Method for refolding misfolded enzymes with detergent and cyclodextrin
First Claim
1. A method for folding an enzyme from a first misfolded configuration where the enzyme is completely defective in enzymatic activity to a second, native and active configuration where the enzyme has enzymatic activity, and then at least partially purifying the enzyme, the method comprising:
- adding a detergent having a linear alkyl non-polar portion to the enzyme to form a detergent-enzyme complex, where immediately prior to the addition of the detergent, the enzyme is in said first configuration;
contacting the detergent-enzyme complex with a cyclodextrin in an aqueous solution whereby the enzyme assumes the second active configuration; and
then at least partially purifying the enzyme that is in the second active configuration by separating it from the detergent.
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Abstract
A method for refolding an enzyme from a misfolded configuration to a second native and active configuration is presented. The method comprises adding a linear alkyl detergent to a misfolded enzyme to form an enzyme-detergent complex. Then the enzyme-detergent complex is contacted with a cyclodextrin to allow the enzyme to assume a second active configuration. The misfolded enzyme may have been previously denatured by heat or by chemical means other than a detergent.
23 Citations
7 Claims
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1. A method for folding an enzyme from a first misfolded configuration where the enzyme is completely defective in enzymatic activity to a second, native and active configuration where the enzyme has enzymatic activity, and then at least partially purifying the enzyme, the method comprising:
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adding a detergent having a linear alkyl non-polar portion to the enzyme to form a detergent-enzyme complex, where immediately prior to the addition of the detergent, the enzyme is in said first configuration; contacting the detergent-enzyme complex with a cyclodextrin in an aqueous solution whereby the enzyme assumes the second active configuration; and then at least partially purifying the enzyme that is in the second active configuration by separating it from the detergent. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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Specification