Effect of glycerol on thermolysin-catalyzed peptide bond synthesis.
The effect of glycerol on the hydrolytic activity of thermolysin (EC 3.4.24.4) has been compared with the effect on the condensation of N-benzyloxycarbonyl-L-aspartic acid with L-phenylalanine methyl ester to form N-benzyloxycarbonyl-L-aspartyl-L-phenylalanine methyl ester (Z X Asp X Phe X OMe), the precursor to the sweet-tasting compound L-aspartyl-L-phenylalanine methyl ester. Hydrolytic activity was measured by the degradation of azocasein and furylacryloyl-L-glycyl-L-leucinamide. Increasing concentrations of glycerol reversibly inhibited the hydrolytic activity of the enzyme toward both substrates. The inclusion of glycerol in the synthetic medium facilitated the production of Z X Asp X Phe X OMe in a water-soluble system but reduced the initial rate of peptide synthesis. Glycerol stabilized thermolysin against thermal denaturation.
D002384:Catalysis; D004795:Enzyme Stability; D005990:Glycerol / Q000494:pharmacology*; D006868:Hydrolysis; D007700:Kinetics; D010452:Peptide Biosynthesis; D013696:Temperature; D013820:Thermolysin / Q000037:antagonists & inhibitors / Q000378:metabolism*
D013820:Thermolysin; D005990:Glycerol