title: Affinity chromatography of yeast alcohol dehydrogenase using immobilized monochlorotriazine colourless compounds.
abstract: Four new symmetrical colourless biomimetics bearing two monochlorotriazine rings have been rationally designed and synthesized. These immobilized colourless ligands and one analogue of Cibacron Blue F3GA on Sepharose CL-4B were used to purify alcohol dehydrogenase from baker's yeast extract. Twenty-two-fold purification with 78% enzyme recovery was achieved in a single step with specific elution of NAD+ (5 mM) from the colourless absorbent comprised of two ortho-aminobenzene sulphonates as terminal rings, which is comparable to the result obtained from the analogue of Cibacron Blue F3GA. Differential spectra between ligand-enzyme complexes and free ligands were studied.
authors.affiliation: National Chromatographic R & A Center, Dalian Institute of Chemical and Physics, Chinese Academy of Sciences, P.R. China.
authors.forename: Y
authors.initials: Y
authors.lastname: Li
authors.forename: G
authors.initials: G
authors.lastname: Kunyu
authors.forename: C
authors.initials: C
authors.lastname: Lubai
authors.forename: Z
authors.initials: Z
authors.lastname: Hanfa
authors.forename: Z
authors.initials: Z
authors.lastname: Yukui
mesh_terms: D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D002847:Chromatography, Agarose / Q000379:methods; D004396:Coloring Agents / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D009243:NAD / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000201:enzymology*; D014227:Triazines / Q000737:chemistry*
chemical_list: D004396:Coloring Agents; D008024:Ligands; D014227:Triazines; D009243:NAD; C012842:Cibacron Blue F 3GA; D000426:Alcohol Dehydrogenase