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.
-
Citations
17 Claims
-
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
-
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.
-
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