Literature DB >> 1747122

Purification and characterization of cephalosporin 7 alpha-hydroxylase from Streptomyces clavuligerus.

X F Xiao1, S Wolfe, A L Demain.   

Abstract

Cephalosporin 7 alpha-hydroxylase, which catalyses the conversion of cephalosporins into their 7 alpha-hydroxy derivatives, was purified nearly 390-fold from Streptomyces clavuligerus through ion-exchange chromatography, (NH4)2SO4 fractionation, gel filtration and dye chromatography, with the use of h.p.l.c. to monitor enzyme activity. The nearly pure enzyme migrates as a single major band, with an Mr of 32,000 in SDS/PAGE. Its optimum pH is in the range 7.3-7.7. Under our conditions the reaction was fastest at temperatures in the range 20-30 degrees C. The Km for cephalosporin C is 0.72 mM, and the Vmax. is 15.4 mumol of cephalosporin C hydroxylated/min per mg. Cephalosporin 7 alpha-hydroxylase did not show any deacetoxycephalosporin C synthase or deacetoxycephalosporin C hydroxylase activity.

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Year:  1991        PMID： 1747122      PMCID： PMC1130572          DOI： 10.1042/bj2800471

Source DB:  PubMed          Journal:  Biochem J        ISSN： 0264-6021            Impact factor:   3.857


  8 in total

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Authors:  M M Bradford
Journal:  Anal Biochem       Date:  1976-05-07       Impact factor: 3.365

2.  Cleavage of structural proteins during the assembly of the head of bacteriophage T4.

Authors:  U K Laemmli
Journal:  Nature       Date:  1970-08-15       Impact factor: 49.962

3.  Deacetoxycephalosporin C synthetase and deacetoxycephalosporin C hydroxylase are two separate enzymes in Streptomyces clavuligerus.

Authors:  S E Jensen; D W Westlake; S Wolfe
Journal:  J Antibiot (Tokyo)       Date:  1985-02       Impact factor: 2.649

4.  Partial purification and catalytic properties of a bifunctional enzyme in the biosynthetic pathway of beta-lactams in Cephalosporium acremonium.

Authors:  A Scheidegger; M T Küenzi; J Nüesch
Journal:  J Antibiot (Tokyo)       Date:  1984-05       Impact factor: 2.649

5.  The conversion of cephalosporins to 7 alpha-methoxycephalosporins by cell-free extracts of Streptomyces clavuligerus.

Authors:  J O'Sullivan; E P Abraham
Journal:  Biochem J       Date:  1980-02-15       Impact factor: 3.857

6.  Purification and characterization of a 2-oxoglutarate-linked ATP-independent deacetoxycephalosporin C synthase of Streptomyces lactamdurans.

Authors:  J Cortés; J F Martín; J M Castro; L Láiz; P Liras
Journal:  J Gen Microbiol       Date:  1987-11

7.  Characterization and complementation of a cephalosporin-deficient mutant of Streptomyces clavuligerus NRRL 3585.

Authors:  J Piret; B Resendiz; B Mahro; J Y Zhang; E Serpe; J Romero; N Connors; A L Demain
Journal:  Appl Microbiol Biotechnol       Date:  1990-02       Impact factor: 4.813

8.  Metal ion-promoted binding of proteins to immobilized triazine dye affinity adsorbents.

Authors:  P Hughes; C R Lowe; R F Sherwood
Journal:  Biochim Biophys Acta       Date:  1982-01-04
  8 in total
  5 in total

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2.  A two-protein component 7 alpha-cephem-methoxylase encoded by two genes of the cephamycin C cluster converts cephalosporin C to 7-methoxycephalosporin C.

Authors:  J J Coque; F J Enguita; J F Martín; P Liras
Journal:  J Bacteriol       Date:  1995-04       Impact factor: 3.490

3.  Characterization of the cefF gene of Nocardia lactamdurans encoding a 3'-methylcephem hydroxylase different from the 7-cephem hydroxylase.

Authors:  J J Coque; F J Enguita; R E Cardoza; J F Martín; P Liras
Journal:  Appl Microbiol Biotechnol       Date:  1996-01       Impact factor: 4.813

4.  Cloning of a Streptomyces clavuligerus DNA fragment encoding the cephalosporin 7 alpha-hydroxylase and its expression in Streptomyces lividans.

Authors:  X Xiao; G Hintermann; A Häusler; P J Barker; F Foor; A L Demain; J Piret
Journal:  Antimicrob Agents Chemother       Date:  1993-01       Impact factor: 5.191

Review 5.  Genes for beta-lactam antibiotic biosynthesis.

Authors:  J F Martín; S Gutiérrez
Journal:  Antonie Van Leeuwenhoek       Date:  1995       Impact factor: 2.271
  5 in total

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