Engineered ligase variants
AssigneeCodexis, Inc.
InventorsMathew G. Miller, Jonathan Vroom, Nikki Dellas, Donald S. Baskerville, Sandy M. Gomes, David Elgart, Judy Victoria Antonio Viduya
Life of this patent
Estimating the term…
Abstract
The present invention provides engineered ligase polypeptides and compositions thereof, as well as polynucleotides encoding the engineered ligase polypeptides. The invention also provides methods for use of the compositions comprising the engineered ligase polypeptides for diagnostic and other purposes.
Claims15
1-74. (canceled)
An engineered ligase comprising a polypeptide sequence having at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more sequence identity to a reference sequence, wherein the reference sequence is one of SEQ ID NOs:2, 6, 32, 34, and/or 38,
- wherein the engineered ligase comprises a 451K or 451G substitution in its polypeptide sequence, wherein the amino acid positions of the polypeptide sequence are numbered with reference to SEQ ID NO:6, and
- wherein the engineered ligase exhibits increased enzymatic activity of at least 1.1-fold, at least 2-fold, or at least 5-fold the enzymatic activity of a wild type T4 DNA ligase.
An engineered ligase, wherein the engineered ligase comprises a polypeptide sequence, wherein:
- (a) the polypeptide sequence has at least 85% sequence identity to a reference sequence of SEQ ID NO:6;
- and
- (b) the polypeptide sequence comprises the following amino acid sequence substitution, relative to the reference sequence of SEQ ID NO:6:
- T451K.
An engineered ligase, wherein the engineered ligase comprises a polypeptide sequences, wherein:
- (a) the polypeptide sequence has at least 85% sequence identity to a reference sequence of SEQ ID NO:6;
- and
- (b) the polypeptide sequence comprises the following amino acid sequence substitutions, relative to the reference sequence of SEQ ID NO:6:
- T451K, E88R, E440K, Q19K, L63T, P127K, K225A, F233A, A237R, D371W, and D452P.
Dependencies are inferred from the claim text.