Method for recyclably using an enzyme
The invention provides methods and systems by which enzymes can be modified to improve solubility, catalytic activity, recoverability, and recyclability. The enzyme may be modified with a thermosensitive copolymer to form an enzyme-polymer conjugate that exhibits upper critical solution temperature (UCST) and/or lower critical solution temperature (LCST)-type behavior in an organic solvent, ionic liquid, or other solvent. Methods and systems of the invention facilitate the use of enzymes as biocatalysts in solvents.
1. A method for recyclably using an enzyme as a catalyst in a solvent, comprising:
a) reacting the enzyme with a polymer to form an enzyme-polymer conjugate;
b) dissolving the enzyme-polymer conjugate in the solvent to form a solution;
c) performing a chemical reaction in the solution, wherein the enzyme-polymer conjugate catalyzes the chemical reaction; and
d) extracting the enzyme-polymer conjugate from the solution,
wherein a catalytic activity of the enzyme-polymer conjugate after step d) is at least about 75% of a catalytic activity of the enzyme-polymer conjugate before step c).
2. The method of claim 1 , wherein the enzyme is lipase.
3. The method of claim 1 , further comprising purifying the enzyme-polymer conjugate by a method selected from the group consisting of affinity chromatography, ammonium sulfate precipitation, dialysis, and size exclusion chromatography.
4. The method of claim 1 , wherein the polymer is a copolymer.
5. The method of claim 4 , wherein the polymer is a random copolymer.
6. The method of claim 4 , wherein the copolymer comprises at least one monomer that is highly soluble, freely soluble, or soluble in the solvent and at least one monomer that is insoluble, poorly soluble, very slightly soluble, or slightly soluble in the solvent.
7. The method of claim 1 , wherein the chemical reaction is selected from the group consisting of a transesterification reaction, an oxidation/reduction reaction, a group transfer reaction, a hydrolysis reaction, an isomerization reaction, and a dehydration reaction.
8. The method of claim 1 , wherein step d) comprises changing a temperature of the solution to precipitate the enzyme-polymer conjugate.
9. The method of claim 1 , wherein step d) comprises extracting the enzyme-polymer conjugate with an extractant.
10. The method of claim 9 , wherein the enzyme-polymer conjugate exhibits upper critical solution temperature (UCST) behavior in the solvent and lower critical solution temperature (LCST) behavior in the extractant.
11. The method of claim 9 , wherein the enzyme-polymer conjugate exhibits LCST behavior in the solvent and UCST behavior in the extractant.