Rhamnolipid production by Pseudomonas aeruginosa (SSL-4) on waste engine oil (WEO): Taguchi optimization, soil remediation, and phytotoxicity investigation

Abstract

Environmental concerns and rising biosurfactant demand emphasize the need for this study. The objective is to maximize rhamnolipid-biosurfactant production by Pseudomonas aeruginosa (SSL-4) utilizing waste engine oil (WEO) as the sole substrate for use in soil bioremediation and commercial production. Using an L16 Taguchi orthogonal array, a signal-to-noise ratio, and an analysis of variance (ANOVA), the effects of environmental (pH, incubation temperature) and dietary parameters (carbon source concentration, carbon/nitrogen (C/N) and carbon/phosphorus (C/P) ratio) are examined. Variations of the following parameters were made within a carefully selected range: incubation temperature of 25–40℃, pH range of 5–11, WEO concentration of 1–7% (v/v), and C/N and C/P ratios of 10–40. Response variables in this batch study include surface tension reduction (mN/m), dry cell biomass (DCBM) (g/L), and rhamnolipids yield based on substrate consumption, YP/S (g/g). Rhamnolipid was synthesized under optimal conditions, providing a yield of 21.42 g/g. The oil recovery of 74.05 ± 1.481% was achieved from oil-contaminated soil at a CMC of ∼70 mg/L. FTIR, 1H NMR, and UPLC-MS techniques were utilized for the characterization of rhamnolipids, and AAS for determining heavy metals concentration in WEO and residual waste engine oil (RWEO). The Germination Index (GI) of ∼82.55% indicated no phytotoxicity associated with synthesized rhamnolipid.