A spectrophotometric assay for deacetoxycephalosporin C synthase
Open Access
- 20 April 1987
- journal article
- Published by Wiley in FEBS Letters
- Vol. 214 (2) , 357-361
- https://doi.org/10.1016/0014-5793(87)80087-x
Abstract
A continuous direct spectrophotometric assay for deacetoxycephalosporin C synthase was developed, based on the absorption at 260 nm characteristic of the dihydrothiazine moiety of cephalosporins. K m values of 0.18 mM for penicillin N and 0.16 mM for α‐ketoglutarate were determined. A coupled assay using succinate thiokinase, pyruvate kinase and lactate dehydrogenase showed that succinate was a product of both deacetoxycephalosporin C synthase and hydroxylase reactions. The expandase reaction exhibited a 1:1.06 stoichiometry for deacetoxycephalosporin C and succinate.Keywords
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