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Simultaneous purification and isoelectric point (pI) determination was carried out at analytical scale of the chromosomal cephalosporinase from the Proteus vulgaris 1028 strain. Comparison of the enzyme to the purification results with m-aminophenylboronic acid-agarose affinity chromatography with sodium dodecyl sulphate-polyacrylamide gel electrophoresis revealed that minute amounts of accompanying proteins having identical pI values but different molecular masses were found in the chromatofocused preparation. The molecular mass of the enzyme was 24,000 dalton. The pI was found to be 8.3.

Although still there are Klebsiella strains which do not harbour plasmids and produce constitutive chromosomal beta-lactamases, recently clinical isolates were found in ever increasing numbers carrying mainly TEM-, CARB- and OXA type R-factors. We selected four chromosomal cephalosporinase producing Klebsiella strains to study the pI values of the enzymes and their simultaneous separability from accompanying proteins by chromatofocusing techniques. We compared pI values of the pure and the crude preparations: K. pneumoniae K1 SC 10436: pIpure = 6.4, pIcrude = 6.42; K. aerogenes K1 1082 E: pIpure = 6.5, pIcrude = 6.5; K. oxytoca 1082 E: pIpure = 6.42, pIcrude = 6.4; K. oxytoca 20: pIpure = 7.62, pIcrude = 7.6. Excellent agreement of the pI values among each other, but occasional differences with those obtained by analytical isoelectrofocusing are attributed to methodological diversities and to the presence of satellite enzymes, known to exist in Klebsiella.