Characterization of Bioactive Secondary Metabolites Produced by Streptococcus pyogenes Using Gas Chromatography-Mass Spectrometry (GC-MS) Evaluation of Its Antibacterial Activity

Abstract

Background: Metabolites are small molecules participating in metabolic reactions, which are necessary for cellular function, maintenance and growth. Typically, metabolites range from 50 to 1500 Da, while their concentrations span several orders of magnitude. The metabolome is highly dynamic, time-dependent, and metabolites are sensitive to many environmental conditions. Aims and Objectives: The purpose of this laboratory study was to study the truly biologically active chemical compounds produced by Streptococcus pyogenes and to evaluate the antibacterial bioactivity of these products. Method: Streptococcus pyogenes were isolated from pediatric patients with streptococcal pharyngitis, all swabs collected at Babylon Hospital for Women and Children, Samples were incubated in stationary culture at 37°C for 24 h, after which headspace collections were made. All bacterial species were grown on identical media with identical glassware. Concentration and analysis of volatile compounds : Volatile metabolites were concentrated and separated and analyzed via GC-MS. In this research, (GC-MS) techniques were used to investigate the biochemical components that are commonly referred to as bioactive substances. Furthermore, the ethanolic extract of Streptococcus pyogenes was tested in an experimental laboratory to determine whether it actually has effective antibacterial properties. Results: Using GC-MS analysis on Streptococcus pyogenes, the presence of the following bioactive components was experimentaly identified: Carvacrol, 1-(5(Methyl-2-furanyl)-1-buten-3-one, beta Sesquiphellandrene, Pentasiloxane, dodecamethyl, Pyrazine, 2,5-dimethyl-, Benzene, 2-ethyl-1,4-dimethyl, tumerone, AR- tumerone, ALPHA- tumerone, 6-Aza-5,7,12,14-tetrathiapentacene, β-HIMACHALENOXIDE, 3-Ethyl-o-xylene, Trimethylphenylsilane, 1,3-Hexadiene, 2,5-dimethyl, o-Mercaptoaniline, Benzonitrile, 4-amino, Cyclononasiloxane, octadecamethyl, 3-Decen-5-one, Cyclononasiloxane, octadecamethyl, Methyl linoleate, Gentisic acid, Shogaol, Isooctyl phthalate, 2,6,10,-Pentamethyl-2,6, 18-eicosapentaene, Heptasiloxane, hexadecamethyl, N-Methyl-1-adamantaneacetamide. It was investigated whether the secondary metabolites produced by Streptococcus pyogenes have antibacterial properties against three dangerous pathogens. In the current research, the biological activity of the ethanolic extract of Streptococcus pyogenes, as well as the conventional antibiotics Rifambin and Cefotoxime, against three different pathogens were investigated. Escherichia coli (14.09±0.18, 20.17±0.25, and 17.37±0.23), Proteus mirabilis (12.91±0.16, 19.47±0.23, and 15.11±0.19), and Staphylococcus aureus (11.01±0.14, 20.07±0.25, and 17.00±0.21). Streptococcus pyogenes metabolites were shown to show remarkable activity against Escherichia coli, with a mean value of 17.37±0.23.