Optimization of biosurfactant has improved the value-chain, process development and cost of production associated with downstream synthesis. This study was designed to determine the optimal conditions for production of biosurfactant using hydrolyzed agroresidues under controlled conditions. Rhizobacterial isolate was obtained from Paspalum sp. growing on an aged crude oil impacted soil in Bodo, Rivers State, Nigeria. The bacterial isolates were identified using 16S rRNA molecular approach on a set of universal primers. One-Variable at a Time approach was applied for verification of pH, Carbon and nitrate sources respectively. Stat-ease Design-Expert version 12.0 was employed in the optimization of the variables while the operational conditions were fitted into a 20-run design matrix using α- level 2.0. Molecular identification confirmed the bacterial isolae to be Pseudomonas sp. with Accession number MH40927 with a gene molecular weight of 6.0kbp. Response for biomass, biosurfactant and Critical Miscelle Concentration (CMC) was observed to fit into a 2nd order Quadratic functions at p<0.05 with optimal conditions were pH = 7.0, Corn chaff = 2.0 g/L and Urea = 1.0 g/L. Biosurfactant = +51.98 + 6.79A + 4.30B‑5.45C + 0.2975AB + 0.4975AC + 1.63BC-6.46A² – 3.51B² – 6.94C². This study further identified a directly proportional relationship between biosurfactant production and operational variables which represents a cheaper and feasible production roadmap for biosynthesis.
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