Sequencing and overexpression of the <i>Escherichia coli aroE</i> gene encoding shikimate dehydrogenase

Anton, Ian A.; Coggins, John R.

doi:10.1042/bj2490319

Cited by 56 publications

(40 citation statements)

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“…1). The NADP-dependent shikimate-specific enzyme is involved in the biosynthetic role (1). On the other hand, it is likely that the enzyme involved in the catabolic role acts on both quinate and shikimate as substrates and prefers NAD as a coenzyme in fungi and gram-positive bacteria, although genetic characterization of the bacterial enzymes is obscure (3,12,14).…”

Section: Vol 75 2009mentioning

confidence: 99%

“…Unrooted phylogenetic tree showing relationships among bacterial shikimate dehydrogenase family proteins and the homologues in C. glutamicum. The proteins analyzed were E. coli AroE and YdiB (AroE-Ec and YdiB-Ec) (1,24), H. influenzae AroE and SdhL (AroE-Hi and SdhL-Hi) (36,47), M. tuberculosis shikimate dehydrogenase (SDH-Mt) (10), Amycolatopsis mediterranei RifI (RifI-Am) (48), and C. glutamicum QsuD, CgR_1216, and CgR_1677, encoded by qsuD, cgR_1216, and cgR_1677, respectively. The amino acid sequences were aligned using the CLUSTAL W program (40), and the tree was constructed using the neighbor-joining method (31).…”

Section: Vol 75 2009mentioning

confidence: 99%

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Regulation of Expression of Genes Involved in Quinate and Shikimate Utilization in Corynebacterium glutamicum

Teramoto

Inui

Yukawa

2009

Appl Environ Microbiol

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The utilization of the hydroaromatic compounds quinate and shikimate by Corynebacterium glutamicum was investigated. C. glutamicum grew well with either quinate or shikimate as the sole carbon source. The disruption of qsuD, encoding quinate/shikimate dehydrogenase, completely suppressed growth with either substrate but did not affect growth with glucose, indicating that the enzyme encoded by qsuD catalyzes the first step of the catabolism of quinate/shikimate but is not involved in the shikimate pathway required for the biosynthesis of various aromatic compounds. On the chromosome of C. glutamicum, the qsuD gene is located in a gene cluster also containing qsuA, qsuB, and qsuC genes, which are probably involved in the quinate/shikimate utilization pathway to form protocatechuate. Reverse transcriptase PCR analyses revealed that the expression of the qsuABCD genes was markedly induced during growth with either quinate or shikimate relative to expression during growth with glucose. The induction level by shikimate was significantly decreased by the disruption of qsuR, which is located immediately upstream of qsuA in the opposite direction and encodes a LysR-type transcriptional regulator, suggesting that QsuR acts as an activator of the qsuABCD genes. The high level of induction of qsuABCD genes by shikimate was still observed in the presence of glucose, and simultaneous consumption of glucose and shikimate during growth was observed.The abundant plant products quinate and shikimate are utilized as carbon and/or energy sources by various microorganisms. The quinate utilization pathways in the filamentous fungi Neurospora crassa and Aspergillus nidulans have been well studied ( Fig. 1) (12, 14). Quinate is converted to protocatechuate via three enzymatic reactions, catalyzed by quinate dehydrogenase, dehydroquinate dehydratase, and dehydroshikimate dehydratase. Subsequently, protocatechuate is metabolized through the ␤-ketoadipate pathway. The expression of these enzymes is induced during growth on quinate and is subject to carbon catabolite repression (12)(13)(14). The first enzyme for the utilization of quinate, quinate dehydrogenase, also converts shikimate to dehydroshikimate. Dehydroquinate and dehydroshikimate are also intermediates of the shikimate pathway, leading to branched pathways of biosynthesis of various aromatic amino acids, vitamins, and quinones. The biosynthetic reactions of dehydroquinate dehydratase and shikimate dehydrogenase are the same as the quinate/shikimate catabolic reactions. However, separately from the inducible catabolic enzymes, a constitutive pentafunctional enzyme containing the dehydroquinate dehydratase and shikimate dehydrogenase activities is involved in the shikimate biosynthetic pathway (12,14). The pentafunctional enzyme contains regions with amino acid sequence similarity to five shikimate pathway enzymes encoded by separate genes in bacteria, implying multiple gene fusion during evolution.Among gram-negative bacteria, Acinetobacter species utilize quinate and shikimate ...

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Section: Vol 75 2009mentioning

confidence: 99%

Section: Vol 75 2009mentioning

confidence: 99%

Regulation of Expression of Genes Involved in Quinate and Shikimate Utilization in Corynebacterium glutamicum

Teramoto

Inui

Yukawa

2009

Appl Environ Microbiol

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“…nidufans (Anarom), S. cerevisiae (Scarom), and P. carinii (Pcarom) AROM enzymes and also a composite of the E. cofi monofunctional enzymes which have been arranged in the following order: DHQ synthase (residues 1-363) ; EPSP synthase (364-789) (Duncan et af., 1984); shikimate kinase (790-963) (Millar e f al., 1986); DHQ dehydratase (966121 5 ) and shikimate dehydrogenase (1216-1487) (Anton & Coggins, 1988). The sequence of DHQ dehydratase has been amended from that originally published (Duncan er af., 1986) following the corrections specified by Chaudhuri et af.…”

Section: Fig 3 Volfowing Three Pages)mentioning

confidence: 99%

The cloning and characterization of the arom gene of Pneumocystis carinii

Banerji

Wakefield²,

Allen³

et al. 1993

Journal of General Microbiology

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The avom gene, encoding a single polypeptide that catalyses five consecutive steps of the pre-chorismate aromatic amino acid biosynthetic pathway, has been cloned from the opportunistic pathogen Pneumocystis cavinii. There is a single open reading frame of 4788 bp which includes an intron of 45 bp that does not introduce a stop codon into the sequence. Thus, the derived amino acid sequence consists of 1581 residues, which is highly homologous to all fungal AROM proteins studied to date. These data support the view that P. cavinii is a fungus and imply that its aromatic amino acid biosynthesis is conventionally organized.

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“…Recently it has been shown, by site-directed mutagenesis of the A. nidulans AROM locus, that some ofthe arom polypeptide (including the bDHQase domain) can fold and function in Escherichia coli as monofunctional enzymes [24]. In addition, a study of protein sequences deduced from DNA sequence data has shown that the quinate dehydrogenase enzyme of the quinic acid utilization pathway and the shikimate dehydrogenase enzyme of the shikimate pathway have evolved from a common ancestral sequence [5,19]. Conversely, the comparison of bDHQase and cDHQase enzyme sequences has led to the proposal that the two enzymes arose by convergent evolution [23,25].…”

Section: Introductionmentioning

confidence: 99%

“…Lamb et al [12] have shown that the common metabolites 3-dehydroquinate and dehydroshikimate leak from the arom protein at a rate comparable with the flux of metabolites through the shikimate pathway and can be utilized by the enzymes of both pathways. A similar fused polypeptide is also present in Neurospora crassa [13][14][15] and Saccharomyces cerevisiae [16] but, in prokaryotes, steps two to six in the shikimate pathway are catalysed by distinct monofunctional enzymes [17][18][19][20][21] that are constitutively expressed and are not required to aggregate for activity. Evidence from DNA sequence analysis has strongly suggested that the complex AROM locus arose by multiple gene fusions to generate the pentafunctional arom polypeptide [22,23].…”

Section: Introductionmentioning

confidence: 99%

A comparison of the enzymological and biophysical properties of two distinct classes of dehydroquinase enzymes

1992

Biochemical Journal

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This paper compares the biophysical and mechanistic properties of a typical type I dehydroquinase (DHQase), from the biosynthetic shikimate pathway of Escherichia coli, and a typical type II DHQase, from the quinate pathway of Aspergillus nidulans. C.d. shows that the two proteins have different secondary-structure compositions; the type I enzyme contains approx. 50 % a-helix while the type II enzyme contains approx. 750 a-helix. The stability of the two types of DHQase was compared by denaturant-induced unfolding, as monitored by c.d., and by differential scanning calorimetry. The type II enzyme unfolds at concentrations of denaturant 4-fold greater than the type I and through a series of discrete transitions, while the type I enzyme unfolds in a single transition. These differences in conformational stability were also evident from the calorimetric experiments which show that type I DHQase unfolds as a single co-operative dimer at 57°C whereas the type II enzyme unfolds above 82°C and through a series of transitions suggesting higher orders of structure than that seen for the type I enzyme. Sedimentation and Mr analysis of both proteins by analytical ultracentrifugation is consistent with the unfolding data. The type I DHQase exists predominantly as a dimer with Mr = 46000 + 2000 (a weighted average affected by the presence of monomer) and has a sedimentation coefficient s520 = 4.12 (+0.08)-S whereas the type II enzyme is a dodecamer, weight-average Mr = 190000 + 10000 and has a sedimentation coefficient, so = 9.96 (+ 0.21) S. Although both enzymes have reactive histidine residues in the active site and can be inactivated by diethyl pyrocarbonate, the possibility that these structurally dissimilar enzymes catalyse the same dehydration reaction by the same catalytic mechanism is deemed unlikely by three criteria: (1) they have very different pH/logkcat profiles and pH optima; (2) imine intermediates, which are known to play a central role in the mechanism of type I enzymes, could not be detected (by borohydride reduction) in the type II enzyme; (3) unlike Schiff's base-forming type I enzymes, there are no conserved lysine residues in type II amino acid sequences.

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Sequencing and overexpression of the Escherichia coli aroE gene encoding shikimate dehydrogenase

Cited by 56 publications

References 50 publications

Regulation of Expression of Genes Involved in Quinate and Shikimate Utilization in Corynebacterium glutamicum

Regulation of Expression of Genes Involved in Quinate and Shikimate Utilization in Corynebacterium glutamicum

The cloning and characterization of the arom gene of Pneumocystis carinii

A comparison of the enzymological and biophysical properties of two distinct classes of dehydroquinase enzymes

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