Deacetoxycephalosporin C hydroxylase

Details Deacetoxycephalosporin C hydroxylase Deacetoxycephalosporin C hydroxylase

1992-04-23

1998-02-03

Knode, Marian C.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Oxidoreductase

C12N 902

Patent

active

057143633

ABSTRACT:
Deacetoxycephalosporin C hydroxylase is obtained in purified form from crude cell-free extracts via chromatography over a weak anionic exchange resin, ammonium sulfate fractionation, gel filtration, hydroxylapatite chromatography, and FPLC. The enzyme is obtained in >90% purity by further gel filtration and a second FPLC. The 28-residue amino-terminal sequence of hydroxylase is provided as well as a 9-residue amino-terminal sequence of an internal sequence and a 3-residue carboxy-terminal sequence. In addition to the efficient conversion of DAOC to DAC, the hydroxylase provided herein converts 7.beta.-(.alpha.-aminoadipamido)-3-exomethylenecepham-4-carboxylic acid to DAC.

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Baldwin et al., "Purification and Initial Characterization of an Enzyme with Deacrtoxycephalosporin C Synthetase and Hydroxylan Activities", Biochem J. 245:831-841 1987.
Dotzlaf et al., "Copurification and Characterization of Deacrtoxycephalosporin C Synthetase of Hydroxylase from Cephalosporin acremonium", J. Bacteriology 169 (4):1611-1618, 1987.
Jansen et al., "Deacetoxycephalosporin C . . . Streptomyces clavuligerus", J. Anitbodies, vol. 38, No. 2, pp. 263-265, 1985.
Scheidegger et al., "Partial Purification . . . Cephalosporium acremonium", J. Antibiotics, vol. 37, No. 5, pp. 522-531, 1984.

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