2,5-DKG permeases
AssigneeGenencor International, Inc.
InventorsVeronique A. Dartois, James A. Hoch, Fernando Valle, Manoj Kumar
Life of this patent
Estimating the term…
Abstract
The invention provides isolated nucleic acid molecules encoding polypeptides having 2,5-DKG permease activity, and oligonucleotides therefrom. The isolated nucleic acid molecules can be expressed in appropriate bacterial cells to enhance the production of 2-KLG, which can subsequently be converted to ascorbic acid. Further provided are isolated polypeptides having 2,5-DKG permease activity, immunogenic peptides therefrom, and antibodies specific therefor. The invention also provides methods of identifying novel 2,5-DKG permeases.
Claims36
An isolated nucleic acid molecule comprising the nucleic acid sequence of SEQ ID NO. 7.
An isolated nucleic acid molecule encoding a polypeptide having an amino acid sequence of SEQ ID NO. 8 or an amino acid sequence having at least 60% sequence identity to SEQ ID NO:
- 8, wherein said polypeptide has 2, 5-diketo-D-gluconlc acid (2,5-DKG) permease activity.
An isolated nucleic acid molecule comprising a nucleotide sequence having at least 40% sequence identity to SEQ ID NO:
- 7, wherein said nucleotide sequence encodes a polypeptide having 2,5-diketo-D-gluconic acid (2,5-DKG) permease activity.
An isolated oligonucleotide comprising at least 20 contiguous nucleotides of the nucleotide sequence of SEQ ID NO:
- 7, wherein said oligonucleotide is used as a probe and hybridizes under stringent hybridization conditions to a nucleic acid that encodes a polypeptide having 2,5-diketo-D-gluconic acid (2,5-DKG) permease activity.
A method of enhancing the production of 2-keto-L-gulonic acid (2-KLG) in a bacterial host comprising
- a) transforming a bacterial host with a nucleic acid which encodes a first polypeptide having 2,5-diketo-D-gluconic acid (2,5- DKG) permease activity, said first polypeptide having at least 60% amino acid sequence identity to SEQ ID NO:
- 8, wherein the bacterial host is capable of expressing an enzyme that catalyzes the conversion of 2,5-DKG to 2-KLG,
- b) culturing the transformed bacterial host under suitable culture conditions, and
- c) obtaining 2-KLG.
Dependencies are inferred from the claim text.