Extractive fermentation technique or in situ product recovery process is a novel technique to segregate the desired product simultaneously in a fermentation process. For economic and high yield production of pullulan, Extractive fermentation process was applied fermentation process of A. pullulans. Aqueous Two Phase system (ATPS) systems were designed with various molecular mass of PEG (400, 600, 4000 and 6000) and dextran or mono/bi-sodium phosphate salts. Systems with short Tie Line length (TLL) 6.7 and 7.5% w/w for PEG-Salt and PEG-dextran respectively were chosen. Volume ratio for all the systems was kept constant at 1.0 and pH 7.0 for PEG-dextran and PEG-NaH2PO4 was maintained, whereas pH 9.0 was kept for PEG-Na2HPO4. A. pullulans, was found to be viable with PEG-NaH2PO4 and PEG-dextran systems. The biomass partitioned in the PEG rich top phase and the exopolysaccharide pullulan shown affinity towards the bottom phase. A maximum yield (36.47 g/L) was found with PEG 4000-Dextran 500 system of extractive fermentation process. The proposed process aptly integrates upstream and downstream process for continuous production and recovery of pullulan from the biomass, thus reducing the time quotient of the whole process.
Pullulan production from Aureobasidium pullulans was explored to increase yield. Non-linear hybrid mathematical tools for optimization of process variables as well as the pullulan yield were analyzed. The one variable at a time (OVAT) approach was used to optimize the maximum pullulan yield of 35.16 ± 0.29 g/L. The tools predicted maximum pullulan yields of 39.4918 g/L (genetic algorithm coupled with artificial neural network (GA–ANN)) and 36.0788 g/L (GA coupled with adaptive network based fuzzy inference system (GA–ANFIS)). The best regression value (0.94799) of the Levenberg–Marquardt (LM) algorithm for ANN and the epoch error (6.1055 × 10−5) for GA–ANFIS point towards prediction precision and potentiality of data training models. The process parameters provided by both the tools corresponding to their predicted yield were revalidated by experiments. Among the two of them GA–ANFIS results were replicated with 98.82% accuracy. Thus GA–ANFIS predicted an optimum pullulan yield of 36.0788 g/L with a substrate concentration of 49.94 g/L, incubation period of 182.39 h, temperature of 27.41 °C, pH of 6.99, and agitation speed of 190.08 rpm.
Pullulan is a commercially important Exopolysaccharide (EPS) with wide‐spread applications which is produced by Aureobasidium pullulans. The alternative α (1 4) & α (1 6) configuration in pullulan provides it the specific structural and conformational properties. Pullulan is currently being exploited in food, health care, pharmacy, lithography, cosmetics. The fermented broth is processed by organic solvent precipitation for isolation and purification of pullulan. In this study, we have tried to analyze the potential of aqueous two phase system as an alternate technique to extract pullulan from fermented broth. Including this viability of ATPS was also compared with conventional organic solvent precipitation system in terms of cost and time. It was found that ATPS process produced a higher yield of pullulan (80.56%) than organic solvent precipitation method (71.6%). ATPS was also found more economical and less time consuming method.
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