Abstract
A derivative of pyridoxal 5'-phosphate (PALP) bund to Sepharose 4B through the OH-group at position 3 of PALP was prepared by reacting PALP with bromoacetamidohexyl-Sepharose at pH 6.5. A Sepharose-bound tryptophan-PALP complex was also derived from the Sepharose-bound PALP by reducing the Schiff base between Sepharose-bound PALP and tryptophan with NaBH_4. These Sepharose-bound derivatives of PALP were found to be very useful as bio-specific adsorbents for apo-tryptophanase : Apo-tryptophanase adsorbed on a column of Sepharose -bound PALP was eluted specifically with a mixture of potassium phosphate buffer (0.5M, pH 7.0) and PALP (1mM), thus being purified 8〜10 times; much more specific elution of apotryptophanase adsorbed on a column of Sepharose-bound tryptophan-PALP comp1ex was observed. The apoprotein was eluted only with a mixture of PALP, tryptophan and potassium phosphate buffer, being much more efficiently purified by the affinity chromatography on the column.
