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Preparation method for bacteriostatic seabuckthorn seed polypeptide

A technology of antibacterial effect and sea buckthorn seeds is applied in the field of preparation of plant polypeptides, which can solve the problems of astringent taste of polypeptides, large loss of polypeptides, low yield of polypeptides, etc., and achieve the effects of reducing loss of polypeptides, increasing the yield of hydrolysis, and good taste.

Inactive Publication Date: 2013-12-04
SHANXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it adopts the method of direct hydrolysis of protease, the process of preparing polypeptide by this method has a low yield of polypeptide, and the multi-step ultrafiltration membrane separation causes a large loss of polypeptide. with additives

Method used

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  • Preparation method for bacteriostatic seabuckthorn seed polypeptide
  • Preparation method for bacteriostatic seabuckthorn seed polypeptide
  • Preparation method for bacteriostatic seabuckthorn seed polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Take 20g of crushed seabuckthorn seed dregs in a beaker, add 200ml of distilled water, and place it overnight at room temperature of 25°C. Adjust the pH to 4.0, add 0.1 g of phytase and 0.1 g of tannase, and incubate in a water bath at 50° C. for 8 hours. Then heat to 90°C to inactivate the enzyme for 15 minutes, adjust the pH to 2.5 after cooling, add 0.33 g of acid protease to hydrolyze at 45°C for 7 hours, heat to 90°C to inactivate the enzyme for 15 minutes, adjust the pH to 7.0 after cooling, add 0.15 g of papain to the Hydrolyze at 55°C for 3h, heat to 90°C to inactivate the enzyme for 15min. Place the enzymolysis solution at 0-4°C for 8 hours. Centrifuge at a high speed of 12000r / min for 10min at 0-4°C, take the centrifuged supernatant and filter it with a nanofiltration membrane with a molecular weight cut-off of 500Da, and take the retentate of the nanofiltration for antibacterial test. The bacterium used in the antibacterial test was Salmonella typhimurium, ...

Embodiment 2

[0022] Take 20g of crushed seabuckthorn seed dregs in a beaker, add 300ml of distilled water and soak overnight. Adjust the pH to 4.5, add 0.2g phytase and 0.1g tannase, and keep it in a water bath at 45°C for 8h, stirring for 2-3min every 1h. Then heat to 90°C to inactivate the enzyme for 15 minutes, adjust the pH to 3.5 after cooling, add 0.5g acidic protease to hydrolyze at 50°C for 4 hours, heat to 90°C to inactivate the enzyme for 15 minutes, adjust the pH to 6.5 after cooling, add 0.1g papain to the Hydrolyze at 50°C for 5 hours, heat to 90°C to inactivate the enzyme for 15 minutes, then heat to 90°C to inactivate the enzyme for 15 minutes. Place the enzymolysis solution at 0-4°C for 10 hours. Centrifuge at a high speed of 10000r / min for 20min at 0-4°C, take the centrifuged supernatant and filter it with a nanofiltration membrane with a molecular weight cut-off of 500Da, and take the nanofiltration retentate for antibacterial test. The bacterium used in the antibacteri...

Embodiment 3

[0028] Take 20g of crushed seabuckthorn seed dregs in a beaker, add 400ml of distilled water and soak overnight. Adjust the pH to 5, add 0.2 g of phytase and 0.1 g of tannase, and incubate in a water bath at 38° C. for 14 hours. Then heat to 90°C to inactivate the enzyme for 15 minutes, adjust the pH to 4.0 after cooling, add 0.4g acidic protease to hydrolyze at 55°C for 3 hours, heat to 90°C to inactivate the enzyme for 15 minutes, adjust the pH to 7 after cooling, add 0.25g papain to the Hydrolyze at 52°C for 1.5h, then heat to 90°C to inactivate the enzyme for 15min. Place the enzymolysis solution at 0-4°C for 12 hours. Centrifuge at a high speed of 11000r / min for 15min at 0-4°C, take the centrifuged supernatant and filter it with a nanofiltration membrane with a molecular weight cut-off of 500Da, and take the nanofiltration retentate for antibacterial test. The bacterium used in the antibacterial test was Salmonella pullorum, and the culture medium was nutrient broth med...

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Abstract

The invention provides a preparation method for bacteriostatic seabuckthorn seed polypeptide. The preparation method comprises the following steps: soaking seabuckthorn seed slag for treatment, conducting hydrolytic treatment on the seabuckthorn seed slag with complex enzyme of tannase and phytase, hydrolyzing protein in zymolyte with the combination of acidic protease and papayotin, standing polypeptide hydrolysate at a low temperature for one night, removing macromolecular impurities through low-temperature high-speed centrifugation, and concentrating and drying centrifugation supernatant fluid to obtain the bacteriostatic seabuckthorn seed polypeptide. Extracorporeal bacteriostatic cultivation experiments show that the polypeptide has a favorable restraining effect for common bacteria such as colibacillus coli and salmonella. The polypeptide is good in taste, has no side effects, and can be added into food or fodder as a preservative agent.

Description

technical field [0001] The invention relates to the preparation of plant polypeptides, in particular to the reuse of the residue after degreasing seabuckthorn seeds, and is a method for preparing seabuckthorn seed polypeptides with antibacterial effect. Background technique [0002] Defatted seabuckthorn seed residue is the waste after seabuckthorn oil processing and production, which is usually used as animal feed or discarded. The defatted seabuckthorn seed residue contains about 30% protein, and the ratio of amino acids is good. At present, there is very little development and utilization of seabuckthorn seed dregs, only the development and utilization of extracting proanthocyanidins, but because its development cost is greater than that of common grape seeds, it has not been widely used. The research on seabuckthorn seed polypeptides includes "Preparation and Efficacy of Seabuckthorn Bioactive Peptides" published by Huang Peng, Suning and others. In this study, a polype...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06A23L3/3526A23K1/17A23K20/195
Inventor 王常青连伟帅
Owner SHANXI UNIV
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