title: A simple, robust enzymatic-based high-throughput screening method for antimicrobial peptides discovery against Escherichia coli.
abstract: The indiscriminate usage of antibiotics has created a major problem in the form of antibiotic resistance. Even though new antimicrobial drug discovery programs have been in place from the last two decades, still we are unsuccessful in identifying novel molecules that have a potential to become new therapeutic agents for the treatment of microbial infections. A major problem in most screening studies is the requirement of high-throughput techniques. Given this, we present here an enzyme-based robust method for screening antimicrobial agent's active against Escherichia coli. This method is based upon the ability of the intracellular innate enzyme to cleave o-nitrophenyl β-d-galactopyranoside (non-chromogenic) to o-nitrophenolate (ONP) (chromogenic) upon the membrane damage or disruption. In comparison with the other currently available methods, we believe that our method provides an opportunity for real-time monitoring of the antimicrobial agents action by measuring the ONP generation in a user-friendly manner. Even though this method can be applied to other strain, our experience shows that one has to be careful especially when the pigments or metabolites present in the bacteria have the same wavelength absorbance.
authors.affiliation: SRM Research Institute, SRM University, Chennai, Tamil Nadu, India.
authors.forename: Muthukumaresan Kuppuswamy
authors.initials: MK
authors.lastname: Thirumalai
authors.forename: Arpita
authors.initials: A
authors.lastname: Roy
authors.forename: Suma
authors.initials: S
authors.lastname: Sanikommu
authors.forename: Jesu
authors.initials: J
authors.lastname: Arockiaraj
authors.forename: Mukesh
authors.initials: M
authors.lastname: Pasupuleti
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D023181:Antimicrobial Cationic Peptides / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D055808:Drug Discovery; D004926:Escherichia coli / Q000187:drug effects* / Q000254:growth & development; D057166:High-Throughput Screening Assays; D008826:Microbial Sensitivity Tests; D013329:Structure-Activity Relationship; D001616:beta-Galactosidase / Q000737:chemistry / Q000378:metabolism
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D001616:beta-Galactosidase
keywords: anti-infective agents; antibiotics; antimicrobial peptides; antimicrobial resistance; o-Nitrophenolate; o-Nitrophenyl β-d-galactopyranoside; β-galactosidase