Protein engineering strategies to optimize activity of surface attached proteins
First Claim
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1. An isolated or recombinant enzyme comprising a plurality of artificial or recombinant surface coupling domains, wherein the enzyme, when coupled to a surface through the surface coupling domains, is enzymatically active.
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Abstract
Isolated and/or recombinant enzymes that include surface binding domains, surfaces with active enzymes bound to them and methods of coupling enzymes to surfaces are provided. Enzymes can include large and/or multiple surface coupling domains for surface coupling.
44 Citations
72 Claims
- 1. An isolated or recombinant enzyme comprising a plurality of artificial or recombinant surface coupling domains, wherein the enzyme, when coupled to a surface through the surface coupling domains, is enzymatically active.
- 19. A surface comprising an active enzyme bound thereon, wherein the enzyme is coupled to the surface through a plurality of artificial or recombinant surface coupling domains, and wherein the active enzyme displays a kcat/Km that is at least 10% as high as a corresponding active enzyme in solution.
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34. A method of binding an enzyme to a surface, the method comprising:
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providing an isolated or recombinant enzyme comprising a plurality of artificial or recombinant surface coupling domains; providing a surface comprising a plurality of binding partners that specifically bind to the surface coupling domains; contacting the enzyme to the surface; and
,permitting the binding partners to bind to the surface coupling domains, thereby binding the enzyme to the surface. - View Dependent Claims (35, 36, 37, 38)
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39. An isolated or recombinant active site-containing protein comprising:
- an artificial or recombinant surface coupling domain that is at least 5 kDa in size, wherein the protein, when coupled to a surface through the surface coupling domain, retains at least 1% activity as compared to an activity of a corresponding active protein in solution.
- View Dependent Claims (40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53)
- 54. A surface comprising a protein bound thereon, wherein the protein is coupled to the surface through an artificial or recombinant surface coupling domain that is at least 5 kDa in size, wherein the protein displays an activity that is at least 10% as high as a corresponding active protein in solution.
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68. A method of binding a protein to a surface, the method comprising:
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providing an isolated or recombinant protein comprising an artificial or recombinant surface coupling domain that is at least 5 kDa in size; providing a surface comprising a binding partner that specifically binds to the surface coupling domain; contacting the protein to the surface; and
,permitting the binding partner to bind to the surface coupling domain, thereby binding the enzyme to the surface; wherein the protein, when bound to the surface is at least 10% as active as the protein in solution. - View Dependent Claims (69, 70, 71, 72)
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Specification