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.
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