Aim: To identify indigenous bacteria responsible for producing biosurfactants in soil contaminated by oil and to investigate their potential uses in the fields of pharmaceuticals and environmental applications. Methodology: The microorganisms of oil-contaminated soil were isolated, screened, and identified using 16S rRNA gene sequencing. Bacillus amyloliquefaciens was utilized to produce biosurfactants, and were characterized by UV, TLC, FTIR, HPTLC, and GCMS analysis. Antibacterial activity, plant growth promotion, biocompatibility, and wound healing activity of surfactant were performed. Results: Six bacterial cultures (B1 to B6) were isolated from the soil samples collected from various locations in Kerala and Tamil Nadu. Among the isolates screened for biosurfactant production, Bacillus amyloliquefaciens exhibited the highest capacity for biosurfactant production, confirmed through 16S rRNA gene sequencing. The biosurfactant produced was identified as surfactin of lipopeptide nature, characterized by various analytical techniques: UV absorption at 400 nm, TLC and HPTLC (Rf value of 0.58), FTIR (prominent peak at 3819 cm-1 corresponding to -OH groups), GCMS with a maximum retention time of 12.03. Surfactin effectively inhibited pathogenic bacteria, enhanced plant growth, was biocompatible, and enhanced the rate of cell migration towards wound scratch. Interpretation: The current study revealed that Bacillus amyloliquefaciens, isolated from oil-contaminated soil, is capable of producing biosurfactant surfactin. Additionally, it unveiled a broad spectrum of applications for surfactin, encompassing wound healing, biocompatibility, antimicrobial activity, and promotion of plant growth. Key words: Antibacterial activity, Biocompatibility, Bacillus amyloliquefaciens, Biosurfactant, Wound healing
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