Simultaneous separation and purification of pyruvate kinase and lactate dehydrogenase by Dye-Ligand chromatography

dc.contributor.author	Makriyannis, T	en
dc.contributor.author	Clonis, YD	en
dc.date.accessioned	2014-06-06T06:42:29Z
dc.date.available	2014-06-06T06:42:29Z
dc.date.issued	1993	en
dc.identifier.issn	13595113	en
dc.identifier.uri	http://62.217.125.90/xmlui/handle/123456789/656
dc.relation.uri	http://www.scopus.com/inward/record.url?eid=2-s2.0-0000100701&partnerID=40&md5=cd5c8ba8318102cbe637b5dae5ce2c68	en
dc.title	Simultaneous separation and purification of pyruvate kinase and lactate dehydrogenase by Dye-Ligand chromatography	en
heal.type	journalArticle	en
heal.publicationDate	1993	en
heal.abstract	This work describes an improved downstream processing protocol for the simultaneous separation and purification of two diagnostic enzymes, lactate dehydrogenase (LDH) and pyruvate kinase (PK), from rabbit muscles. The purification scheme was specifically designed so that both purified enzymes were comparable, in terms of specific activity and impurity content, to commercial high-purity analytical grade. The proposed protocol comprises two dye-columns (dye-sorbents), Cibacron blue 3GA and Procion yellow MX-4G both immobilised on CL-Sepharose 6B, which were integrated in the following six-step scheme: (i) muscle extract preparation, (ii) 0-45% ammonium sulphate fractionation, (iii) blue-column chromatography where the two enzymes were separated and LDH was purified, (iv) blue-column chromatography, (v) thermal treatment, and (vi) yellow-column chromatography where PK was purified. This protocol affords LDH of specific activity 470 units/mg (no detectable PK) at 65% overall yield, and PK of specific activity 251 units / mg (no detectable LDH and enolase) at 40% overall yield.© 1993 Elsevier Science Publishers Ltd, England.	en
heal.journalName	Process Biochemistry	en
dc.identifier.issue	3	en
dc.identifier.volume	28	en
dc.identifier.spage	179	en
dc.identifier.epage	185	en