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Trypsin modification by vinyl polymers with variable solubilities in response to external signals
1Department of Polymer Chemistry, Faculty of Engineering, Kyoto University, Japan.
Bioconjugate Chemistry
|September 1, 1993
Summary
Enzyme immobilization was achieved using stimuli-responsive polymers. Modified trypsin showed reversible solubility and activity changes with pH, enabling controlled enzymatic reactions.
Area of Science:
- Biochemistry
- Polymer Chemistry
- Enzyme Engineering
Background:
- Enzyme immobilization is crucial for biocatalysis and enzyme recovery.
- Developing stimuli-responsive materials for controlled enzyme activity is an active research area.
- Graft polymerization offers a versatile method for modifying biomolecules with polymers.
Purpose of the Study:
- To synthesize and characterize trypsin modified with redox- and pH-sensitive vinyl polymers.
- To evaluate the solubility, stability, and enzymatic activity of the modified trypsin under different external stimuli.
- To explore the potential of these modified enzymes in controlled biocatalytic applications.
Main Methods:
- Graft polymerization of vinyl monomers onto a trypsin derivative initiated by aliphatic azo groups.
- Synthesis of redox-sensitive poly[3-carbamoyl-1-(p-vinylbenzyl)pyridinium chloride] and pH-sensitive poly(methacrylic acid-co-methyl methacrylate).
- Assessment of enzyme solubility and activity in response to redox agents (Na2S2O4, H2O2) and pH changes.
Main Results:
- Trypsin modified with redox-sensitive polymer exhibited reversible insolubility/solubility triggered by Na2S2O4 and H2O2, but activity was not proportional to enzyme concentration.
- Trypsin modified with pH-sensitive polymer showed reversible solubility changes with pH (insoluble in acidic, soluble in neutral conditions).
- The enzymatic activity of the pH-sensitive modified trypsin remained proportional to the enzyme concentration in homogeneous solutions, demonstrating reversible activity control.
Conclusions:
- Stimuli-responsive polymer modification provides a method for controlling enzyme solubility and activity.
- pH-sensitive polymers offer promising reversible control over enzyme activity and solubility for biocatalysis.
- Redox-sensitive polymers require further optimization to ensure proportional activity with enzyme concentration in solution.