Biosurfactants are surface-active compounds that have many benefits. This research aims to isolate and identify biosurfactants produced by B. subtilis 3KP when cultured on a molasses substrate. The isolation and purification procedure included gradual ammonium sulfate precipitation, ion exchange and adsorption chromatography. Identification and characterization using high-pressure liquid chromatography, gas chromatography–mass spectrometry, amino acid analyzer, UV–V is spectroscopy, and infrared spectroscopy confirmed that B. subtilis 3KP biosurfactant is a lipopeptide. The biosurfactant produced by B. subtilis 3KP was determined to be a surfactin isoform, characterized by a 16-carbon fatty acid chain. The purified biosurfactant successfully decreased the surface tension from 72 mN/m to 27 mN/m and reached the critical micelle concentration at a precise value of 20.01 mg/L. This biosurfactant product is stable at 30°C–65°C, pH 4–8, and salinity 0–4 M. Therefore, the presented isolate has tremendous potential to produce surfactin-type biosurfactants which can be developed and utilized for various industrial fields.

Keywords:
Biosurfactant, Bacillus subtilis, Lipopeptide, Characterization, Green Product
Subjects:
Biochemical Techniques & Resources, Biotechnology, Chemical Biology, Microbiology
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Copyright 2025 The Author(s)
2025
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