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Sea-buckthorn seed polypeptide with inhibiting effect on intestinal pathogens as well as preparation method and application of sea-buckthorn seed polypeptide

A technology of antibacterial effect and seabuckthorn seeds, which is applied in the field of preparation of seabuckthorn polypeptides, and can solve problems such as interference with antibacterial effects

Active Publication Date: 2014-10-15
神池县曦晟源生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It mainly isolates small molecular salts, and some macromolecular substances that have no antibacterial effect remain in the seabuckthorn seed polypeptide solution, which interferes with the antibacterial effect

Method used

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  • Sea-buckthorn seed polypeptide with inhibiting effect on intestinal pathogens as well as preparation method and application of sea-buckthorn seed polypeptide
  • Sea-buckthorn seed polypeptide with inhibiting effect on intestinal pathogens as well as preparation method and application of sea-buckthorn seed polypeptide
  • Sea-buckthorn seed polypeptide with inhibiting effect on intestinal pathogens as well as preparation method and application of sea-buckthorn seed polypeptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Take 20g of pulverized defatted seabuckthorn seed dregs (30% protein content), add 300ml of distilled water, place it at 25°C for 8 hours, adjust the pH to 6.5, add 0.18g of Prote AX protease and hydrolyze it at 50°C for 3h; In a glass bottle, tighten the lid, use a 1000W household microwave oven to inactivate the enzyme for 5 minutes, centrifuge the enzymolysis solution at 4000r / min to get the supernatant, filter it with an ultrafiltration membrane with a cut-off pore size of 5000Da, and concentrate the filtrate to the peptide content Achieve 2% savings.

[0019] The above concentrated solution was diluted to 20mg / ml for antibacterial test, the bacteria used were Staphylococcus aureus and Salmonella, and the culture medium was nutrient agar medium. When the medium is boiled and cooled to 50-60°C, it is poured into a petri dish, and after solidification, Staphylococcus aureus and Salmonella are respectively inoculated on the surface of the medium. Make a small hole wit...

Embodiment 2

[0021] Take 50g of pulverized defatted seabuckthorn seed dregs (30% protein content), add 1000ml of distilled water, place it at 25°C for 10 hours, adjust the pH to 7.0, add 0.45g Prote AX protease, hydrolyze at 46°C for 4.5h, pack Put it into a sealed glass bottle, use a 1000W household microwave oven to inactivate the enzyme for 7 minutes, centrifuge the enzymolysis solution at 4000r / min to get the supernatant, filter it with an ultrafiltration membrane with a cut-off pore size of 5000Da, and concentrate the filtrate to the peptide content up to 16%.

[0022] Dilute the concentrated solution to 5, 10, 20, 40, 80, 160 mg / ml for antibacterial test, the bacteria used are Salmonella, Staphylococcus aureus and Pseudomonas aeruginosa, and the culture medium is nutrient agar medium. Cultivate and observe method with described in embodiment 1, obtain concentration inhibition curve ( image 3 ). It can be seen from the concentration inhibition curve that the optimal polypeptide inh...

Embodiment 3

[0024] Take 100g of pulverized defatted seabuckthorn seed dregs (30% protein content), add 1500ml of distilled water, place it at 25°C for 8 hours, adjust the pH to 7.0, add 0.63g of Prote AX protease, hydrolyze it at 55°C for 1.5h, put it into In a sealed glass bottle, use a 1200W household microwave oven to heat to inactivate the enzyme for 10 minutes, centrifuge the enzymolysis solution at 4000r / min to get the supernatant, filter it with an ultrafiltration membrane with a cut-off pore size of 5000Da, and take the filtrate and then filter it with a cut-off pore size of 4000Da The ultrafiltration membrane is used for ultrafiltration, and the concentrated solution is taken to separate short and medium molecular weight peptides of 4000 to 5000 Da. The filtrate is also used for ultrafiltration with an ultrafiltration membrane with a cut-off pore size of 2000 Da, and the filtrate is taken to obtain small molecular peptides below 2000 Da. Peptides of different molecular weights wer...

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Abstract

The invention provides a sea-buckthorn seed polypeptide solution with an inhibiting effect on intestinal pathogens as well as a preparation method and application of the sea-buckthorn seed polypeptide solution. The preparation steps are as follows: after sea-buckthorn seed residues are soaked, centrifugation is performed after ProteAX protease is added for hydrolysis, and the supernate is obtained, and is subjected to concentration and separation by an ultrafiltration membrane with the interception aperture of 5000 Da, so that the sea-buckthorn seed anti-bacteria polypeptide solution is obtained. The anti-bacteria culture experiment results show that the polypeptide solution within the concentration range of 10-20 mg / ml has the obvious inhibiting effect on salmonella, staphylococcus aureus, pseudomonas aeruginosa and other common intestinal pathogens. The sea-buckthorn seed polypeptide can be used as a feed additive.

Description

technical field [0001] The invention relates to the preparation of seabuckthorn polypeptides, in particular to the reuse of residues after oil extraction from seabuckthorn seeds, and relates to a seabuckthorn seed polypeptide solution with antibacterial effect and a preparation method and application thereof. Background technique [0002] Seabuckthorn seed dregs are the waste after seabuckthorn oil processing and production, usually only a small amount is used as feed, and most of them are discarded as garbage. However, the protein content in the seabuckthorn seed dregs after extracting seabuckthorn oil reaches 27-30%, and protein resources are wasted. At present, there is very little development and utilization of seabuckthorn seed dregs, mainly about the extraction of proanthocyanidins and flavonoids, but due to its high development cost, it has not been widely used. The research on seabuckthorn seed polypeptides includes "Preparation and Efficacy of Seabuckthorn Bioactiv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06A23K1/17A23K20/195
Inventor 王常青陈彤
Owner 神池县曦晟源生物科技有限公司
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