Document Detail


Investigation of chromatography and polymer/salt aqueous two-phase processes for downstream processing development of recombinant phenylalanine dehydrogenase.
MedLine Citation:
PMID:  19495799     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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)/K2HPO4 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 degrees 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 euro) were additionally features that confirmed the suitability of proposed technique.
Authors:
Eskandar Omidinia; Hamid Shahbaz Mohamadi; Rassoul Dinarvand; Heshmat-Allah Taherkhani
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-06-04
Journal Detail:
Title:  Bioprocess and biosystems engineering     Volume:  33     ISSN:  1615-7605     ISO Abbreviation:  Bioprocess Biosyst Eng     Publication Date:  2010 Mar 
Date Detail:
Created Date:  2010-02-10     Completed Date:  2010-06-14     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101088505     Medline TA:  Bioprocess Biosyst Eng     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  317-29     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, Pasteur Institute of Iran, 13164, Tehran, Iran.
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Oxidoreductases / chemistry*
Biotechnology / methods*
Chromatography / methods*
Dose-Response Relationship, Drug
Escherichia coli / metabolism
Hydrogen-Ion Concentration
Kinetics
Molecular Weight
Polyethylene Glycols / chemistry
Polymers / chemistry*
Recombinant Proteins / chemistry*
Salts / chemistry
Water / chemistry
Chemical
Reg. No./Substance:
0/Polyethylene Glycols; 0/Polymers; 0/Recombinant Proteins; 0/Salts; 7732-18-5/Water; EC 1.4.-/Amino Acid Oxidoreductases; EC 1.4.3.-/phenylalanine oxidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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