Antimicrobial agent FAM-AMP with gelatinase digestion sequence for detection of S.aureus bacteria

A technology of FAM-AMP and gelatinase enzyme, which is applied in the field of antibacterial agents, can solve the problems of specificity and accuracy of damaging bacteria detection, and achieve the effect of being suitable for popularization, excellent antibacterial performance and high sensitivity

Pending Publication Date: 2020-01-10
CHANGZHOU UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, colorimetric assays, for example, are often susceptible to interference from environmental conditions (i.e., pH, ion concentration, and solvents), thus compromising the specificity and accuracy of bacterial detection.

Method used

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  • Antimicrobial agent FAM-AMP with gelatinase digestion sequence for detection of S.aureus bacteria
  • Antimicrobial agent FAM-AMP with gelatinase digestion sequence for detection of S.aureus bacteria
  • Antimicrobial agent FAM-AMP with gelatinase digestion sequence for detection of S.aureus bacteria

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Embodiment 1

[0032] 1. Synthesis of FAM-AMP

[0033] 1) Synthesis of AMP

[0034] AMP is based on 2-Chlorotrityl Chloride resin with Fmoc protecting group, obtained by solid-phase peptide synthesis. Specifically: weigh amino acid (chemically modified α-amino acid), HBTU, and HOBT respectively, which are equivalent to 5 times the molar amount of the resin, dissolve them in DMF, and add DIEA for coupling for 30 minutes. Then add 20% piperidine to react for 30 minutes to remove the Fmoc protecting group, repeat this step until the desired peptide chain is synthesized, and the synthesized peptide chain is purified by HPLC, and the purification effect is shown in the appendix figure 1 shown.

[0035] 2) 5-FAM fluorescein labeled AMP

[0036] Weigh 30 mg of resin (resin with peptide chains synthesized above), and dissolve 5-FAM, HOBT, EDC equivalent to 3 times the molar amount of the resin in 1 mL of DMF, add 10 μL of DIEA in the dark for overnight reaction, and finally cut the The labeled p...

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Abstract

The invention belongs to the technical field of antimicrobial agents, and discloses an antimicrobial agent FAM-AMP with gelatinase digestion sequence for detection of S.aureus bacteria. GKRWWKWWRR inAMP antimicrobial agent is antimicrobial peptide, PLGVRG is in gelatinase digestion sequence and can be recognized and cut off by the gelatinase secreted by S.aureus bacteria, and the PLGVRG and the gelatinase are coupled through peptide solid phase synthesis method to form new form of antimicrobial agent with gelatinase digestion points. Fluorescein FAM is marked into the antimicrobial agent AMPthrough coupling agents, and the marked FAM-AMP can further detect the concentration of the S.aureus through capillary electrophoresis. The antimicrobial agent FAM-AMP with the gelatinase digestion sequence for the detection of the S.aureus bacteria has the advantages that the concentration of the S.aureus bacteria can be detected, bacterium can be effectively killed, the biocompatibility is good,impact on normal physiological activities of body cells is small, toxicity is low and safety is high.

Description

technical field [0001] The invention belongs to the technical field of antibacterial agents, and specifically discloses an antibacterial agent FAM-AMP containing a gelatinase cleavage sequence for detection of S. aureus bacteria. Background technique [0002] Antibiotics have made outstanding contributions to anti-infection. However, the widespread use and abuse of antibiotics have led to the increasing resistance of many important human pathogens. new antibiotics. Antimicrobial peptides (AMPs) have incomparable advantages over traditional antibiotics. They have a unique antibacterial mechanism, rapid bactericidal effect and are not easy to cause bacterial drug resistance. They are a class of antibacterial drugs with great potential. Antimicrobial peptides are considered to have broad application prospects in the pharmaceutical industry because of their high antibacterial activity, wide antibacterial spectrum, many types, wide range of options, and the lack of resistance mu...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12Q1/14C12R1/445
CPCC07K7/06C12Q1/14C07K2319/60C07K2319/50G01N2333/31
Inventor 王建浩郭倩倩贾文静唐瑶王菲刘丽邱琳周舒文崔朋飞李玉亭赵俊泽
Owner CHANGZHOU UNIV
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