Immobilized metal ion affinity chromatography on Co2+-carboxymethylaspartate-agarose Superflow, as demonstrated by one-step purification of lactate dehydrogenase from chicken breast muscle
G. Chaga et al., Immobilized metal ion affinity chromatography on Co2+-carboxymethylaspartate-agarose Superflow, as demonstrated by one-step purification of lactate dehydrogenase from chicken breast muscle, BIOT APP B, 29, 1999, pp. 19-24
A rapid method for the purification of lactate dehydrogenase from whole chi
cken muscle extract in one chromatographic step is reported. The purificati
on procedure can be accomplished in less than I h. A new type of immobilize
d metal ion affinity chromatography adsorbent is used that can be utilized
at linear flow rates higher than 5 cm/min, The final preparation of the enz
yme was with purity higher than 95 % as ascertained by SDS-PAGE, Three immo
bilized metal ions (Ni2+, Zn2+ and Co2+) were compared for their binding pr
operties towards the purified enzyme. The binding site of the enzyme for im
mobilized intermediate metal ions was determined after cleavage with CNBr a
nd binding studies of the derivative peptides on immobilized Co2+. A peptid
e located on the hi-terminus of the enzyme, implicated in the binding, has
great potential as a purification tag in fusion proteins.