Probing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin C synthase.
Deacetoxycephalosporin C synthase from Streptomyces clavuligerus catalyses the conversion of the five-membered penicillin ring to the unsaturated six-membered cephem ring of deacetoxycephalosporin C. The effects on enzyme activity of the penicillin substrate sidechain and various cofactors were inve...
| Main Authors: | , , , , , |
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| Format: | Journal article |
| Sprog: | English |
| Udgivet: |
2001
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| _version_ | 1826286891189665792 |
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| author | Dubus, A Lloyd, MD Lee, H Schofield, C Baldwin, J Frere, J |
| author_facet | Dubus, A Lloyd, MD Lee, H Schofield, C Baldwin, J Frere, J |
| author_sort | Dubus, A |
| collection | OXFORD |
| description | Deacetoxycephalosporin C synthase from Streptomyces clavuligerus catalyses the conversion of the five-membered penicillin ring to the unsaturated six-membered cephem ring of deacetoxycephalosporin C. The effects on enzyme activity of the penicillin substrate sidechain and various cofactors were investigated using a continuous spectrophotometric assay. The conversion of penicillin G to phenylacetyl-7-aminodeacetoxycephalo sporanic acid (G-7-ADCA) was confirmed, and further details of the reaction were elucidated. The conversion of ampicillin to cephalexin was faster than that of acetyl-6-APA to acetyl-7-ADCA kcat = 0.120 +/- 0.001 s(-1) versus 0.035 +/- 0.001 s(-1), but they had similar Km values: 4.86 +/- 0.12 and 3.28 +/- 0.26 mM, respectively. Amoxycillin and penicillin V were also converted at low levels. Conversion was not detected for penicillanate, 6-aminopenicillanate, carbenicillin, temocillin, ticarcillin or benzylpenicilloic acid, suggesting that the enzyme has a relatively strict selectivity for the sidechain of the penicillin substrate. |
| first_indexed | 2024-03-07T01:50:27Z |
| format | Journal article |
| id | oxford-uuid:99e72f1e-d8a1-444f-8003-1c9e29ed0cf6 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T01:50:27Z |
| publishDate | 2001 |
| record_format | dspace |
| spelling | oxford-uuid:99e72f1e-d8a1-444f-8003-1c9e29ed0cf62022-03-27T00:17:41ZProbing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin C synthase.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:99e72f1e-d8a1-444f-8003-1c9e29ed0cf6EnglishSymplectic Elements at Oxford2001Dubus, ALloyd, MDLee, HSchofield, CBaldwin, JFrere, JDeacetoxycephalosporin C synthase from Streptomyces clavuligerus catalyses the conversion of the five-membered penicillin ring to the unsaturated six-membered cephem ring of deacetoxycephalosporin C. The effects on enzyme activity of the penicillin substrate sidechain and various cofactors were investigated using a continuous spectrophotometric assay. The conversion of penicillin G to phenylacetyl-7-aminodeacetoxycephalo sporanic acid (G-7-ADCA) was confirmed, and further details of the reaction were elucidated. The conversion of ampicillin to cephalexin was faster than that of acetyl-6-APA to acetyl-7-ADCA kcat = 0.120 +/- 0.001 s(-1) versus 0.035 +/- 0.001 s(-1), but they had similar Km values: 4.86 +/- 0.12 and 3.28 +/- 0.26 mM, respectively. Amoxycillin and penicillin V were also converted at low levels. Conversion was not detected for penicillanate, 6-aminopenicillanate, carbenicillin, temocillin, ticarcillin or benzylpenicilloic acid, suggesting that the enzyme has a relatively strict selectivity for the sidechain of the penicillin substrate. |
| spellingShingle | Dubus, A Lloyd, MD Lee, H Schofield, C Baldwin, J Frere, J Probing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin C synthase. |
| title | Probing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin C synthase. |
| title_full | Probing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin C synthase. |
| title_fullStr | Probing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin C synthase. |
| title_full_unstemmed | Probing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin C synthase. |
| title_short | Probing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin C synthase. |
| title_sort | probing the penicillin sidechain selectivity of recombinant deacetoxycephalosporin c synthase |
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