The influence of water on protease-catalyzed peptide synthesis in acetonitrile/water mixtures

Eur J Biochem. 1988 Nov 1;177(2):313-8. doi: 10.1111/j.1432-1033.1988.tb14378.x.


Protease-catalyzed peptide synthesis in acetonitrile/water mixtures, containing 0-90% water, was investigated. alpha-Chymotrypsin, as well as thermolysin, were deposited on solid supports, prior to exposure to the reaction media. Peptide syntheses were performed using both a kinetically controlled process (chymotrypsin) and an equilibrium-controlled synthesis (thermolysin). The activity of chymotrypsin decreased at low water contents. However, at low water contents (1-10%) hydrolytic side reactions were suppressed and high yields of dipeptides were obtained. Optimal water content for the thermolysin-catalyzed reaction was 4-8%. The dipeptides produced were fully soluble in the reaction mixtures. High operational stability for alpha-chymotrypsin was obtained during 216 h of reaction.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetonitriles*
  • Adsorption
  • Catalysis
  • Chymotrypsin
  • Dipeptides / chemical synthesis
  • Drug Stability
  • Hydrolysis
  • Indicators and Reagents
  • Kinetics
  • Peptide Hydrolases / metabolism*
  • Peptides / chemical synthesis*
  • Solubility
  • Solutions
  • Thermolysin / metabolism
  • Water*


  • Acetonitriles
  • Dipeptides
  • Indicators and Reagents
  • Peptides
  • Solutions
  • Water
  • Peptide Hydrolases
  • Chymotrypsin
  • Thermolysin
  • acetonitrile