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TitlePartial purification of penicillin acylase from Escherichia Coli in poly(ethylene glycol)-sodium citrate aqueous two-phase systems
Author(s)Marcos, João Carlos
Fonseca, Luís Pina
Ramalho, Maria Teresa
Cabral, J. M. S.
KeywordsAqueous two-phase systems
Poly(ethyleneglycol) - sodium citrate systems
Penicillin acylase
poly(ethylene glycol)-sodium citrate
Issue date1999
JournalJournal of Chromatography B
Citation"Journal of Chromatography B: Biomedical Sciences and Applications". ISSN 1570-0232. 734:1 (1999) 15-22.
Abstract(s)Studies on the partition and purification of penicillin acylase from osmotic shock extract Escherichia coli were performed in poly (ethylene glycol)- citrate systems. Both partition behavior of the enzyme and total protein are similar to those described in other reports increasing ,with pH and tie-line length and decreasing with PEG molecular weight . However, some selectivity could be attained with PEG 1000 systems and long tie-line at pH 6.9. In these conditions 2.6 fold purification with 83% yield were achieved. Influence of pH on partition shows that is the composition of the system and not the net charge of the enzyme that determine the behaviour in these conditions. Addition of NaCl to PEG 3350 systems significantly increases the partition of the enzyme. Althought protein partition also increased, purification conditions were possible with 1.5 M NaCl where 5.7 fold purification and 85% yield was obtained. This was possible due to the higher hydrophobicity of the enzyme compared to that of most of contaminants proteins.
AccessOpen access
Appears in Collections:CDQuim - Artigos (Papers)

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