DEVELOPMENT OF AMIDASES FOR USE IN BETA-LACTAM MODIFICATION

Award Information
Agency:
Department of Health and Human Services
Branch
n/a
Amount:
$50,000.00
Award Year:
1988
Program:
SBIR
Phase:
Phase I
Contract:
n/a
Award Id:
8032
Agency Tracking Number:
8032
Solicitation Year:
n/a
Solicitation Topic Code:
n/a
Solicitation Number:
n/a
Small Business Information
1885 33rd St, Boulder, CO, 80301
Hubzone Owned:
N
Minority Owned:
N
Woman Owned:
N
Duns:
n/a
Principal Investigator:
() -
Business Contact:
() -
Research Institute:
n/a
Abstract
SEVERAL CLINICALLY IMPORTANT BETA-LACTAM ANTIBIOTICS ARE PRODUCED THROUGH CONDENSATION OF D-(-)-ALPHA-AMINOPHENYLACETIC ACID WITH PENAM AND CEPHEM NUCLEI IN MULTI-STEP CHEMICAL PROCESSES. AN EFFICIENT ENZYMATIC PROCESS FOR THE SYNTHESIS OF THESE ANTIBIOTICS WOULD LEAD TO INCREASED PRODUCT YIELDS AND DECREASED COST OFPRODUCTION. PREVIOUSLY DESCRIBED ENZYMES CATALYZE THE SYNTHESIS OF THESE ANTIBIOTICS AT RATES THAT ARE INADEQUATE FOR COMMERCIAL PROCESSES. THE AIM OF PHASE I RESEARCH IS TO DEMONSTRATE THAT ENZYMES ABLE TO CATALYZE THE SYNTHESIS OF THESE ANTIBIOTICS AT HIGH RATES CAN BE DERIVED FROM PENICILLIN AMIDASE. TO ACHIEVE THIS, MUTANTS WILL BE SELECTED FOLLOWING MUTAGENESIS OF CELLS THAT CARRY THE CLONED GENE FOR PENICILLIN AMIDASE BASED ON AN IMPROVED ABILITY TO HYDROLYZE A MODEL SUBSTRATE WITH A D-(-))-ALPHA-AMINOPHENYLACETYL MOIETY. THE RATES AT WHICH AMIDASES PURIFIED FROM THESE MUTANTS CATALYZE THE HYDROLYSISOF THE MODEL SUBSTRATE AND THE CONDENSATION OF D-(-)-ALPHA-AMINOPHENYLACETIC ACID WITH 6-AMINOPENICILLANIC ACID TO PRODUCE AMPICILLIN WILL BE DETERMINED AND COMPARED TO THE RATES OBSERVED WITH THE WILD-TYPE AMIDASE. THE AIM OF PHASE II RESEARCH WILL BE THE REITERATIVE USE OF THE METHODS DEVELOPED IN PHASE I TO OBTAIN ENZYMES ABLE TO SYNTHESIZE AMPICILLIN AND CEPHALEXIN AT COMMERCIALLY USEFUL RATES.

* information listed above is at the time of submission.

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