Coupling of fructose-1,6-P2 to aminated agarose by Schiff base reduction. Affinity chromatography of yeast aldolase

Summary Fructose-1,6-P₂ was immobilized by sodium borohydride reduction of the Schiff base formed with aminated agarose (AH-Sepharose 4B^®). The coupling occurs with high yield (25 moles immobilized fructose-1,6-P₂ per ml packed gel) at neutral pH and room temperature. Schiff base reduction thus provides a convenient and mild coupling procedure for sugar phosphates preserving their labile phospho ester bonds. As exemplified by a new isolation procedure for fructose-1,6-P₂ aldolase from yeast, sugar phosphates insolubilized in this manner may be used for affinity chromatography of the corresponding enzymes, provided that contaminating unspecific phosphatases are removed in a preceding fractionation step.This work was supported by the Swiss National Science Foundation, grant No. 3.620-0.75. A preliminary account has been presented at the 10th Int. Congress of Biochemistry, Hamburg, 1976, Abstracts. p. 194.Acknowledgment. The excellent technical assistance of Alice Gasser is gratefully acknowledged