Extractive Fermentation for Process integration and amplified pullulan production by A. pullulans in Aqueous Two Phase Systems

Parul Badhwar,Punit Kumar,Kashyap Kumar Dubey

doi:10.1038/s41598-018-37314-y

Parul Badhwar, Punit Kumar + Show 1 more

Open Access

https://doi.org/10.1038/s41598-018-37314-y

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Abstract

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.

Highlights

The development and designing of an efficient bioprocess is the most crucial step in any biotechnology industry
The prime objective of studying Aqueous Two Phase system (ATPS) here is to replace and integrate various unit operations involved in fermentative production of pullulan, with the single stage aqueous partitioning
Extractive fermentation or in situ product removal, is a novel process deriving from ATPS where the microbial system grows in one phase and the product of interest being extracellular in nature is excreted out and partitioned towards the other phase

Summary

Introduction

The development and designing of an efficient bioprocess is the most crucial step in any biotechnology industry. The prime objective of studying ATPS here is to replace and integrate various unit operations involved in fermentative production of pullulan, with the single stage aqueous partitioning. Extractive fermentation or in situ product removal, is a novel process deriving from ATPS where the microbial system grows in one phase and the product of interest being extracellular in nature is excreted out and partitioned towards the other phase. The use of ATPS in pullulan recovery and partial purification has been previously described, but extractive fermentation has not yet been employed for the exopolysaccharide production[20]. The effect of dextran, phosphate salt concentration and various PEG molecular mass upon microbial growth and pullulan yield was considered to select the appropriate ATPS system for in situ recovery of pullulan

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