ABSTRACT
The biosensing of bacterial pathogens is of a high priority. Electrochemical biosensors are an important future tool for rapid bacteria detection. A monolayer of bacterial-binding peptides can serve as a recognition layer in such detection devices. Here, we explore the potential of random peptide mixtures (RPMs) composed of phenylalanine and lysine in random sequences and of controlled length, to form a monolayer that can be utilized for sensing. RPMs were found to assemble in a thin and diluted layer that attracts various bacteria. Faradaic electrochemical impedance spectroscopy was used with modified gold electrodes to measure the charge-transfer resistance (RCT) caused due to the binding of bacteria to RPMs. Pseudomonas aeruginosa was found to cause the most prominent increase in RCT compared to other model bacteria. We show that the combination of highly accessible antimicrobial RPMs and electrochemical analysis can be used to generate a new promising line of bacterial biosensors.
Subject(s)
Antimicrobial Peptides , Bacteria , Biosensing Techniques , Bacteria/isolation & purification , Biosensing Techniques/methods , Dielectric Spectroscopy/methods , Electrodes , Gold/chemistry , Peptides/chemistryABSTRACT
In the current study, we evaluated the antimicrobial activity of randomly-sequenced peptide mixtures (RPMs) bearing hydrophobic and cationic residues that were immobilized on beads. We showed that these beads exhibit high and broad bactericidal activity against various pathogenic bacteria while possessing minimal hemolytic activity.
Subject(s)
Anti-Bacterial Agents/pharmacology , Antimicrobial Cationic Peptides/pharmacology , Methicillin-Resistant Staphylococcus aureus/drug effects , Pseudomonas aeruginosa/drug effects , Anti-Bacterial Agents/chemistry , Antimicrobial Cationic Peptides/chemistry , Hydrophobic and Hydrophilic Interactions , Microbial Sensitivity TestsABSTRACT
The shelf life of pasteurized bovine milk is limited by microorganism activity as surviving bacteria continue to grow in the bovine milk, eventually causing milk spoilage. In the current study, we used matrix-assisted laser desorption ionization time of flight mass spectrometry to identify pasteurized bovine milk-associated mesophilic and psychrotrophic bacteria. We have recently designed random cationic peptide mixtures that possess strong antimicrobial and antibiofilm properties. These compounds are cheap and easy to synthesize and represent a new class of antimicrobial agents. Here, we show that the random peptide mixtures are able to efficiently eradicate the bacteria identified as associated with pasteurized bovine milk, and reduced significantly the growth of Bacillus subtilis in milk. We propose these antimicrobial peptides as potential candidates for integration in bioactive milk and food packaging to prevent bacterial growth and extend the shelf life of food.
Subject(s)
Anti-Bacterial Agents/pharmacology , Bacillus subtilis/drug effects , Milk/microbiology , Pasteurization , Peptides/pharmacology , Animals , Anti-Bacterial Agents/chemical synthesis , Anti-Bacterial Agents/chemistry , Bacillus subtilis/growth & development , Biofilms/drug effects , Cattle , Dose-Response Relationship, Drug , Microbial Sensitivity Tests , Peptides/chemical synthesis , Peptides/chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Structure-Activity RelationshipABSTRACT
Many types of crops are severely affected by at least one important bacterial disease. Chemical control of bacterial plant diseases in the field vastly relies on copper-based bactericides, yet with limited efficacy. In this study, we explored the potential of two random peptide mixture (RPM) models as novel crop protection agents. These unique peptide mixtures consist of random combination of l-phenylalanine and l- or d-lysine (FK-20 and FdK-20, respectively) along the 20 mer chain length of the peptides. Both RPMs displayed powerful bacteriostatic and bactericidal activities towards strains belonging to several plant pathogenic bacterial genera, for example, Xanthomonas, Clavibacter and Pseudomonas. In planta studies in the glasshouse revealed that RPMs significantly reduced disease severity of tomato and kohlrabi plants infected with Xanthomonas perforans and Xanthomonas campestris pv. campestris respectively. Moreover, RPM effects on reduction in disease severity were similar to those exerted by the commercial copper-based bactericide Kocide 2000 that was applied at a 12-fold higher concentration of the active compound relative to the RPM treatments. Importantly, the two tested RPM compounds had no toxic effect on survival of bees and Caco-2 mammalian cells. This study demonstrates the potential of these innovative RPMs to serve as crop protection agents against crop diseases caused by phytopathogenic bacteria.