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Lactoferrin antimicrobial peptide, preparing method and uses thereof

A lactoferrin and antimicrobial peptide technology, applied in the field of lactoferrin antimicrobial peptide and its preparation, can solve problems such as lack

Inactive Publication Date: 2008-10-29
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, there is still a lack of methods for extracting lactoferrin antimicrobial peptides that can be widely used in practice, and the properties and uses of these lactoferrin antimicrobial peptides. Therefore, the applicant has carried out a large number of experimental studies and finally made the present invention

Method used

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  • Lactoferrin antimicrobial peptide, preparing method and uses thereof
  • Lactoferrin antimicrobial peptide, preparing method and uses thereof
  • Lactoferrin antimicrobial peptide, preparing method and uses thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0099] Weigh the lactoferrin sold by DMV Company under the trade name lactoferrin (Lactoferrin) with a purity greater than 90% and put it in an enzyme reactor, add deionized water and stir to obtain a concentration of 50mg / mL lactoferrin. Pretreatment at 40°C for 20 minutes, then adjust the pH of the solution to 2.5 with 1.0mol / L HCl, add 0.5% EC3.4.23.1 pepsin produced by Sigma in terms of lactoferrin weight, and then control the pH of the reaction system Constant, carry out enzymolysis at this temperature, use the TNBS method to measure the degree of hydrolysis until the degree of hydrolysis reaches 11%, after the reaction is completed, inactivate the enzyme at 100°C for 5 minutes, adjust the pH of the solution to 7.0, and proceed at 10000r / min Refrigerated and centrifuged for 20 min, the supernatant was concentrated. The bacteriostatic rate of the lactoferrin antimicrobial peptide hydrolyzate was 94.32% at 1 mg / mL.

[0100] The macroporous adsorption resin sold under the t...

Embodiment 2

[0105] This embodiment is implemented in the same manner as in Example 1, but the lactoferrin sold by DMV Company under the trade name lactoferrin (Lactoferrin) with a purity greater than 90% is put into an enzyme reactor, and deionized water is added to stir Uniformly prepare lactoferrin with a concentration of 10mg / mL, pretreat at 50°C for 10min, adjust the pH to 7.0 with 1.0mol / L NaOH, add 4% Novozyme 6.0S trypsin by weight of lactoferrin, during the reaction Keep the pH constant. After the reaction, inactivate the enzyme at 90°C for 8 minutes, adjust the pH to 7.0, refrigerate and centrifuge at 7000r / min for 30 minutes, concentrate the supernatant, and freeze-dry. thing. When the lactoferrin antimicrobial peptide hydrolyzate was at 1 mg / mL, the bacteriostatic rate was 67.39%.

[0106]Use DA201-B macroporous resin for static desalting, add lactoferrin antibacterial peptide, shake at 20°C at 150r / min for 12 hours, discard the supernatant, add deionized water, and wash until...

Embodiment 3

[0111] This embodiment is implemented in the same manner as in Example 1, but the lactoferrin sold by DMV Company under the trade name lactoferrin (Lactoferrin) with a purity greater than 90% is put into an enzyme reactor, and deionized water is added to stir Uniformly prepare lactoferrin with a concentration of 15mg / mL, pretreat at 50°C for 10 minutes, adjust the pH to 7.0 with 1.0mol / L NaOH, add 1.5% AS1398 neutral protease based on the weight of lactoferrin, and control the pH of the reaction system. After the reaction, inactivate the enzyme at 85°C for 10 minutes, adjust the pH to 7.0, and refrigerate and centrifuge at 9000 r / min for 20 minutes, and concentrate the obtained supernatant. Use DA201-D macroporous resin for static desalting, add lactoferrin antibacterial peptide at 160r / min, stir for 7 hours, discard the supernatant, add deionized water, wash until the conductivity is less than 15μs / cm, add 85 % ethanol for elution, the eluate was concentrated, and freeze-drie...

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Abstract

The invention relates to a lactoferrin antibacterial peptide, the preparation method and the application thereof. The method utilizes the lactoferrin and the enzyme preparation in bovine colostrum to prepare the lactoferrin antibacterial peptide zymolyte through enzymic hydrolysis, and subjects the zymolyte to macroporous absorption static or dynamic desalination, gel filtration chromatography, half-preparation high efficiency liquid chromatography and analysis-type high efficiency liquid chromatography for separation and purification to obtain a pure lactoferrin antibacterial peptide product. The product has high-efficient and broad-spectrum antibacterial capability, thereby having broad prospects for application in food, feed and medical fields.

Description

【Technical field】 [0001] The present invention relates to the technical field of bioactive peptides, in particular, the present invention relates to lactoferrin antibacterial peptides, their preparation methods and their uses. 【Background technique】 [0002] For more than 30 years, the research on new synthetic antibiotics has progressed slowly, and only a few reports on the chemical structures of new antibiotics have appeared. Moreover, these new antibiotics have a narrow antibacterial spectrum, limited application range, and high toxicity, which limits their widespread use. On the one hand, it seriously destroys the micro-ecological balance of the animal intestines and remains in the animals; on the other hand, it is easy to stimulate the production of drug-resistant strains, which leads to the increasing complexity of livestock and poultry diseases, and seriously affects the quality of livestock products and human health. Human beings' awareness of their own health and e...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K1/14C07K1/16C07K14/435A23L1/305A23L3/3526A23L2/44A23C3/08A23K1/16A61K38/17A61P31/00A23K20/147A23L33/18
Inventor 卢蓉蓉王新保
Owner JIANGNAN UNIV
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