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Polypeptide compound with broad-spectrum bactericidal activity on gram-positive bacteria

A polypeptide complex, Gram-positive technology, applied in the field of biomedicine, can solve the problems of knowing that the polypeptide complex is killed, and the polypeptide complex does not appear.

Pending Publication Date: 2021-11-30
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present inventors have studied a polypeptide complex with broad-spectrum bactericidal activity against Gram-positive bacteria, which belongs to antimicrobial peptides, which can kill a variety of bacteria including drug-resistant pathogenic bacteria, and the polypeptide complex has not appeared in known In the literature, it is difficult for those of ordinary skill in the art to know the beneficial effect of the polypeptide complex for killing bacteria through the existing literature

Method used

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  • Polypeptide compound with broad-spectrum bactericidal activity on gram-positive bacteria
  • Polypeptide compound with broad-spectrum bactericidal activity on gram-positive bacteria
  • Polypeptide compound with broad-spectrum bactericidal activity on gram-positive bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] This example provides an extraction, isolation and identification test of a polypeptide complex with broad-spectrum bactericidal activity against Gram-positive bacteria.

[0021] (1) Extraction of polypeptide complex

[0022] Centrifuge the fermentation broth of standard sensitive Staphylococcus aureus (ATCC 29213) at 4800r / min for 20min, take 50mL of the supernatant and place it in a 100mL beaker, and slowly add ammonium sulfate to it successively until the saturation of ammonium sulfate is 80 %, placed in a refrigerator at 4°C overnight, centrifuged at 12000r / min at 4°C for 20min with a high-speed centrifuge, dissolved in 20mmol / LTris-HCl buffer solution (pH7.5) and transferred to a treated dialysis bag, and placed in 20mmol / LTris-HCl buffer dialysis, with BaCl 2 After no precipitation was detected, the liquid was taken out and freeze-dried to measure its activity.

[0023] (2) Separation and purification of polypeptide complexes

[0024] Collect the above liquid a...

Embodiment 2

[0028] This example provides a bactericidal test of a polypeptide complex with broad-spectrum bactericidal activity against Gram-positive bacteria.

[0029] The strains used in the test were standard-sensitive Staphylococcus aureus (ATCC 29213), which produced the polypeptide complex with broad-spectrum bactericidal activity against Gram-positive bacteria in its culture solution. The strains tested included S.aureus 3521, S.aureus 1611, S.aureus Mu50, S.aureus Mu50 ΔrecA, S.heamolyticus NW19, S.heamolyticus 5211, S.aureus 1511, S.aureus 3821, S.aureus E48, S.agnetis 1932, S. epidermidis 2311, S. warneri 3511, Bacilus licheniformis CK1, Bacillus subtilis 1A747, E. coli K88, E. coli DH5a. In the test, standard sensitive Staphylococcus aureus (ATCC 29213) strains and strains to be tested were picked and inoculated into MH broth in sterilized test tubes, cultured overnight at 37°C and 250r / min. Transfer 30 L of the cultured bacterial liquid to 3 mL of fresh MH broth liquid medium...

Embodiment 3

[0035] This example provides a stability test of a polypeptide complex with broad-spectrum bactericidal activity against Gram-positive bacteria.

[0036] (1) Effect of concentration on the bactericidal activity of the polypeptide complex

[0037] Take 1.0 mL of polypeptide complexes with concentrations of 250, 125, 62.5, and 31.25 mg / mL, respectively, and distribute them into 4 centrifuge tubes, using Escherichia coli as the indicator bacteria. The indicator bacteria were inoculated in LB liquid medium and cultured at 37°C for 24 hours. Take 100 μL of the sample solution after filter sterilization, add 30 μL of LB medium and 70 μL of bacterial suspension (the concentration of bacterial suspension is 10 3 ~10 4 CFU / mL), mix well, incubate at 37°C for 1 hour on a shaker at 150 r / min, take 100 μL and pour it on a solid medium plate, culture overnight at 37°C, and count the number of colonies. Deionized water was used as the control group to calculate the antibacterial rate. T...

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Abstract

The invention discloses a polypeptide compound with broad-spectrum bactericidal activity on gram-positive bacteria. The polypeptide compound is formed by combining polypeptides with sequences shown in SEQ ID No.1 and SEQ ID No.2. The polypeptide compound with broad-spectrum bactericidal activity on gram-positive bacteria is derived from standard sensitive staphylococcus aureus (ATCC 29213). The polypeptide compound disclosed by the invention has broad-spectrum bactericidal activity on gram-positive bacteria and has a killing effect on methicillin-resistant bacteria and vancomycin-resistant bacteria which are difficultly killed by antibiotics.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a polypeptide complex with broad-spectrum bactericidal activity against Gram-positive bacteria. Background technique [0002] The extensive use of antibiotics has led to the evolution of antibiotic resistance in clinical pathogens, and the emergence of drug-resistant pathogens has greatly reduced the bactericidal effect and clinical expectations of antibiotics. In recent years, relevant research at home and abroad has found that not only some peptides extracted from organisms have biological activity, but also active peptides with special functions will be produced after protein hydrolysis. [0003] Antimicrobial peptides (AMPs) are a class of small molecular protein substances with antibacterial activity, which are widely distributed in various organisms and are key factors in the nonspecific immunity of invertebrates, vertebrates and plants. Antimicrobial peptides are considered to be...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/31A61P31/04
CPCC07K14/31A61P31/04
Inventor 刘丕龙李龙王利红赵丽丽严顺华
Owner NORTHWEST A & F UNIV
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