Engineered 3-o-kinase variants and methods of use
AssigneeCodexis, Inc.
InventorsDavid Entwistle, Stephanie Marie Forgel, Anders Matthew Knight, Mikayla Krawczyk, Philip Provencher, Amani Shoubber, Jonathan Vroom, David Watts, Leland Wong
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Abstract
The present invention provides engineered 3′O-kinase polypeptides useful for construction of materials used in template-independent polynucleotide synthesis, as well as compositions and methods of utilizing these engineered polypeptides. The present disclosure also describes one-pot methods for conversion of a natural or modified nucleoside to a nucleoside tetraphosphate or NQP.
Claims64
An engineered 3′O-kinase 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 selected from SEQ ID NO:
- 10, 142, 372, 450, 496, 1042, 1180, 1412 1464, 1800, and 2078, or a functional fragment thereof, and one or more amino acid residue differences relative to the reference sequence.
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A method of producing an NTP with a phosphate group at the 3′ position of the sugar (NQP), the method comprising (i) providing a 3′O-kinase enzyme, and (ii) contacting the 3′O-kinase enzyme with an NTP under suitable reaction conditions, such that an NQP is produced.
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Dependencies are inferred from the claim text.