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Downstream processing of phytase from Bacillus MCCB 242 employing aqueous two phase extraction
6th World Congress on Biotechnology
Sareen Sarah John, Ramya R Nair and I S Bright Singh
Cochin University of Science and Technology, India
Posters-Accepted Abstracts: J Biotechnol Biomater
DOI:
Abstract
Partitioning in aqueous two-phase systems (ATPS) is a powerful versatile downstream process and has great potential for large scale continuous separation of proteins and removal of contaminants from fermentation broths, since they produce an initial purification at a fast pace. Over the years, phytase has received tremendous importance making it the most preferred feed additive mainly for the reason that it is eco-friendly. To satisfy the need for efficient, easier, scalable and economically feasible downstream process for phytase production, aqueous two phase system (liquid-liquid extraction method) was adopted in this study. The purification of Bacillus MCCB 242 phytase using salt precipitation and ion-exchange chromatography has been studied and compared with aqueous two-phase system. An efficient recovery approach of aqueous two-phase systems (ATPS) with polyethylene glycol (PEG) and sodium citrate system was developed for phytase extraction which resulted in one sided phytase partitioning with a purification factor of 5.88. When compared with multi-step and single-step procedure, a phytase recovery of 78.5% within a short span (2 hours) was attained by ATPS whereas only 62% recovery could be obtained in 96 hours by chromatography. The purified phytase from ATPS was analyzed by SDS PAGE and a single band corresponding to molecular mass of 43 kDa was observed. Accordingly, ATPS was found to be an effective alternative for simultaneous partitioning and purification of phytase, hitherto not reported for recovery and purification of Bacillus phytase. In this context, outcome of the study is promising in developing an efficient industrial bio-process for phytase production.Biography
Sareen Sarah John has completed her MSc in Microbiology from Mahatma Gandhi University, Kerala and MPhil in Food & Industrial Microbiology from Annamalai University. She has submitted her PhD thesis entitled “Bioprocess optimization and characterization of phytase from an environmental isolate Bacillus MCCB 242” to Cochin University of Science & Technology, Kerala during December 2014. With 9 years experience in Microbiology/Biotechnology Research and Teaching, she has successfully completed 2 DST projects and has 2 journal publications, 13 abstract publications and won several national awards in paper/ presentations. Apart from her specialized interest in enzyme production, she is also interested in Genomics, Proteomics & Computational Biology.
Email: sareensj@yahoo.co.in
