Stabilization of alpha-chymotrypsin by modification with beta-cyclodextrin derivatives

Biotechnol Appl Biochem. 2002 Dec;36(3):235-9. doi: 10.1042/ba20020056.

Abstract

Bovine pancreatic alpha -chymotrypsin was chemically modified with two different beta -cyclodextrin derivatives, named mono-6-formyl-beta-cyclodextrin and mono-6-succinyl-6-deoxy-beta-cyclodextrin. The modified enzymes contained approx. 3-5 mol of oligosaccharide/mol of protein, and retained full proteolytic and esterolytic activity. The optimum temperature for alpha -chymotrypsin was increased by 8 degrees C and its thermostability was enhanced by about 4-6 degrees C after modification. The conjugated enzymes were also more resistant to thermal inactivation at temperatures ranging from 45 to 55 degrees C. Additionally, the modified enzymes were 7-fold more stable against incubation at pH 9.0. The possible influence of supramolecular interactions on the thermal stabilization of modified alpha -chymotrypsins was also studied.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Chymotrypsin / chemistry*
  • Cyclodextrins / chemistry*
  • Enzyme Activation
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Kinetics
  • Macromolecular Substances
  • Pancreas / enzymology
  • Protein Conformation
  • Sensitivity and Specificity
  • Temperature
  • beta-Cyclodextrins*

Substances

  • Cyclodextrins
  • Macromolecular Substances
  • beta-Cyclodextrins
  • Chymotrypsin
  • alpha-chymotrypsin
  • betadex