Golestan University of Medical Sciences Repository

Investigation of chromatography and polymer/salt aqueous two-phase processes for downstream processing development of recombinant phenylalanine dehydrogenase

Omidinia, E. and Shahbaz Mohamadi, H. and Dinarvand, R. and Taherkhani, H.-A. (2010) Investigation of chromatography and polymer/salt aqueous two-phase processes for downstream processing development of recombinant phenylalanine dehydrogenase. Bioprocess and Biosystems Engineering, 33 (3). pp. 317-329. ISSN 16157591 (ISSN)

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Abstract

This work presents a comprehensive study between the polymer/salt aqueous two-phase systems (ATPS) and chromatography process for downstream processing of recombinant Bacillus badius phenylalanine dehydrogenase (PheDH). First, the partitioning behavior of recombinant PheDH in polyethylene glycol (PEG)/K 2HPO4 ATPS was examined. For comparative purpose, a classical chromatographic protocol was performed as well. Investigation of chromatography and ATPS procedures revealed that the ATPS comprising of 9% (w/w) PEG-6000, 16% (w/w) K2HPO4 and 16% (w/w) KCl with pH of 8.0, volume ratio (V R ) of 0.25, temperature of 25 °C and 40% (w/w) cell lysate ensured the most favorable approach for PheDH downstream process. A specific activity of 4,231.4 U/mg, a yield of 96.7% and a recovery of 162.0% were obtained. Furthermore, the shorter process time (4 vs. 48 h) and the lower total cost (4 vs. 20 €) were additionally features that confirmed the suitability of proposed technique. © 2009 Springer-Verlag.

Item Type: Article
Additional Information: Unmapped bibliographic data: LA - English [Field not mapped to EPrints] J2 - Bioprocess Biosyst. Eng. [Field not mapped to EPrints] C2 - 19495799 [Field not mapped to EPrints] AD - Department of Biochemistry, Pasteur Institute of Iran, Tehran 13164, Iran [Field not mapped to EPrints] AD - Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran [Field not mapped to EPrints] AD - Faculty of Medicine, Golestan University of Medical Sciences, Gorgan, Iran [Field not mapped to EPrints] DB - Scopus [Field not mapped to EPrints]
Uncontrolled Keywords: Aqueous two-phase systems (ATPS), Chromatography, Downstream processing, Phenylalanine dehydrogenase (PheDH), Aqueous two phase, Aqueous two phase system, Aqueous two-phase systems (ATPS), Cell lysates, Comprehensive studies, Downstream process, Downstream-processing, Phenylalanine dehydrogenase, Process time, Recombinant bacilli, Specific activity, Total costs, Volume ratio, Amino acids, Bacteriology, Chromatographic analysis, Chromatography, Polyethylene oxides, Polyethylene glycols, bacterial enzyme, macrogol 6000, phenylalanine dehydrogenase, recombinant enzyme, unclassified drug, article, Bacillus, bacillus badius, biotechnological production, cell lysate, chromatography, comparative study, enzyme activity, enzyme purification, priority journal, Amino Acid Oxidoreductases, Biotechnology, Chromatography, Dose-Response Relationship, Drug, Escherichia coli, Hydrogen-Ion Concentration, Kinetics, Molecular Weight, Polyethylene Glycols, Polymers, Recombinant Proteins, Salts, Water, Bacillus badius
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
Divisions: معاونت تحقیقات و فناوری
Depositing User: GOUMS
Date Deposited: 15 Apr 2015 10:03
Last Modified: 17 Jan 2017 10:21
URI: http://eprints.goums.ac.ir/id/eprint/2218

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