Chemical glycosidation of trypsin with O-carboxymethyl-poly-beta-cyclodextrin: catalytic and stability properties

Biotechnol Appl Biochem. 2005 Jun;41(Pt 3):217-23. doi: 10.1042/BA20040115.

Abstract

The polysaccharide O-carboxymethyl-poly-beta-cyclodextrin was synthesized (molecular mass 13,000 Da, 40% carboxy groups) and attached to the surface of bovine pancreatic trypsin. The resulting neoglycoenzyme retained high proteolytic and esterolytic activity and contained approx. 1.0 mol of polymer/mol of enzyme. The optimum temperature for trypsin activity was increased by 10 degrees C after this transformation. Thermostability of the polymer-enzyme complex was increased by about 14 degrees C over 10 min incubation. The conjugate was also more resistant to thermal inactivation at different temperatures, ranging from 45 to 60 degrees C, demonstrating the influence of supramolecular and polymer-protein electrostatic interactions on trypsin thermostabilization. Additionally, the conjugate was 36-fold more resistant to the action of the anionic surfactant SDS. This modification also protected the enzyme from autolysis at alkaline pH.

Publication types

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

MeSH terms

  • Animals
  • Carboxymethylcellulose Sodium / chemistry*
  • Catalysis
  • Cattle
  • Enzyme Activation
  • Enzyme Stability
  • Hydrogen-Ion Concentration
  • Kinetics
  • Macromolecular Substances
  • Molecular Weight
  • Pancreas / enzymology
  • Protein Binding
  • Protein Denaturation
  • Static Electricity
  • Temperature
  • Trypsin / chemical synthesis
  • Trypsin / chemistry*
  • beta-Cyclodextrins / chemistry*

Substances

  • Macromolecular Substances
  • beta-Cyclodextrins
  • Trypsin
  • Carboxymethylcellulose Sodium