Isolation of β-N-acetyl-d-hexosaminidases from lupin seed

A rapid procedure for the isolation of β-N-acetyl-d -hexosaminidase from lupin seed meal is described. This involves affinity chromatography of a seed extract on concanavalin A, followed by chromatography on DEAE-Sepharose. The purified enzyme was obtained in three forms, hexosaminidases A, B and B₁, capable of hydrolysing both p-nitrophenyl β-2-acetamido-2-deoxy-d-glucopyranoside and p-nitrophenyl β-2-acetamido-2-deoxy-d-galactopyranoside. Enzyme A was relatively less active towards the galactosaminide substrate, than were the B forms of the enzyme.