Alteration of the co-substrate selectivity of deacetoxycephalosporin C synthase: The role of arginine-258

Alteration of the 2-Oxoacid Cosubstrate Selectivity in Deacetoxycephalosporin C Synthase: The Role of Arginine-258

Studies on the Active Site of Deacetoxycephalosporin C Synthase

IRON(II) AND ALPHA-KETOISOVALERATE COMPLEX OF MUTANT ENZYME

MDTTVPTFSLAELQQGLHQDEFRRCLRDKGLFYLTDCGLTDTELKSAKDIVIDFFEHGSEAEKRAVTSPVPTMRRGFTGL
ESESTAQITNTGSYSDYSMCYSMGTADNLFPSGDFERIWTQYFDRQYTASRAVAREVLRATGTEPDGGVEAFLDCEPLLR
FRYFPQVPEHRSAEEQPLRMAPHYDLSMVTLIQQTPCANGFVSLQAEVGGAFTDLPYRPDAVLVFCGAIATLVTGGQVKA
PRHHVAAPRRDQIAGSSQTSSVFFLRPNADFTFSVPLARECGFDVSLDGETATFQDWIGGNYVNIRRTSKA

RECOMBINANT ENZYME OF S. CLAVULIGERUS ENZYME. R258Q MUTANT ENZYME

HANGING DROP AT 20 DEGREES C. 100 MM HEPES-NAOH, PH 7.0, 3% (W/V) GLYCEROL, 5 MM 2-OXO-3-METHYLBUTANOATE, 1.5-1.7 M AMMONIUM SULPHATE. CRYSTALS SOAKED IN 5 MM IRON(II) SULPHATE IN MOTHER LIQUOR UNDER ANAEROBIC CONDITIONS BEFORE DATA COLLECTION.

DEACETOXYCEPHALOSPORIN C SYNTHASE FROM S. CLAVULIGERUS

DELTA-R307A DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH SUCCINATE AND CARBON DIOXIDE

DELTA-R306 DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH IRON AND 2-OXOGLUTARATE.

DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH FE(II)

DEACETOXYCEPHALOSPORIN C SYNTHASE COMPLEXED WITH FE(II) AND 2-OXOGLUTARATE

TEXT TO EXPLAIN UNUSUAL UNIT-CELL DATA: R3 SPACE GROUP INDEX IN HEXAGONAL SETTING

CHAIN A ENGINEERED MUTATION ARG258GLN
DAOCS CATALYZES THE REACTION FROM PENICILLIN N TO DEACETOXY-
CEPHALOSPORIN C.

RESIDUE A50 BUILT AS ILE (AS REPORTED IN
PATENT) NOT LEU (AS REPORTED IN PAPER)
SIDECHAIN OF GLN-258 DISORDERED.

OXIDOREDUCTASE, ALTERNATIVE 2-OXOACIDS, CEPHEM ANTIBIOTIC BIOSYNTHESIS, CHEMICAL COSUBSTRATE RESCUE, CO-SUBSTRATE SELECTIVITY, 2- OXOGLUTARATE-DEPENDENT OXYGENASE