Isolation, Screening and Identification of Biosurfactant Producing Strain <i>Nocardiopsis dassonvillei</i> var B2 From Oil Contaminated Soil

Petroleum and other oil manufacturing industries contribute to environmental pollution by releasing hazardous hydrocarbons. Biosurfactants offer a sustainable solution for mitigating oil pollution through emulsification processes, safeguarding agricultural soils, aquatic ecosystems, and human health. This study focuses on isolating, screening, and identifying actinomycetes producing biosurfactant from oil-polluted soil in the naval dockyard of Visakhapatnam. Biosurfactant production was successfully achieved utilizing Kim's medium, which was supplemented with olive oil serving as the carbon source. The evaluation involved preliminary identification tests, including oil displacement, Parafilm-M, and lipase activity assays, using sodium lauryl sulfate as the standard reference. Surface tension and emulsification index measurements were conducted, and the chemical composition of glycolipids and phospholipids was elucidated using phenol-sulfuric acid and phosphate assays. Glycolipids, specifically identified as rhamnolipids, were confirmed via cetyltrimethylammonium bromide (CTAB) testing and quantitatively analyzed using the orcinol method. The cell-free broth exhibited antagonistic activity against Gram-positive and negative bacilli.16S rRNA sequencing-based phylogenetic analysis was carried out by the NCIM, Pune, with the gene sequence being deposited in GenBank. Further characterization of isolate B2 included scanning electron microscopy (SEM) analysis, as well as physiological and biochemical assays. This study highlights the ability of Nocardiopsis dassonvillei var. B2, isolated from oil-polluted soil, to produce biosurfactants, specifically glycolipids identified as rhamnolipids. Our findings represent the first reported instance of biosurfactant production from isolate B2 originating from the naval dockyard in Visakhapatnam, Andhra Pradesh, India.