Immobilization of thermolysin for synthesis of aspartame precursor

Optimum conditions for immobilization of thermolysin, a metalloendopeptidase catalyzing synthesis of aspartame precursors, were investigated with using Amberlie XAD-7 as carrier and glutaraldehyde as cross-linking agent. Adsorption of thermolysin onto the carrier was rapid at the initial stage and 96% of the enzyme was adsorbed after 24 hours at . There was a linear relationship between amount of thermolysin adsorbed and thermolysin loaded upto 300g per liter of carrier. The effective range of cross-linking time, concentration of glutaraldehyde and pH for immobilization of the enzyme were , respectively. Degree of cross-linking and residual enzyme activity were high when cross-linked for 7 hours with 6% glutaraldehyde or for 3 hours with 12.5% glutaraldehyde. The residual enzyme activity was over 30% under these conditions.