title: Purification of lactic acid dehydrogenase from bovine heart crude extract by counter-current chromatography.
abstract: To test the utility of counter-current chromatography in purifying proteins, lactic acid dehydrogenase (LDH) was extracted from a crude bovine heart filtrate using a cross-axis coil planet centrifuge. The purification was performed with several polymer phase systems composed of 16% (w/w) poly(ethylene glycol) (PEG) 1000-12.5% (w/w) potassium phosphate buffers and 4.4% (w/w) PEG 8000-7.0 (w/w) dextran T500 at pH values ranging from 6.5 to 11.0. The best purification was achieved using PEG 1000-potassium phosphate buffer system at pH 7.3 by eluting the upper phase at 1.0 ml/min. Fractions were analyzed by LDH enzymatic activity and sodium dodecyl sulfate slab gel electrophoresis (SDS-PAGE). The LDH was purified directly from bovine heart crude extract within 3 h.
authors.affiliation: Department of Analytical Chemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Shibusawa
authors.forename: Y
authors.initials: Y
authors.lastname: Eriguchi
authors.forename: Y
authors.initials: Y
authors.lastname: Ito
mesh_terms: D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007770:L-Lactate Dehydrogenase / Q000302:isolation & purification* / Q000378:metabolism; D009206:Myocardium / Q000201:enzymology*; D014020:Tissue Extracts / Q000737:chemistry*
chemical_list: D014020:Tissue Extracts; D007770:L-Lactate Dehydrogenase