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US 8,597,901 B2Granted
2,5-diketo-L-gluconic acid reductases and methods of use
AssigneeDanisco US Inc.
InventorsMark Donnelly, William H. Eschenfeldt, Jonathan Trent
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Abstract
Described herein are novel nucleic acids, proteins and methods that can be used to provide new catalysts with desirable traits for industrial processes. In particular, novel reductases isolated from the environment using PCR methods are described.
Claims8
A method for identifying a 2,5-diketo-L-gluconic acid reductase comprising:
- a) isolating nucleic acid molecules having homology to 2,5-diketo-L-gluconic acid reductase gene sequences from environmental samples that contain nucleic acids;
- comprising:
- (i) designing a set of degenerate primers SEQ ID NO:
- 11 and SEQ ID NO:
- 12 based on known 2,5-diketo-L-gluconic acid reductase or morphine dehydrogenase-gene sequences from various organisms;
- (ii) amplifying the environmental sample nucleic acid using the degenerate primers in a suitable polymerase chain reaction;
- (iii) identifying and cloning the amplified 2,5-diketo-L-gluconic acid reductase sequences;
- and
- (iv) expressing the amplified, cloned 2,5-diketo-L-gluconic acid reductase sequences as protein molecules;
- and
- b) screening said molecules for 2,5-diketo-D-gluconic acid reductase activity, wherein said molecules having 2,5-diketo-D-gluconic acid reductase activity are identified as a 2,5-diketo-L-gluconic acid reductase.
8 more elements
Dependencies are inferred from the claim text.