Proteomic Analysis of Bacterial Expression Profiles Following Exposure to Organic Solvent Flower Extract of Melastoma candidum D Don (Melastomataceae)
Purpose: To identify potential antibacterial protein targets following exposure to Melastoma candidum extract.
Methods: Plant extracts were prepared using sequential extraction method. Denaturing gel electrophoresis and MALDI TOF-TOF MS protein sequencing were used to identify differentialexpressed bacterial proteins. 96-well microplate method was used to determine the minimum inhibitory concentration (MIC) values. Thin layer chromatography (TLC) bio-autobiography and gaschromatography-mass spectrometry (GC-MS) were performed to determine the phytochemicals in the active fraction.
Results: Five differentially expressed bacterial proteins (four from Escherichia coli and one from Staphylococcus aureus), were identified via proteomic approach. Among the bacterial proteins identified, glutamate decarboxylase, elongation factor-Tu and α-hemolysin are especially noteworthy, as they are implicated in critical bacterial pathways pertaining to survival in acidic environment, protein translation and virulence, respectively. Additionally, we tested and reported the minimum inhibition concentrations of different M. candidum fractions and gas chromatography-mass spectrometry GC-MS analysis of the active fraction.
Conclusion: Glutamate decarboxylase, elongation factor-Tu and α-hemolysin represent potential antibacterial targets.
Keywords: Escherichia coli, Staphylococcus aureus, Melastoma candidum, Glutamate decarboxylase, Elongation factor-Tu, α-Hemolysin, Protein expression