Glutaryl cephalosporin amidase from pseudomonas diminuta BS-203
AssigneeBristol-Myers Squibb Company
InventorsRoss Binder, Joanne L. Brown, Thomas J. Franceschini, William V. Burnett, Michael Politino, Suo Win Liu, Sean M. Tonzi
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Abstract
The invention provides a glutaryl 7-ACA amidase from Pseudomonas diminuta strain BS-203, which catalyzes the hydrolysis of 7-β-(4-carboxybutanamido)-cephalosporanic acid to 7-aminocephalosporanic acid and glutaric acid. The invention also provides nucleic acid sequences, vectors, and host cells useful in the production of this amidase. The glutaryl 7-ACA amidase can be used for the preparation of 7-aminocephalosporanic acid.
Claims42
An isolated nucleic acid that encodes the glutaryl 7-ACA amidase having the amino acid sequence of SEQ ID NO:
- 2.
An isolated nucleic acid of between 10 and 1722 nucleotides, which hybridizes under stringent conditions either to a DNA molecule consisting of SEQ ID NO:
- 1 or to a DNA molecule consisting of the complement of SEQ ID NO:
- 1.
A glutaryl 7-ACA amidase having the following characteristics:
- isolatable from Pseudomonas diminuta BS-203, catalyzes the hydrolysis of 7-β-(4-carboxybutanamido)-cephalosporanic acid to 7-aminocephalosporanic acid and glutaric acid, and is composed of two subunits having apparent molecular weights of 42 kd and 26 kd by SDS PAGE electrophoresis.
A method for preparing a glutaryl 7-ACA amidase from Pseudomonas diminuta BS-203, said method comprising
- (a) culturing the Pseudomonas diminuta BS-203 bacterium under aerobic conditions in a suitable medium;
- and
- (b) recovering from the obtained culture a protein fraction having glutaryl 7-ACA amidase activity.
Dependencies are inferred from the claim text.