method for preparing protein polypeptide by continuously hydrolyzing protein with an immobilized enzyme. The process comprises the steps of synthesizing an inorganic carrier for immobilizing protease; immobilizing the protease on the inorganic carrier; preparing an enzymatic hydrolysis reactor which can conduct continuous reaction; and subjecting a protein solution to enzymatic hydrolysis. The method has the advantages of simple process, high operability, mild reaction conditions, recyclable immobilized protease and low economic cost.

Description

technical field [0001] The invention relates to a method for producing protein peptides by enzymatically hydrolyzing proteins with proteases, in particular to a method for producing protein peptides using immobilized proteases. Background technique [0002] Protein peptide is a kind of biologically active substance, which is a mixed oligopeptide with certain biological functions obtained by proteolysis of protein or microbial fermentation. From a chemical point of view, protein peptide is a general term for different peptides ranging from dipeptides to complex linear and circular structures composed of 20 natural amino acids in proteins in different compositions and arrangements. Biological tests have shown that most of the protein ingested by the body is absorbed in the form of low-molecular-weight peptides after the action of enzymes in the digestive tract, and a small proportion is absorbed in the form of free amino acids. Further experiments revealed that low-molecular-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/00
Inventor 董颖超齐涛李军国李俊王丽娜
Owner FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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