Patent ID: 8597901

Claim:
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.