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AssigneeHoffmann-La Roche Inc.
InventorsAkiko Fujiwara, Tatsuo Hoshino, Masako Shinjoh
Estimating the term…
A novel coenzyme independent L-sorbosone dehydrogenase originating from a microorganism belonging to the genus Gluconobacter oxydans which acts on L-sorbosone to produce 2-keto-L-gulonic acid. The enzyme has the following physico-chemical properties: a) optimum pH: about 7.0, b) optimum temperature: about 30.degree. C. to about 40.degree. C., c) molecular sturucture: consisting of one type of unit having a molecular weight of about 47,500.+-.5,000 as measured by sodium dodecyl sulfate polyacrylamide gel electrophoresis, d) thermostability: stable below 30.degree. C., and e) inhibition: by Cu.sup.2+ -ions.
An isolated and purified DNA segment consisting essentially of a region encoding a Gluconobacter, coenzyme independent, L-sorbosone dehydrogenase, said dehydrogenase capable of converting L-sorbosone to 2-keto-L-gulonic acid.
A recombinant Gluconobacter microorganism transformed with a recombinant DNA vector, said vector having a DNA segment consisting essentially of a region encoding a Gluconobacter, coenzyme independent, L-sorbosone dehydrogenase said dehydrogenase capable of converting L-sorbosone to 2-keto-L-gulonic acid.
A process for producing 2-keto-L-gulonic acid comprising cultivating a Gluconobacter microorganism in a medium comprising a sugar selected from the group consisting of L-sorbose, L-sorbosone and D-sorbitol, wherein said microorganism is transformed with a recombinant DNA vector, said vector having a DNA segment consisting essentially of a region encoding a Gluconobacter, coenzyme independent, L-sorbosone dehydrogenase, said dehydrogenase capable of converting L-sorbosone to 2-L-gulonic acid.
A method for producing a transconjugant using a microorganism belonging to the genus Gluconobacter as a recipient, comprising contacting said recipient with a donor having a plasmid derived from pVK100 or pVK102, wherein said plasmid contains both a Mob site and cos site, and transferring the plasmid from the donor to the recipient through the use of the Tra gene function.
Dependencies are inferred from the claim text.