Intein-modified enzymes, their production and industrial application
First Claim
1. An intein modified protein comprising a target protein and an intein inserted into the target protein at a site i) immediately prior to a cysteine, threonine, or serine residue of the target protein, ii) within ten angstroms from an active site of the target protein, and iii) within 10 amino acids of a loop-β
- -sheet junction or within 10 amino acids of a loop-α
-helix junction of the target protein, wherein the target protein is a xylanase or a cellulase, and the intein has a sequence with at least 90% identity to a sequence selected from the group consisting of SEQ ID NOS;
3, 88, 91-96, 98, 99 and 101.
2 Assignments
0 Petitions
Accused Products
Abstract
A method of predicting an intein insertion site in a protein that will lead to a switching phenotype is provided. The method includes identifying a plurality of C/T/S sites within the protein; selecting from the plurality of C/T/S/sites those that are ranked 0.75 or higher by a support vector machine, within ten angstroms of the active site of the protein, and at or near a loop-β-sheet junction or a loop-α-helix junction. A method of controlling protein activity and hosts including proteins with controlled activity are also provided. Also, intein modified proteins and plants containing intein modified proteins are provided.
27 Citations
17 Claims
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1. An intein modified protein comprising a target protein and an intein inserted into the target protein at a site i) immediately prior to a cysteine, threonine, or serine residue of the target protein, ii) within ten angstroms from an active site of the target protein, and iii) within 10 amino acids of a loop-β
- -sheet junction or within 10 amino acids of a loop-α
-helix junction of the target protein, wherein the target protein is a xylanase or a cellulase, and the intein has a sequence with at least 90% identity to a sequence selected from the group consisting of SEQ ID NOS;
3, 88, 91-96, 98, 99 and 101. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
- -sheet junction or within 10 amino acids of a loop-α
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9. An intein modified protein having at least 90% identity to an amino acid sequence selected from the group consisting of SEQ ID NOS:
- 1692-1695, 1698, 1719, 1732, 1739-1742, 1745-1750, 1753-1766, 1768, 1769, 1772-1773.
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10. A transgenic plant including an expression construct encoding an intein modified protein comprising a target protein and an intein inserted into the target protein at a site i) immediately prior to a cysteine, threonine, or serine residue of the target protein, ii) within ten angstroms from an active site of the target protein, and iii) within 10 amino acids of a loop-α
- -sheet junction or within 10 amino acids of a loop-β
-helix junction of the target protein, wherein the target protein is a xylanase or a cellulase, and the intein has a sequence with at least 90% identity to a sequence selected from the group consisting of SEQ ID NOS;
3, 88, 91-96, 98, 99 and 101. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17)
- -sheet junction or within 10 amino acids of a loop-β
Specification