A spectrophotometric assay for deacetoxycephalosporin C synthase

FEBS Letters

Crabbe, M.James C.¹  Baldwin, Jack E.¹ 

[1] The Dyson Perrins Laboratory, University of Oxford, South Parks Road, Oxford OX1 3QY, England

关键词: Cephalosporin Expandase;    Hydroxylase;    Succinate;    Enzyme kinetics;   

DOI  :  10.1016/0014-5793(87)80087-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown