Method for extracting pig blood antibiotic peptide
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A technology of antibacterial peptides and pig blood, applied in antibacterial drugs, medical raw materials derived from mammals, fermentation, etc., can solve the problems of extracting antibacterial peptides, and achieve the effect of simple equipment, convenient operation and low price
Active Publication Date: 2008-08-27
CHINA AGRI UNIV
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[0024] 1. Collect 1L of fresh pig blood from the slaughterhouse, then quickly add trisodium citrate to prevent coagulation according to the amount of 6g / L blood, and then filter it with gauze to remove the fibrous substances in it, and freeze and thaw the filtrate repeatedly to dissolve Blood (freeze-thawed 1-3 times between -20°C and 4°C), freeze-dried, and made into pig blood powder. The purpose of this step is to disrupt the porcine blood cells, releasing hemoglobin.
[0025] Pig blood meal also can adopt commercially available finished product pig blood meal.
[0026] 2, the pH that 2g pig blood meal is dissolved into 200ml is 7.0, and concentration is the disodium hydrogen phosphate-citric acid buffer solution (Lv Shiming, Tan Aijuan of 0.05mmol / L. Papain is to the hydrolysis of pig blood meal protein. Guizhou Agricultural Science, 2001 , 29(4):6~7), then add 20mg of papain (blood powder: ...
Embodiment 2
[0033] Embodiment 2, antibacterial and antiviral tests of porcine blood antimicrobial peptides
[0034] 1. Antibacterial activity experiment of pig blood antimicrobial peptide
[0035] The antimicrobial activity of the porcine blood antibacterial peptide extracted from pig blood in Example 1 was detected by using the agarose dispersion method. The tested bacteria are: Pasteurella multocida, natural isolates of chicken pathogenic Salmonella, Escherichia coli O 88 , Staphylococcus aureus ACTT25923 strain, Pseudomonas aeruginosa ACTT27853 strain.
[0036] The results of the determination of the antibacterial activity of porcine antimicrobial peptides showed that porcine antimicrobial peptides were effective against Pasteurella multocida, chicken pathogenic Salmonella, Escherichia coli O 88 , Staphylococcus aureus ACTT25923 strain, and Pseudomonas aeruginosa ACTT27853 strain all had obvious inhibitory effects, and the specific results are shown in Table 1.
[0037] Table 1 Anti...
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Abstract
The invention discloses a method for extracting porcine blood antibacterial peptide, which comprises following steps: firstly, dissolving porcine blood powder in buffer solution, then, adding papain into mixture to carry out enzymatic hydrolysis, adjusting the pH value of enzymolysis liquid to be 3.5-4.5, secondly, adding chloroform and sodium bisulfite into the enzymolysis liquid to decolorize, and then adjusting the pH value of the sodium bisulfite which is decolorized to be 6.0-7.0 and to form crude extracts of the porcine blood antibacterial peptide. Aiming at obtaining finished goods of the porcine blood antibacterial peptide, following purification steps are carried out to the crude extracts: firstly, freezing and drying the crude extracts of the porcine blood antibacterial peptide, enabling the crude extracts to pass through a gel-type cation exchange column, eluting the exchange column, collecting eluent, and secondly, freezing and drying the eluent in vacuum to form porcine blood antibacterial peptide. The method of the invention provides a new approach and a new method for producing antibacterial peptide. The method of the invention has the characteristics of simple operation, high separation extraction rate and low cost, and is suitable for separating and purifying the antibacterial peptide from porcine blood with comparatively large amount.
Description
technical field [0001] The invention relates to a method for extracting antimicrobial peptides, in particular to a method for extracting porcine blood antimicrobial peptides. Background technique [0002] At present, due to the extensive abuse of antibiotics, the problem of resistance to traditional antibiotics has attracted more and more attention. It has become an urgent task to seek a new type of antibacterial drug without residue and drug resistance. Antimicrobial peptides have gradually attracted people's interest and attention because of their high-efficiency antibacterial effects and broad and stable biological activities. Antimicrobial peptides have broad-spectrum and high-efficiency antimicrobial activity, and can also promote animal growth and induce animal acquired immunity. Moreover, its unique antibacterial mechanism will not cause microorganisms to develop drug resistance, and there is no residue in the body, so it has good application and development prospect...
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