Preparation and properties of thermoresponsive bioconjugates of trypsin.

Abstract

Covalent attachment of enzymes and other proteins to the smart polymer, poly(N-isopropylacrylamide) [poly (NIPAAm)], has been widely used as a method for the preparation of thermosensitive protein conjugates. In the present study, reversible soluble-insoluble polymer-enzyme conjugates were prepared by conjugating a copolymer of NIPAAm with 5-mol % of 6-acrylaminohexanoic acid to trypsin by the carbodiimide-NHS (N-hydroxysuccinimide) coupling method. Four bioconjugates with different units of enzyme coupled to the matrix were prepared. Increased enzymatic activity in terms of high effectiveness factor (in the range of 3-5) was found in the conjugates. Kinetic parameters for the immobilized and free enzyme were determined. The Vmax/Km value of the enzyme significantly increased on immobilization by the factors in the range of 12-28. The immobilized enzyme also showed stability to autolysis at 50 degrees C.

• Authors: Smita Raghava, Kalyani Mondal, Munishwar N Gupta, Pradeep Pareek, Dirk Kuckling
• Journal: Artificial cells, blood substitutes, and immobilization biotechnology (Artif Cells Blood Substit Immobil Biotechnol) Vol. 34 Issue 3 Pg. 323-36 ( 2006) ISSN: 1073-1199 [Print] England
• PMID: 16809133 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Acrylic Resins
• Biopolymers
• Enzymes, Immobilized
• Trypsin Inhibitors
• poly-N-isopropylacrylamide
• Trypsin

Topics

• Acrylic Resins (chemistry, isolation & purification)
• Animals
• Autolysis
• Biopolymers (chemistry, isolation & purification)
• Cattle
• Enzyme Stability
• Enzymes, Immobilized (chemistry, isolation & purification, metabolism)
• Hot Temperature
• Hydrogen-Ion Concentration
• In Vitro Techniques
• Kinetics
• Materials Testing
• Spectrometry, Fluorescence
• Trypsin (chemistry, isolation & purification, metabolism)
• Trypsin Inhibitors