Ascorbic acid intermediates and process enzymes
AssigneeGenentech, Inc.
InventorsDavid R. Light, William H. Rastetter
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Abstract
2-keto-L-gluonic acid (2-KLG) is prepared from 2,5-diketo-D-gluconic acid (2,5-DKG) using a microorganism containing the enzyme 2,5-DKG reductase produced by an expression plasmid encoding a gene for the enzyme. The product, 2-KLG is a precursor for the synthesis of ascorbic acid (vitamin C).
Claims14
A process for converting 2,5-DKG into 2-KLG which comprises contacting a mixture comprising 2,5-DKG with a culture containing transformed host cells wherein the transformation with an expression vector containing the DNA sequence encoding 2,5-DKG-reductase renders the cell capable of expressing the gene for 2,5-DKG-reductase.
A cell or cell culture transformed with an expression vehicle containing the DNA sequence encoding 2,5-DKG reductase such that transformation renders the cell capable of effecting the expression of said 2,5-DKG reductase.
A recombinant expression vector capable of effecting the expression of a DNA sequence encoding 2,5-DKG reductase in suitable host cells.
A recombinant expression vehicle wherein a DNA sequence encoding 2,5-DKG reductase is operably linked to a promoter capable of effecting the expression of said DNA sequence.
A process for producing 2,5-DKG reductase which comprises culturing recombinant host cells containing a recombinant vector capable of effecting the expression of 2,5-DKG reductase.
A process for constructing a replicable expression vehicle capable of effecting the expression of a DNA sequence encoding 2,5-DKG reductase, which process comprises linking said DNA sequence operably to a promoter compatible with a bacterial host.
Plasmid pmit 12.
Plasmid pmit 12.DELTA..
Pasmid pmit 12.DELTA./trp.
Plasmid ptrp 1-35.
The DNA sequence of FIG. 4.
The DNA sequence of FIG. 4 comprising the 296th through the 1854th nuceleotide.
Dependencies are inferred from the claim text.