Compositions and methods for enzyme immobilization
AssigneeAgroSpheres, Inc.
InventorsAmeer Hamza Shakeel, Zachery George Davis, Joseph Thomas Frank, Sepehr Zomorodi, Payam Pourtaheri
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Abstract
The present disclosure relates to compositions of immobilized enzymes on the surface of achromosomal and/or anucleate cells and uses thereof. In particular, the present disclosure provides genetically engineered minicells with enzymes self-assembled on their surface. The immobilized enzymes on the surface of achromosomal and/or anucleate minicells, has agricultural, industrial, and environmental applications due to their improved stability durability and, reusability. Also, provided are methods for producing and purifying enzyme-immobilized minicells.
Claims21
A method for improving activity and stability of enzyme-substrate binding, comprising:
- applying to a substrate an anucleated cell-based enzyme immobilization and delivery platform,
- wherein the anucleated cell-based enzyme immobilization and delivery platform comprises an anucleated cell that is derived from a protease deficient parental cell, and wherein the anucleated cell comprises (i) an expressed self-assembled enzyme immobilized to the surface of said cell and (ii) at least one fusion protein comprising at least one surface expressing moiety and at least one cell adhesion moiety, wherein the expressed self-assembled enzyme is fused with a linker domain, and
- wherein the activity and stability of enzyme-substrate binding is improved with the application of the anucleated cell when comparing to the application of an anucleated cell not comprising (i) the expressed self-assembled enzyme and (ii) the at least one fusion protein.
A method for improving activity and stability of enzyme-substrate binding, comprising:
- applying to a substrate an anucleated cell-based enzyme immobilization and delivery platform, wherein the anucleated cell-based enzyme immobilization and delivery platform comprises an anucleated cell that is derived from a protease deficient parental cell, wherein the anucleated cell comprises at least two different expressed self-assembled enzymes immobilized to a surface of said cell, wherein each of the expressed self-assembled enzymes is a fusion protein comprising at least one surface expressing moiety and at least one enzymatically active moiety, wherein said enzymatically active moiety of a first expressed self-assembled enzyme is lipase and said enzymatically active moiety of a second expressed self-assembled enzyme is not lipase, wherein each of the at least two different expressed self-assembled enzymes is fused with a linker domain, and wherein the activity and stability of enzyme-substrate binding is improved with the application of the anucleated cell when comparing to the application of an anucleated cell not comprising the at least two different expressed self-assembled enzymes.
A method for improving activity and stability of enzyme-substrate binding, comprising:
- applying to a substrate an anucleated cell-based enzyme immobilization and delivery platform, wherein the anucleated cell-based enzyme immobilization and delivery platform comprises an anucleated cell that is derived from a protease deficient parental cell, wherein the anucleated cell comprises at least two different expressed self-assembled enzymes immobilized to a surface of said cell, wherein each of the expressed self-assembled enzymes is a fusion protein comprising at least one surface expressing moiety and at least one enzymatically active moiety, wherein said enzymatically active moiety of a first expressed self-assembled enzyme is glucose isomerase and said enzymatically active moiety of a second expressed self-assembled enzyme is not glucose isomerase, wherein each of the at least two different expressed self-assembled enzymes is fused with a linker domain, and wherein the activity and stability of enzyme-substrate binding is improved with the application of the anucleated cell when comparing to the application of an anucleated cell not comprising the at least two different expressed self-assembled enzymes.
A method for improving activity and stability of enzyme-substrate binding, comprising:
- applying to a substrate an anucleated cell-based enzyme immobilization and delivery platform, wherein the anucleated cell-based enzyme immobilization and delivery platform comprises an anucleated cell that is derived from a protease deficient parental cell, wherein the anucleated cell comprises at least two different expressed self-assembled enzymes immobilized to a surface of said cell, wherein each of the expressed self-assembled enzymes is a fusion protein comprising at least one surface expressing moiety and at least one enzymatically active moiety, wherein said enzymatically active moiety of a first expressed self-assembled enzyme is protease and said enzymatically active moiety of a second expressed self-assembled enzyme is not protease, wherein each of the at least two different expressed self-assembled enzymes is fused with a linker domain, and wherein the activity and stability of enzyme-substrate binding is improved with the application of the anucleated cell when comparing to the application of an anucleated cell not comprising the at least two different expressed self-assembled enzymes.
Dependencies are inferred from the claim text.