Inactivation of deacetoxycephalosporin C synthase in extracts of Streptomyces clavuligerus during bioconversion of penicillin G to deacetoxycephalosporin G

By using cell-free extracts of Streptomyces clavuligerus containing deacetoxycephalosporin C synthase, we can convert penicillin G into deacetoxycephalosporin G. The degree of bioconversion was low (below 1% based on charged substrate) and decreased as substrate concentration was increased. Formatio...

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Bibliographic Details
Published in:Enzyme and microbial technology 1999-09, Vol.25 (6), p.497-501
Main Authors: Adrio, José L, Cho, Hiroshi, Piret, Jacqueline M, Demain, Arnold L
Format: Article
Language:English
Subjects:
Antibiotics
Bacteria
Bioconversion
Bioconversions. Hemisynthesis
Biological and medical sciences
Biotechnology
Catalyst deactivation
Cephalosporins
Composition effects
Deacetoxycephalosporin G
Fundamental and applied biological sciences. Psychology
Methods. Procedures. Technologies
Penicillin G
Streptomyces clavuligerus
Substrates
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Summary:By using cell-free extracts of Streptomyces clavuligerus containing deacetoxycephalosporin C synthase, we can convert penicillin G into deacetoxycephalosporin G. The degree of bioconversion was low (below 1% based on charged substrate) and decreased as substrate concentration was increased. Formation of product occurred during the first 2 h and then ceased. Addition of increasing levels of cofactors and/or substrate at the 2-h point did not result in more product. Preincubation of the crude extracts in the presence of ferrous ions plus either ascorbate or α-ketoglutarate led to complete loss of activity. Such inactivation could not be reversed by catalase, superoxide dismutase, mannitol, thiourea, dimethylsulfoxide, dithiothreitol, or β-mercaptoethanol.
ISSN:0141-0229
1879-0909
DOI:10.1016/S0141-0229(99)00084-8