Transferases and oxidoreductases, nucleic acids encoding them and methods for making and using them

Patent metadata

Field	Value
Publication number	US-9399763-B2
Application number	US-201414182705-A
Country	US
Kind code	B2
Filing date	Feb 18, 2014
Priority date	Jan 3, 2008
Publication date	Jul 26, 2016
Grant date	Jul 26, 2016

Abstract

This invention relates generally to enzymes, polynucleotides encoding the enzymes, the use of such polynucleotides and polypeptides and more specifically to enzymes having transferase activity, e.g., transaminase activity, e.g., d-amino-acid transferase activity, and/or oxidoreductase activity, e.g., dehydrogenase activity, e.g., d-amino-acid dehydrogenase activity, and/or catalyze the transfer of a chemical group, catalyze transamination, catalyze the reaction: D-alanine+2-oxoglutarate<=>pyruvate+D-glutamate, and/or catalyze an oxidation-reduction reaction, catalyze the removal of hydrogen atoms, and/or catalyze the reaction: D-amino acid+H 2 O+acceptor<=>a 2-oxo acid+NH 3 +reduced acceptor.

First claim

What is claimed is:

1. An isolated, synthetic, or a recombinant nucleic acid encoding a polypeptide having a d-amino-acid transferase activity, wherein the polynucleotide is selected from the group consisting of:
(a) a variant nucleic acid sequence having 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity over the full length of a cDNA, transcript (mRNA) or gene to the nucleic acid sequence of SEQ ID NO: 869; and
(b) a nucleic acid encoding the polypeptide having d-amino-acid transferase activity, wherein the polypeptide comprises the amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the polypeptide of SEQ ID NO:870.

2. An expression cassette, a vector, or a cloning vehicle comprising the nucleic acid sequence of claim 1 , wherein optionally the cloning vehicle comprises a viral vector, a plasmid, a phage, a phagemid, a cosmid, a fosmid, a bacteriophage or an artificial chromosome.

3. The cloning vehicle of claim 2 , wherein the viral vector comprises an adenovirus vector, a retroviral vector or an adeno-associated viral vector, or, the artificial chromosome comprises a bacterial artificial chromosome (BAC), a bacteriophage Pl-derived vector (PAC), a yeast artificial chromosome (YAC), or a mammalian artificial chromosome (MAC).

4. A transformed cell comprising the expression cassette, vector or cloning vehicle of claim 2 , wherein the transformed cell is a bacterial cell, a mammalian cell, a fungal cell, a yeast cell, an insect cell, or a plant cell.

5. The polynucleotide of claim 1 , encoding a variant polypeptide having at least one conservative amino acid substitution and retaining d-amino-acid transferase activity; wherein the at least one conservative amino acid substitution comprises substituting an amino acid with another amino acid of like characteristics; replacement of an aliphatic amino acid with another aliphatic amino acid; replacement of a Serine with a Threonine, or vice versa; replacement of an acidic residue with another acidic residue; replacement of a residue bearing an amide group with another residue bearing an amide group; exchange of a basic residue with another basic residue; or replacement of an aromatic residue with another aromatic residue.

6. The polynucleotide of claim 1 , encoding the polypeptide having d-amino-acid transferase activity but lacking a signal sequence, a prepro domain, a binding domain.

7. The polynucleotide of claim 6 , wherein the binding domain consists of: a NAD, a NAD(P), a calcium, a thiamine, a FAD, a zinc, a DNA and a lipoyl binding domain.

8. The polynucleotide of claim 1 , encoding the polypeptide having d-amino-acid transferase and further comprising a heterologous sequence.

9. The polynucleotide of claim 8 , wherein the heterologous sequence consists of: a heterologous signal sequence, a heterologous domain, a heterologous binding domain, a heterologous dockerin domain, a heterologous catalytic domain (CD), or a combination thereof.

10. A nucleic acid sequence fully complementary to the nucleic acid sequence of claim 1 .

Assignees

• Verenium Corp
• Basf Enzymes Llc

Inventors

• Weiner David P
• Luginbuhl Peter
• Bueno Analia
• Cuenca Joslin M
• Marasco Erin

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• C12Q1/6876

  Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes · CPC title

• C12N15/1137

  against enzymes (viral enzymes C12N15/1131; receptors C12N15/1138) · CPC title

• C12N9/0006

  acting on CH-OH groups as donors (1.1) · CPC title

• A61K38/00

  Medicinal preparations containing peptides (peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00) · CPC title