Valorization of waste engine oil to mono- and di-rhamnolipid in a sustainable approach to circular bioeconomy.

This study aims to valorize waste engine oil (WEO) for synthesizing economically viable biosurfactants (rhamnolipids) to strengthen the circular bioeconomy concept. It specifically focuses on investigating the influence of key bioprocess parameters, viz. agitation and aeration rates, on enhancing rhamnolipid yield in a fed-batch fermentation mode. The methodology involves conducting experiments in a stirred tank bioreactor (3 L) using Pseudomonas aeruginosa gi |KP 163922| as the test organism. Central composite design and response surface methodology (CCD-RSM) are employed to design the experiments and analyze the effects of agitation and aeration rates on various parameters, including dry cell biomass (DCBM), surface tension, tensoactivity, and rhamnolipid yield. It is also essential to determine the mechanistic pathway of biosurfactant production followed by the strain using complex hydrophobic substrates such as WEO. The study reveals that optimal agitation and aeration rates of 200 rpm and 1 Lpm result in the highest biosurfactant yield of 29.76 g/L with minimal surface tension (28 mN/m). Biosurfactant characterization using FTIR, 1H NMR, and UPLC-MS/MS confirm the presence of dominant molecular ion peaks m/z 543.9 and 675.1. This suggests that the biosurfactant is a mixture of mono- and di-rhamnolipids (RhaC10C10, RhaRhaC10C12:1, RhaRhaC12:1C10). The findings present a sustainable approach for biosurfactant production in a fed-batch bioreactor. This research opens the possibility of exploring the use of pilot or large-scale bioreactors for biosurfactant production in future investigations.

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

ABSTRACTEnvironmental 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.

Production, characterization, and kinetic modeling of biosurfactant synthesis by Pseudomonas aeruginosa gi |KP 163922|: a mechanism perspective.

Kinetic studies and modeling of production parameters are essential for developing economical biosurfactant production processes. This study will provide a perspective on mechanistic reaction pathways to metabolize Waste Engine Oil (WEO). The results will provide relevant information on (i) WEO concentration above which growth inhibition occurs, (ii) chemical changes in WEO during biodegradation, and (iii) understanding of growth kinetics for the strain utilizing complex substrates. Laboratory scale experiments were conducted to study the kinetics and biodegradation potential of the strain Pseudomonas aeruginosa gi |KP 163922| over a range (0.5-8% (v/v)) of initial WEO concentration for 168 h. The kinetic models, such as Monod, Powell, Edward, Luong, and Haldane, were evaluated by fitting the experimental results in respective model equations. An unprecedented characterization of the substrate before and after degradation is presented, along with biosurfactant characterization. The secretion of biosurfactant during the growth, validated by surface tension reduction (72.07 ± 1.14 to 29.32 ± 1.08 mN/m), facilitated the biodegradation of WEO to less harmful components. The strain showed an increase in maximum specific growth rate (µmax) from 0.0185 to 0.1415 h-1 upto 49.92 mg/L WEO concentration. Maximum WEO degradation was found to be ~ 94% gravimetrically. The Luong model (adj. R2 = 0.97) adapted the experimental data using a non-linear regression method. Biochemical, 1H NMR, and FTIR analysis of the produced biosurfactant revealed a mixture of mono- and di- rhamnolipid. The degradation compounds in WEO were identified using FTIR, 1H NMR, and GC-MS analysis to deduce the mechanism.