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Use of p-aminophenyl D and L-lactic acids and p-aminophenyl pyruvic acid as effectors in the affinity chromatography of lactate dehydrogenase.
Authors:E Brown  R Joyeau
Institution:Laboratoire de Synthèse Organique, ERA 07.394, Faculté des Sciences, B.P. 535, 72017 Le Mans Cedex, France.
Abstract:p-Aminophenyl pyruvic acid and D-p-amino-phenyllactic acid were immobilized on a new synthetic acrylic carrier bearing acylating N-succinimidyl ester groups. The derivatives obtained were used successfully to purify lactate dehydrogenase (LDH) by affinity chromatography, the elution being carried out by means of NADH or preferably L-phenyllactic acid. Moreover, the specific activity of the LDH contained in a human blood serum was increased 270 times, using L-p-aminophenyllactic acid immobilized on a mixed polyacrylic agarose carrier.
Keywords:LDH  MDH  malate dehydrogenase  (or NADH)  oxidized (or reduced) nicotinamide adenine dinucleotide  TNBS  2  4  6-trinitrobenzene sulphonic acid  BSA  bovine serumalbumin  APL  APP  ACAPROSUC  cross-linked copolymer of acrylamide and (6-acrylamidohexanoyloxy) succinimide  BIS  AB-A6-NH  copolymer of agarose  BIS and 6-acrylamido hexanoic acid  AB-MGG  copolymer of agarose  BIS and methacryloyl glycylglycine  U (or mU)  activity unit (or milliunit) of dehydrogenase  one unit being the quantity of dehydrogenase which catalyzes the oxidation of 1 μmol NADH per minute at 25°C
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