Active site mutations of recombinant deacetoxycephalosporin C synthase

Biochemical and Biophysical Research Communications

Volumn 292, Issue 1, 2002, Pages 66-70

  Lee, Hwei Jen ^a,b   Schofield, Christopher J ^a   Lloyd, Matthew D ^a,c  

^a UNIVERSITY OF OXFORD   (United Kingdom)

^b NATIONAL DEFENSE MEDICAL CENTER   (Taiwan)

^c University of Bath   (United Kingdom)

Author keywords

2 Oxoglutarate dependent oxygenase; Antibiotic biosynthesis; Cephalosporins, DAOCS; Enzyme mechanism; Iron(II); Site directed mutagenesis

Indexed keywords

2 OXOGLUTARIC ACID; ADICILLIN; DEACETOXYCEPHALOSPORIN C SYNTHASE; PENICILLIN G; RECOMBINANT ENZYME; SYNTHETASE; UNCLASSIFIED DRUG;

ANALYTIC METHOD; ARTICLE; BINDING AFFINITY; CATALYSIS; CHEMICAL MODIFICATION; ENZYME ACTIVE SITE; IN VITRO STUDY; MUTATION; OXIDATION; PRIORITY JOURNAL; SITE DIRECTED MUTAGENESIS;

EID: 0036294889     PISSN: 0006291X     EISSN: None     Source Type: Journal    
DOI: 10.1006/bbrc.2002.6620     Document Type: Article

References (24)

1. Deacetoxycephalosporin C synthase and deacetylcephalosporin C synthase are two separate enzymes in Streptomyces clavuligerus
2. Purification and characterization of a 2-oxoglutarate-linked ATP-independent deacetoxycephalosporin C synthase of Streptomyces lactamdurans
3. Purification and initial characterization of deacetoxycephalosporin C synthase from Streptomyces clavuligerus
4. Studies on the active site of deacetoxycephalosporin C synthase
5. The iron(II) and 2-oxoacid-dependent dioxygenases and their role in metabolism
6. A dilemma of dioxygenases (or where biochemistry and molecular biology fail to meet)
7. Cloning and sequencing of the β-lactam hydroxylase gene (cefF) from Streptomyces clavu-ligerus: Gene duplication may have led to separate hydroxylase and expandase activities in the Actinomycetes
8. Copurification and characterization of deacetoxycephalosporin C synthetase/hydroxylase from Cephalosporium acremonium
9. Purification and initial characterization of an enzyme with deacetoxycephalosporin C synthetase and hydroxylase activities
10. Partial purification and catalytic properties of a bifunctional enzyme in the biosynthetic pathway of beta-lactams in Cephalosporium acremonium
11. Structure of a cephalosporin synthase
12. Probing the cosubstrate selectivity of deacetoxycephalosporin C synthase: The role of arginine-258
13. The role of arginine residues in substrate binding and catalysis by deacetoxycephalosporin C synthase
14. Kinetic and crystallographic studies on deacetoxycephalosporin C synthase (DAOCS)
15. Site-directed mutagenesis of virtually any plasmid by eliminating a unique site
16. Rapid and efficient site-specific mutagenesis without phenotypic selection
17. Rapid and efficient site-specific mutagenesis without phenotypic selection
18. The effect of cysteine mutations on the activity of recombinant deacetoxycephalosporin C synthase from S. clavuligerus
19. Substrate selectivity studies on deacetoxycephalosporin C synthase using a direct spectrophotometric assay
20. Adipoyl-6-APA is a substrate for deacetoxycephalosporin C synthase (DAOCS)
21. Alternative reactivity of an alpha-ketoglutarate-dependent iron(II) oxygenase: Enzyme self hydroxylation
22. Mutation of N304 to leucine in Streptomyces clavuligerus deacetoxycephalosporin C synthase creates an enzyme with increased penicillin analogue conversion
23. Structure of isopenicillin N synthase complexed with substrate and the mechanism of penicillin formation