Literature DB >> 3183613

Purification and characterization of the isopenicillin N synthase of Streptomyces lactamdurans.

J M Castro1, P Liras, L Laíz, J Cortés, J F Martín.   

Abstract

The isopenicillin N synthase (cyclase) of Streptomyces lactamdurans (syn. Nocardia lactamdurans) has been purified to near homogeneity as judged by SDS-PAGE and isoelectric focusing. This enzyme catalyses the oxidative cyclization of the tripeptide delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine to isopenicillin N. The enzyme required DTT, Fe2+ and oxygen and it was greatly stimulated by ascorbic acid. It was strongly inhibited by Co2+, Zn2+ and Mn2+. Optimal pH and temperature were 7.0 and 25 degrees C (with the assay conditions used), respectively. The apparent Km of isopenicillin N synthase for delta-(L-alpha-aminoadipyl)-L-cysteinyl-D-valine was 0.18 mM. The enzyme is a monomer with an Mr of 26,500 +/- 1000 and a pI of 6.55.

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Year:  1988        PMID： 3183613     DOI： 10.1099/00221287-134-1-133

Source DB:  PubMed          Journal:  J Gen Microbiol        ISSN： 0022-1287


  8 in total

1.  Efficient Transformation of the Cephamycin C Producer Nocardia lactamdurans and Development of Shuttle and Promoter-Probe Cloning Vectors.

Authors:  C V Kumar; J J Coque; J F Martín
Journal:  Appl Environ Microbiol       Date:  1994-11       Impact factor: 4.792

2.  Cloning and characterization of the isopenicillin N synthase gene of Streptomyces griseus NRRL 3851 and studies of expression and complementation of the cephamycin pathway in Streptomyces clavuligerus.

Authors:  M García-Domínguez; P Liras; J F Martín
Journal:  Antimicrob Agents Chemother       Date:  1991-01       Impact factor: 5.191

Review 3.  Expression of genes and processing of enzymes for the biosynthesis of penicillins and cephalosporins.

Authors:  J F Martín; S Gutiérrez; F J Fernández; J Velasco; F Fierro; A T Marcos; K Kosalkova
Journal:  Antonie Van Leeuwenhoek       Date:  1994       Impact factor: 2.271

4.  Glucokinase-deficient mutant of Penicillium chrysogenum is derepressed in glucose catabolite regulation of both beta-galactosidase and penicillin biosynthesis.

Authors:  J L Barredo; E Alvarez; J M Cantoral; B Diez; J F Martin
Journal:  Antimicrob Agents Chemother       Date:  1988-07       Impact factor: 5.191

5.  Cloning and comparative sequence analysis of the gene coding for isopenicillin N synthase in Streptomyces.

Authors:  D Shiffman; M Mevarech; S E Jensen; G Cohen; Y Aharonowitz
Journal:  Mol Gen Genet       Date:  1988-11

Review 6.  Clusters of genes for the biosynthesis of antibiotics: regulatory genes and overproduction of pharmaceuticals.

Authors:  J F Martin
Journal:  J Ind Microbiol       Date:  1992 Feb-Mar

Review 7.  Genes for beta-lactam antibiotic biosynthesis.

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

8.  Beta-lactam biosynthesis in a gram-negative eubacterium: purification and characterization of isopenicillin N synthase from Flavobacterium sp. strain SC 12.154.

Authors:  H Palissa; H von Döhren; H Kleinkauf; H H Ting; J E Baldwin
Journal:  J Bacteriol       Date:  1989-10       Impact factor: 3.490
  8 in total

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