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Cottonseed globulin positive charge antibacterial peptide and application thereof

A technology of antimicrobial peptides and globulins, which is applied in the fields of application, protein-containing food ingredients, peptides, etc., can solve the problems of unsatisfactory use requirements, low activity of natural antimicrobial peptides, and no cell selectivity, so as to promote healthy and sustainable development, inhibit Significant effect of bacterial activity

Pending Publication Date: 2021-12-31
CHENGUANG BIOTECH GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although they have significant differences in amino acid sequence and structure, most of them have two common features: (1) they are all cationic peptides, and the active structure is inducible amphiphilic polarity; (2) due to many natural antibacterial Peptides have low activity or no cell selectivity (that is, they are not only resistant to bacterial cells, but also toxic to mammalian cells), making these natural antimicrobial peptides often unable to meet people's use requirements

Method used

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  • Cottonseed globulin positive charge antibacterial peptide and application thereof
  • Cottonseed globulin positive charge antibacterial peptide and application thereof
  • Cottonseed globulin positive charge antibacterial peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The preparation of cottonseed globulin positively charged antimicrobial peptide comprises the following steps:

[0028] (1) Cottonseed protein was graded and extracted by Osborne method to prepare cottonseed globulin

[0029] Take 50g of defatted cottonseed protein and place it in a 1500mL large beaker, add 1000mL of distilled water, stir and extract at room temperature for 2 hours, then centrifuge at 6000r / min for 10min, take the lower layer of precipitate and use 1000mL of 1mol / L NaCl solution to stir and extract at room temperature 2h, then centrifuge at 6000r / min for 10min, filter the supernatant, and dialyze the obtained supernatant with a dialysis membrane with a molecular cut-off of 10000Da for 48h, change the water once every 1h, and put it in the dialysis bag after the dialysis is completed. The supernatant was vacuum freeze-dried to obtain cottonseed globulin.

[0030] (2) Alkaline protease is used to enzymatically hydrolyze cottonseed globulin

[0031] Add ...

Embodiment 2

[0060] Positively charged antimicrobial peptide F from cottonseed globulin AMP Antibacterial Activity Determination of

[0061] The antibacterial activity of positively charged antimicrobial peptides from cottonseed globulin was investigated by Oxford cup method, and three strains of Escherichia coli (ATCC25922), Staphylococcus aureus (ATCC25923) and Salmonella (ATCC14028) in logarithmic growth phase were diluted and evenly coated Spread it on a suitable plate so that the colonies can cover the entire plate, put an Oxford cup on the plate, and mix the antimicrobial peptide F AMP Dilute to different concentrations (0mg / ml, 0.1mg / ml, 0.2mg / ml, 0.4mg / ml, 0.8mg / ml, 1.6mg / ml, 3.2mg / ml, a total of 7 concentration gradients), take 200ul into the Oxford cup Placed in a 37°C incubator for 16 hours to observe the size of the inhibition zone.

[0062]

[0063] The results in the table above show that the maximum diameter of the inhibition zone to Salmonella is 6 mm, the maximum diam...

Embodiment 3

[0065] Effects of Environmental Factors on Cottonseed Globulin Positively Charged Antimicrobial Peptide F AMP The antibacterial activity of the

[0066] (1) Treatment of antimicrobial peptide F at different temperatures AMP

[0067] cottonseed globulin positively charged antimicrobial peptide F AMP Configured as antimicrobial peptide F with a concentration of 6.4mg / mL AMP aqueous solution, and placed at 5 different temperatures (4°C, 55°C, 80°C, 100°C and 120°C), and each temperature was maintained for 30 minutes.

[0068] (2) Different pH treatment of antimicrobial peptide F AMP

[0069] Acetate buffer solution of pH 3.0, 4.0 and 5.0, sodium phosphate buffer solution of pH 6.0 and 7.0, 0.1mol Tris-HCl buffer solution of pH 8.0 and pH 9.0 were used respectively, and the concentration was 6.4mg / mL antimicrobial peptide F AMP solution, adjust the pH to 7.0 after 20 min of each pH treatment.

[0070] (3) Treated antimicrobial peptide F AMP Bacteriostatic activity investi...

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Abstract

The invention relates to a cottonseed globulin positive charge antibacterial peptide and application thereof, and belongs to the technical field of antibacterial peptides, and the antibacterial peptide is an antibacterial peptide FAMP with an amino acid sequence as shown in SEQ ID NO.1. According to the invention, cottonseed protein is taken as a research object, extraction, enzymolysis, separation, purification and identification are carried out, the cottonseed globulin positive charge antibacterial peptide FAMP with a brand-new amino acid sequence is found, the antibacterial peptide FAMP has remarkable bacteriostatic activity on staphylococcus aureus, escherichia coli and salmonella, has no toxicity on normal red blood cells of animals, part of antibiotics can be replaced, and healthy and sustainable development of the breeding industry in China is promoted.

Description

technical field [0001] The invention relates to a positively charged antibacterial peptide of cottonseed globulin and an application thereof, belonging to the technical field of antibacterial peptides. Background technique [0002] The excessive use of antibiotics in the feed industry has led to bacterial resistance, causing many problems such as food safety and environmental pollution, and posing a serious threat to human health and safety. Therefore, exploring the addition of efficient, safe and green alternatives to antibiotics in feed has become a research hotspot. [0003] Antimicrobial peptide is a general term for a class of polypeptides with antibacterial activity. It has the characteristics of small molecular weight, strong heat resistance, broad antibacterial spectrum, low toxicity and no residual hazards. It has extensive research value in food, medicine, agriculture and feed industries. It is expected to become a substitute for chemical preservatives and farming...

Claims

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Application Information

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IPC IPC(8): C07K14/415C12P21/06C07K1/18C07K1/16C07K1/14A23L3/3526A23K20/147
CPCC07K14/415C12P21/06A23L3/3526A23K20/147A23V2002/00A23V2200/10A23V2250/55
Inventor 宋伟光陈运霞焦利卫曹通
Owner CHENGUANG BIOTECH GRP CO LTD
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