A fed-batch strategy was established based on the maximum substrate uptake rate (MSUR) of Pseudomonas aeruginosa USM-AR2 grown in diesel to produce rhamnolipid. This strategy matches the substrate feed rates with the substrate demand based on the real-time measurements of dissolved oxygen (DO). The MSUR was estimated by determining the time required for consumption of a known amount of diesel. The MSUR trend paralleled the biomass profile of Ps. aeruginosa USM-AR2, where the MSUR increased throughout the exponential phase indicating active substrate utilization and then decreased when cells entered stationary phase. Rhamnolipid yield on diesel was enhanced from 0·047 (g/g) in batch to 0·110 (g/g) in pulse-pause fed-batch and 0·123 (g/g) in MSUR fed-batch. Rhamnolipid yield on biomass was also improved from 0·421 (g/g) in batch, 3·098 (g/g) in pulse-pause fed-batch to 3·471 (g/g) using MSUR-based strategy. Volumetric productivity increased from 0·029 g l(-1) h(-1) in batch, 0·054 g l(-1) h(-1) in pulse-pause fed-batch to 0·076 g l(-1) h(-1) in MSUR fed-batch.
A biosurfactant-producing and hydrocarbon-utilizing bacterium, Pseudomonas aeruginosa USM-AR2, was used to assist conventional distillation. Batch cultivation in a bioreactor gave a biomass of 9.4 g L(-1) and rhamnolipid concentration of 2.4 g L(-1) achieved after 72 h. Biosurfactant activity (rhamnolipid) was detected by the orcinol assay, emulsification index and drop collapse test. Pretreatment of crude oil TK-1 and AG-2 with a culture of P. aeruginosa USM-AR2 that contains rhamnolipid was proven to facilitate the distillation process by reducing the duration without reducing the quality of petroleum distillate. It showed a potential in reducing the duration of the distillation process, with at least 2- to 3-fold decreases in distillation time. This is supported by GC-MS analysis of the distillate where there was no difference between compounds detected in distillate obtained from treated or untreated crude oil. Calorimetric tests showed the calorie value of the distillate remained the same with or without treatment. These two factors confirmed that the quality of the distillate was not compromised and the incubation process by the microbial culture did not over-degrade the oil. The rhamnolipid produced by this culture was the main factor that enhanced the distillation performance, which is related to the emulsification of hydrocarbon chains in the crude oil. This biotreatment may play an important role to improve the existing conventional refinery and distillation process. Reducing the distillation times by pretreating the crude oil with a natural biosynthetic product translates to energy and cost savings in producing petroleum products.
In this research we assess the feasibility of using palm oil agricultural refinery waste as a carbon source for the production of rhamnolipid biosurfactant through fermentation. The production and characterization of rhamnolipid produced by Pseudomonas aeruginosa PAO1 grown on palm fatty acid distillate (PFAD) under batch fermentation were investigated. Results show that P. aeruginosa PAO1 can grow and produce 0.43gL(-1) of rhamnolipid using PFAD as the sole carbon source. Identification of the biosurfactant product using mass spectrometry confirmed the presence of monorhamnolipid and dirhamnolipid. The rhamnolipid produced from PFAD were able to reduce surface tension to 29mNm(-1) with a critical micelle concentration (CMC) 420mgL(-1) and emulsify kerosene and sunflower oil, with an emulsion index up to 30%. Results demonstrate that PFAD could be used as a low-cost substrate for rhamnolipid production, utilizing and transforming it into a value added biosurfactant product.