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

Palissa, H.; Döhren, Hans von; Kleinkauf, Horst; Ting, Hong-Hoi; Baldwin, Jack E.

doi:10.1128/jb.171.10.5720-5728.1989

Cited by 20 publications

(6 citation statements)

References 47 publications

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“…The regulation of these CH␤Ls, if any, would be of interest as described for the A. hydrophila CH␤L (1). It may be hypothesized that CH␤Ls in C. meningosepticum may counteract the effect of antibiotics produced by this Chryseobacterium species (33,37). Additionally, the presence of two CH␤L genes in C. meningosepticum may be used as a tool for a PCR-based identification of this species.…”

Section: Discussionmentioning

confidence: 99%

Molecular and Biochemical Heterogeneity of Class B Carbapenem-Hydrolyzing β-Lactamases in Chryseobacterium meningosepticum

Bellais

Aubert

Naas

et al. 2000

Antimicrob Agents Chemother

132

134

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Although the carbapenem-hydrolyzing ␤-lactamase (CH␤L) BlaB-1 is known to be in Chryseobacterium meningosepticum NCTC 10585, a second CH␤L gene, bla GOB-1 , was cloned from another C. meningosepticum clinical isolate (PINT). The G؉C content of bla GOB-1 (36%) indicated the likely chromosomal origin of this gene. Its expression in Escherichia coli DH10B yields a mature CH␤L with a pI of 8.7 and a relative molecular mass of 28.2 kDa. In E. coli, GOB-1 conferred resistance to narrow-spectrum cephalosporins and reduced susceptibility to ureidopenicillins, broad-spectrum cephalosporins, and carbapenems. GOB-1 had a broadspectrum hydrolysis profile including penicillins and cephalosporins (but not aztreonam). The catalytic efficiency for meropenem was higher than for imipenem. GOB-1 had low amino acid identity with the class B CH␤Ls, sharing 18% with the closest, L-1 from Stenotrophomonas maltophilia, and only 11% with BlaB-1. Most of the conserved amino acids that may be involved in the active site of CH␤Ls (His-101, Asp-103, His-162, and His-225) were identified in GOB-1. Sequence heterogeneity was found for GOB-1-like and BlaB-1-like ␤-lactamases, having 90 to 100% and 86 to 100% amino acid identity, respectively, among 10 unrelated C. meningosepticum isolates. Each isolate had a GOB-1-like and a BlaB-1-like gene. The same combination of GOB-1-like and BlaB-1-like ␤-lactamases was not found in two different isolates. C. meningosepticum is a bacterial species with two types of unrelated chromosome-borne class B CH␤Ls that can be expressed in E. coli and, thus, may represent a clinical threat if spread in gram-negative aerobes.

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Section: Discussionmentioning

confidence: 99%

Molecular and Biochemical Heterogeneity of Class B Carbapenem-Hydrolyzing β-Lactamases in Chryseobacterium meningosepticum

Bellais

Aubert

Naas

et al. 2000

Antimicrob Agents Chemother

132

134

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“…IPNS was purified to homogeneity from A. chrysogenum (27,142,251) and has subsequently been obtained from P. chrysogenum (267), A. nidulans (339), several actinomycetes such as S. clavuligerus (153), and the gram-negative bacterium Flavobacterium sp. (250). It was shown that two interconvertable forms of the enzyme exist, an oxidized state with a disulfide linkage and a reduced state (27).…”

Section: Structural Genes and Deduced Proteinsmentioning

confidence: 99%

Molecular Regulation of β-Lactam Biosynthesis in Filamentous Fungi

Brakhage¹

1998

Microbiol Mol Biol Rev

211

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SUMMARY The most commonly used β-lactam antibiotics for the therapy of infectious diseases are penicillin and cephalosporin. Penicillin is produced as an end product by some fungi, most notably by Aspergillus (Emericella) nidulans and Penicillium chrysogenum. Cephalosporins are synthesized by both bacteria and fungi, e.g., by the fungus Acremonium chrysogenum (Cephalosporium acremonium). The biosynthetic pathways leading to both secondary metabolites start from the same three amino acid precursors and have the first two enzymatic reactions in common. Penicillin biosynthesis is catalyzed by three enzymes encoded by acvA (pcbAB), ipnA (pcbC), and aatA (penDE). The genes are organized into a cluster. In A. chrysogenum, in addition to acvA and ipnA, a second cluster contains the genes encoding enzymes that catalyze the reactions of the later steps of the cephalosporin pathway (cefEF and cefG). Within the last few years, several studies have indicated that the fungal β-lactam biosynthesis genes are controlled by a complex regulatory network, e.g., by the ambient pH, carbon source, and amino acids. A comparison with the regulatory mechanisms (regulatory proteins and DNA elements) involved in the regulation of genes of primary metabolism in lower eukaryotes is thus of great interest. This has already led to the elucidation of new regulatory mechanisms. Furthermore, such investigations have contributed to the elucidation of signals leading to the production of β-lactams and their physiological meaning for the producing fungi, and they can be expected to have a major impact on rational strain improvement programs. The knowledge of biosynthesis genes has already been used to produce new compounds.

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“…These species, along with Streptomyces clavuligerus ATCC 27064 (DSM 738), S. lactamdurans ATCC 27382 (NRRL 3802), Streptomyces jumonjinensis ATCC 29864, and Flavobacterium sp. strain SC 12.154 (Squibb) (30) were maintained on 2% Bacto agar (Difco) containing 1% glycerol (Fisher), 0.5% * Corresponding author. yeast extract (Difco), and 0.5% malt extract (Difco).…”

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Low-molecular-weight thiols in streptomycetes and their potential role as antioxidants

et al. 1993

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The intracellular low-molecular-weight thiols present in five gram-positive Streptomyces species and one Flavobacterium species were analyzed by high-performance liquid chromatography after fluorescence labeling with monobromobimane. Bacteria were chosen to include penicillin and cephalosporin 13-lactam producers and nonproducers. No significant amount of glutathione was found in any of the streptomycetes. Major intracellular thiols in all strains examined were cysteine, coenzyme A, sulfide, thiosulfate, and an unknown thiol designated U17. Those streptomycetes that make B-lactam antibiotics also produce significant amounts of 8-(L-aaminoadipyl)-L-cysteinyl-D-valine (ACV), a key intermediate in their biosynthesis. In Streptomyces clavuligerus, a potent producer of fl-lactams, the level of ACV was low during the early phase of growth and increased rapidly toward the end of exponential growth, paralleling that of antibiotic production. These and other observations indicate that ACV does not function as a protective thiol in streptomycetes. U17 may have this role since it was the major thiol in all streptomycetes and appeared to occur at levels about 10-fold higher than those of the other thiols measured, including ACV. Purification and amino acid analysis of U17 indicated that it contains cysteine and an unusual amine that is not one of the common amino acids. This thiol is identical to an unknown thiol found previously in Micrococcus roseus and Streptomyces griseus. A high level of ergothioneine was found in Streptomyces lactamdurans, and several unidentified thiols were detected in this and other streptomycetes.Numerous species of Streptomyces and one known species of Nocardia synthesize sulfur-containing penicillin, cephalosporin, and cephamycin antibiotics (2). Early investigators of the biosyntheses of these P-lactam compounds noticed the close structural relationship (28) between a key intermediate in the pathway, b-(L-oa-aminoadipyl)-L-cysteinyl-D-valine (ACV) and reduced glutathione, y-L-glutamyl-L-cysteinyl-glycine (GSH) (Fig. 1). GSH is the predominant thiol in eucaryotic organisms, in which it plays an important role in protecting cells against oxygen toxicity (17). It is found in most gram-negative bacteria but has not been detected in several major classes of gram-positive bacteria (13,14,26). Conflicting results were reported for the presence of GSH in streptomycetes. In one study, GSH could not be detected in cell extracts of Streptomyces griseus (14).In another study, low levels of GSH were found in Streptomyces lactamdurans (syn. Nocardia lactamdurans) (9). One purpose of the present study was to establish whether streptomycetes are able to produce GSH. Another was to determine whether ACV or other low-molecular-weight thiols play an antioxidant role in streptomycetes similar to that of GSH in other organisms. yeast extract (Difco), and 0.5% malt extract (Difco). Trypticase soy broth (TSB; BBL Becton Dickinson Microbiology Systems) was chosen for growth of these strains in liquid culture since it ...

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

Cited by 20 publications

References 47 publications

Molecular and Biochemical Heterogeneity of Class B Carbapenem-Hydrolyzing β-Lactamases in Chryseobacterium meningosepticum

Molecular and Biochemical Heterogeneity of Class B Carbapenem-Hydrolyzing β-Lactamases in Chryseobacterium meningosepticum

Molecular Regulation of β-Lactam Biosynthesis in Filamentous Fungi

Low-molecular-weight thiols in streptomycetes and their potential role as antioxidants

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