Affinity Chromatography of Yeast Alcohol Dehydrogenase Using Immobilized Monochlorotriazine Colourless Compounds
✍ Scribed by Yang Li; Gao Kunyu; Cheng Lubai; Zou Hanfa; Zhang Yukui
- Publisher
- John Wiley and Sons
- Year
- 1997
- Tongue
- English
- Weight
- 132 KB
- Volume
- 11
- Category
- Article
- ISSN
- 0269-3879
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✦ Synopsis
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.