Kinetics of enzymatic solid-to-solid peptide synthesis: Intersubstrate compound, substrate ratio, and mixing effects

Markus Erbeldinger, Xiongwei Ni, Peter J. Halling

Research output: Contribution to journalArticle

Abstract

A systematic study of thermolysin-catalyzed solid-to-solid peptide synthesis using Z-Gln and Leu-NH2 as model substrates was carried out. The aim was to extend the kinetic knowledge of this new reaction system involving highly concentrated substrate mixtures with little water (10% to 20% w/w). Preheating of the substrates, and ultrasonic treatment, as described in the literature, had no significant effect on our system. The formation of a third compound, the salt of the two substrates, was discovered during melting point experiments. This was associated with a very strong dependence of kinetics on the exact substrate ratio (e.g., two-fold higher initial rate with 60% Leu-NH2 and 40% Z-Gln than with the equimolar substrate ratio). A model was developed to show how the composition and pH of the liquid phase depends on the substrate ratio, and seemed to explain the experimental rates. In addition, the influences of different mixing and water distribution methods are described. Finally, we can now summarize the major effects of the reaction system as a starting point for further research and scale-up studies.

Original languageEnglish
Pages (from-to)316-321
Number of pages6
JournalBiotechnology and Bioengineering
Volume63
Issue number3
DOIs
Publication statusPublished - 5 May 1999

Keywords

  • Enzymatic
  • Peptide synthesis
  • Proteases
  • Solid-to-solid conversion

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