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Isolation of polyhydroxyalkanoate from hydrolyzed Cells of Bacillus flexus using Aqueous Two-phase System Containing Polyethylene Glycol and Phosphate.

Divyashree, M. S. and Shamala, T. R. and Rastogi, N. K. (2009) Isolation of polyhydroxyalkanoate from hydrolyzed Cells of Bacillus flexus using Aqueous Two-phase System Containing Polyethylene Glycol and Phosphate. Biotechnology and Bioprocess Engineering, 14 (4). pp. 482-489.

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Abstract

Main objective of present work was to isolate polyhydroxyalkanoate (PHA) from cell lysate of Bacillus flexus by aqueous-aqueous two-phase system (ATPS). Selected ATPS having polyethylene glycol (12%, w/v) and potassium phosphate (9.7%, pH 8.0) containing cell lysate obtained by sonication or hypochlorite treatment of B. flexus biomass (1g%, dry weight), was held at 280 C for 30 min, which partitioned PHA into top PEG phase and residual cell materials into bottom phase. For enzymatic cell hydrolysis, Microbispora sp culture filtrate having protease (3U/ml) was mixed with B. flexus biomass and ATPS, incubated at 37o C for 2 h prior to phase separation. PHA recovered by centrifugation was 19-51% of cell dry weight, depending on the mode of cells disruption. Protease was recovered along with PHA in the PEG phase and showed 7-fold increase in activity. PHA was characterized by GC, FTIR and 1H NMR. Results indicated that ATPS can be used for the isolation of PHA from hydrolyzed bacterial cells and purified protease can be recovered as a by-product, in a single defined experiment. Results have indicated that ATPS can be successfully employed as a non-organic solvent method for the isolation of PHA

Item Type: Article
Uncontrolled Keywords: Polyhydroxyalkanoate isolation; Bacillus flexus; Microbispora sp; Protease; Aqueous two-phase system; PEG/Phosphate
Subjects: 600 Technology > 08 Food technology > 09 Food Microbiology
Divisions: Food Microbiology
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 17 Mar 2010 06:52
Last Modified: 17 Mar 2010 06:52
URI: http://ir.cftri.com/id/eprint/9397

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