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.

Abstract: Echinocandin B deacylase is purified to near homogeneity from Actinoplanes utahensis by a process comprising, in order, extracting the soluble enzyme, heating to 60.degree. C., hydrophobic interaction chromatography, (NH.sub.4).sub.2 SO.sub.4 fractionation, gel filtration, cation exchange chromatography, dye-ligand chromatography, gel filtration, and cation-exchange chromatography.

Abstract: The present invention provides processes for isolating in substantially purified form water-soluble penicillin binding protein 2a of Staphylococcus aureus.

Abstract: Deacetoxycephalosporin C synthase obtained from cell-free extracts of Streptomyces clavuligerus was purified to near homogeneity and characterized in inactive form. The amino acid sequence was determined and the synthase recombinantly reproduced in active form in E. coli. The recombinantly produced synthase is provided in about 97% pure form by a chromatographic process. The synthase lacks DAOC hydroxylase function; however, it possesses the ability to transform 3-exomethylenocephalosporin C to deacetylcephalosporin C.

Abstract: Deacetoxycephalosporin C synthetase is provided in purified form via chromatography of crude cell-free extracts over a weak anion exchange resin followed by gel filtration and hydroxylapatite chromatography, all carried out in the presence of glycerol, a C.sub.1 -C.sub.3 alkyl monohydric alcohol, e.g., ethanol, a sulfhydryl containing reducing agent, e.g., dithiothreitol, and ascorbate. The purified enzyme which possesses both expandase and hydroxylase activities can be further purified by chromatography over a strong anion exchange resin.