Affinity isolation of a cold-adapted enzyme: lactate dehydrogenase from Bacillus psychrosaccharolyticus.
A simple, economical and rapid affinity chromatography procedure with dyes as the ligand has been described for the one-step purification of a cold-adapted lactate dehydrogenase. Non-specific elution of Procion blue H-ERD-modified Sepharose yielded homogeneous preparations of lactate dehydrogenase both in column based procedures and in batch wise operations. Low operational temperatures resulted in the enhanced binding of the enzyme to the blue dye. The dissociation constants of the enzyme-dye complexes were 7.2 +/- 0.2 microM and 11.2 +/- 0.2 microM at 5 degrees C and 20 degrees C respectively.
Nandakumar
R
R
Department of Biotechnology, Lund University, Sweden.
Mattiasson
B
B
D001407:Bacillus / Q000201:enzymology*; D002846:Chromatography, Affinity / Q000379:methods; D003080:Cold Temperature; D004396:Coloring Agents; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012685:Sepharose
D004396:Coloring Agents; D012685:Sepharose; D007770:L-Lactate Dehydrogenase