PENICILLIN AND CEPHALOSPORIN BIOSYNTHETIC GENES: Structure, Organization, Regulation, and Evolution

Aharonowitz, Yair; Cohen, Gerald; Martı́n, Juan F.

doi:10.1146/annurev.mi.46.100192.002333

Cited by 252 publications

(133 citation statements)

References 28 publications

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“…and then the cyclization of ACV to make isopenicillin N by the enzyme isopenicillin N synthase (IPNS) (3). ACV can exist in two states, either as the reduced monomeric form, which possesses a free cysteine thiol, or as the oxidized disulfide (bis-ACV) dimer.…”

Section: -(L-ct-aminoadipyl)-l-cysteinyl-d-valine (Acv) Synthetasementioning

confidence: 99%

The thioredoxin system of Penicillium chrysogenum and its possible role in penicillin biosynthesis

et al. 1994

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Penicillium chrysogenum is an important producer of penicillin antibiotics. A key step in their biosynthesis is the oxidative cyclization of 8-(L-01-aminoadipyl)-L-cysteinyl-D-valine (ACV) to isopenicillin N by the enzyme isopenicillin N synthase (IPNS). bis-ACV, the oxidized disulfide form of ACV is, however, not a substrate for IPNS. We report here the characterization of a broad-range disulfide reductase from P. chrysogenum that efficiently reduces bis-ACV to the thiol monomer. When coupled in vitro with IPNS, it converts bis-ACV to isopenicillin N and may therefore play a role in penicillin biosynthesis. The disulfide reductase consists of two protein components, a 72-kDa NADPH-dependent reductase, containing two identical subunits, and a 12-kDa general disulfide reductant. The latter reduces disulfide bonds in low-molecular-weight compounds and in proteins. The genes coding for the reductase system were cloned and sequenced. Both possess introns. A comparative analysis of their predicted amino acid sequences showed that the 12-kDa protein shares 26 to 600% sequence identity with thioredoxins and that the 36-kDa protein subunit shares 44 to 49%o sequence identity with the two known bacterial thioredoxin reductases. In addition, the P. chrysogenum NADPH-dependent reductase is able to accept thioredoxin as a substrate. These results establish that the P. chrysogenum broad-range disulfide reductase is a member of the thioredoxin family of oxidoreductases. This is the first example of the cloning of a eucaryotic thioredoxin reductase gene.Penicillins are sulfur-containing f-lactam compounds that are produced by certain filamentous fungi and industrially by Penicillium chrysogenum. The initial steps in the biosynthesis of these antibiotics involve the condensation of a-aminoadipic acid, cysteine, and valine to form a tripeptide by the enzyme coupled with IPNS, it quantitatively converts bis-ACV to isopenicillin N. Because ACV structurally resembles glutathione, y-glutamyl-cysteinyl-glycine, the most common intracellular low-molecular-weight (LMW) thiol, we originally supposed that this disulfide reductase might be related to glutathione reductase. However, we have now shown that streptomycetes lack both glutathione and glutathione reductase (2, 35). Also, a biochemical characterization of the S. clavuligerus disulfide reductase revealed that, in contrast to glutathione reductase, the S. clavuligerus disulfide reductase reduces a wide range of disulfides in LMW compounds and in proteins and is composed of two nonidentical polypeptides. The high-molecular-weight (HMW) component is a 70-kDa flavoprotein containing two identical subunits. In the presence of NADPH, it catalyzes the transfer of electrons to the LMW component. The latter is a heat-stable 12-kDa protein that is a general disulfide reductant. These properties of the S. clavuligerus broad-range disulfide reductase led us to propose that it belongs to the thioredoxin-thioredoxin reductase class of flavoprotein disulfide oxidoreductases. This proposal was r...

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Section: -(L-ct-aminoadipyl)-l-cysteinyl-d-valine (Acv) Synthetasementioning

confidence: 99%

The thioredoxin system of Penicillium chrysogenum and its possible role in penicillin biosynthesis

et al. 1994

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“…Because of the high GC content of the fungal orthologous genes we hypothesize that the gene was acquired by the fungal strains by horizontal gene transfer from streptomycetes in their common ecological niche. It is reasonable to suppose that these filamentous soil microorganisms eat each otherstreptomycetes possess batteries of chitinases that would help them to digest fungal walls, and many fungi make antibiotics active against bacterial cell wall biosynthesis, including β-lactams; and the fungal genes for β-lactam biosynthesis are themselves likely to have been acquired by lateral transfer [24]. Acquisition of the facC gene could 11 help filamentous fungal species in the competition with streptomycetes perhaps by increasing the rate of sporulation.…”

Section: Resultsmentioning

confidence: 99%

Novel and sensitive qPCR assays for the detection and identification of aspergillosis causing species

Paholcsek

Leiter

Markovics

et al. 2014

Acta Microbiologica Et Immunologica Hungarica

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Despite concerted efforts, diagnosis of aspergillosis is still a great challenge to clinical microbiology laboratories. Along with the requirement for high sensitivity and specificity, species-specific identification is important. We developed rapid, sensitive and species-specific qPCR assays using the TaqMan by Aspergillus fumigatus and Aspergillus terreus, which merits future diagnostic studies. The assays were sensitive enough to detect a few genomic equivalents in blood samples.

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“…Cephalosporin C belongs to the class of broad-spectrum antibiotics obtained from cultures of the filamentous fungus A. chrysogenum (114,115) . The cephalosporin C biosynthetic pathway begins with the non-ribosomal condensation of the three precursor amino acids l -α -aminoadipic acid, l -cysteine, and l -valine to form the tripeptide ACV (116) .…”

Section: Role Of Two Peroxisomal Membrane Transporters In Cephalospormentioning

confidence: 99%

Transport of substrates into peroxisomes: the paradigm of β-lactam biosynthetic intermediates

Martı́n

Garcı́a-Estrada

Ullán

2013

BioMolecular Concepts

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Peroxisomes are ubiquitous organelles that enclose catalases, fatty acid-oxidizing enzymes, and a variety of proteins involved in different cellular processes. Interestingly, the late enzymes involved in penicillin biosynthesis, and the isopenicillin N epimerization enzymes involved in cephalosporin biosynthesis are located inside peroxisomes in the producer fungi Penicillium chrysogenum and Acremonium chrysogenum. Peroxisome proteins are targeted to those organelles by peroxisomal targeting signals located at the C-terminus (PTS1) or near the N-terminal end (PTS2) of those proteins. Peroxisomal membrane proteins (PMPs) are largely recruited by the interaction with specific sequences in the Pex19 protein. The compartmentalization into peroxisomes of several steps of the biosynthesis of penicillin, cephalosporin, and other secondary metabolites raises the question of how the precursors and/or intermediates of the biosynthesis of β -lactam antibiotics are transported into peroxisomes and the mechanisms of secretion of the final products (penicillin or cephalosporin) from peroxisomes to the extracellular medium. Recent advances in peroxisome proteomics, immunoelectron microscopy, and fluorescence labeling have shown that the transport of these intermediates is mediated by membrane proteins of the major facilitator superfamily class (drug/H + antiporters) containing 12 transmembrane-spanning domains (TMS). In some cases, the transport of the substrates (e.g., fatty acids) or intermediates may be mediated by ATP-binding cassette (ABC) transporters. Knowledge on the transport and secretion mechanisms is of paramount importance to understand the complex mechanisms of cell differentiation and their crosstalk with the biosynthesis of different secondary metabolites that act as biochemical signals between the producer cells and also as communication signals with competing microorganisms (e.g., antimicrobial agents or plant elicitors).

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PENICILLIN AND CEPHALOSPORIN BIOSYNTHETIC GENES: Structure, Organization, Regulation, and Evolution

Cited by 252 publications

References 28 publications

The thioredoxin system of Penicillium chrysogenum and its possible role in penicillin biosynthesis

The thioredoxin system of Penicillium chrysogenum and its possible role in penicillin biosynthesis

Novel and sensitive qPCR assays for the detection and identification of aspergillosis causing species

Transport of substrates into peroxisomes: the paradigm of β-lactam biosynthetic intermediates

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