N-Pyridyl-aminomethylene-bisphosphonic Acids Inhibit the First Enzyme in the Shikimate Pathway, 3-Deoxy-D-arabino- heptulosonate-7-phosphate Synthase

1996 ◽  
Vol 55 (3) ◽  
pp. 180-188 ◽  
Author(s):  
Giuseppe Forlani ◽  
Barbara Lejczak ◽  
Pawel Kafarski
Keyword(s):  
Shikimate Pathway ◽  
Bisphosphonic Acids ◽  
Phosphate Synthase

Differential expression of tomato (Lycopersicon esculentum L.) genes encoding shikimate pathway isoenzymes. I. 3-Deoxy-D-arabino-heptulosonate 7-phosphate synthase

1993 ◽  
Vol 23 (4) ◽  
pp. 697-706 ◽  
Author(s):  
J�rn G�rlach ◽  
Andreas Beck ◽  
John M. Henstrand ◽  
Avtar K. Handa ◽  
Klaus M. Herrmann ◽  
...  
Keyword(s):  
Lycopersicon Esculentum ◽  
Differential Expression ◽  
Shikimate Pathway ◽  
Genes Encoding ◽  
Phosphate Synthase

scholarly journals The Glyphosate Target Enzyme 5-Enolpyruvyl Shikimate 3-Phosphate Synthase (EPSPS) Contains Several EPSPS-Associated Domains in Fungi

Proceedings ◽  
2020 ◽  
Vol 76 (1) ◽  
pp. 6
Author(s):  
Tuomas Tall ◽  
Pere Puigbò
Keyword(s):  
Amino Acids ◽  
Shikimate Pathway ◽  
Aromatic Amino Acids ◽  
Comprehensive Analysis ◽  
Single Domain ◽  
Differential Impact ◽  
Herbicide Glyphosate ◽  
Target Enzyme ◽  
Phosphate Synthase

5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) is the central enzyme of the shikimate pathway to synthesize three aromatic amino acids in fungi, plants and prokaryotes. This enzyme is the target of the herbicide glyphosate. In most plants and prokaryotes, the EPSPS protein is constituted by a single domain, whereas in fungi, it contains several EPSPS-associated domains. Here, we perform a comprehensive analysis of 390 EPSPS proteins of fungi to determine the distribution and the evolution of the EPSPS-associated domains. The results of this study will be useful to determine the potential differential impact of glyphosate on alternative domain architectures in fungi.

scholarly journals Differential expression of 5-enol-pyruvyl-shikimate-3-phosphate synthase isoforms in elicitor-treated, cultured maize cells

2002 ◽  
Vol 29 (12) ◽  
pp. 1483 ◽  
Author(s):  
Giuseppe Forlani
Keyword(s):  
Phenylalanine Ammonia Lyase ◽  
Zea Mays L ◽  
Cultured Cells ◽  
Amino Acid Content ◽  
Shikimate Pathway ◽  
Activity Levels ◽  
Sharp Rise ◽  
Enzyme Form ◽  
Genes Encoding ◽  
Phosphate Synthase

The expression of two 5-enol-pyruvyl-shikimate-3-phosphate synthase (EC 2.5.1.19) isoforms was investigated in Zea mays L. suspension-cultured cells following exposure to a fungal elicitor. Activity levels of isozyme II specifically increased soon after treatment, in strict connection with induction of phenylalanine ammonia-lyase (PAL) and attainment of a new free-phenylalanine homeostasis at a higher concentration. However, a few days later, activity of the other enzyme form was also significantly enhanced, concomitant with a sharp rise in overall amino acid content, a further increase in PAL level and a resumption of cell lysis. Besides strengthening the hypothesis that an entire set of genes encoding for shikimate pathway enzymes (whose expression is specifically involved in plant dynamic defence) may exist, a general change in the levels of several amino acids seems to point towards a reprogramming of their metabolism in elicited cells.

The aromatic amino acid pathway branches at L-arogenate in Euglena gracilis

1981 ◽  
Vol 1 (5) ◽  
pp. 426-438
Author(s):  
G S Byng ◽  
R J Whitaker ◽  
C L Shapiro ◽  
R A Jensen
Keyword(s):  
Amino Acid ◽  
Euglena Gracilis ◽  
Nicotinamide Adenine Dinucleotide ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Shikimate Pathway ◽  
Exchange Chromatography ◽  
Chorismate Mutase ◽  
Nicotinamide Adenine ◽  
Acid Pathway ◽  
Phosphate Synthase

The recently characterized amino acid L-arogenate (Zamir et al., J. Am. Chem. Soc. 102:4499-4504, 1980) may be a precursor of either L-phenylalanine or L-tyrosine in nature. Euglena gracilis is the first example of an organism that uses L-arogenate as the sole precursor of both L-tyrosine and L-phenylalanine, thereby creating a pathway in which L-arogenate rather than prephenate becomes the metabolic branch point. E. gracilis ATCC 12796 was cultured in the light under myxotrophic conditions and harvested in late exponential phase before extract preparation for enzymological assays. Arogenate dehydrogenase was dependent upon nicotinamide adenine dinucleotide phosphate for activity. L-Tyrosine inhibited activity effectively with kinetics that were competitive with respect to L-arogenate and noncompetitive with respect to nicotinamide adenine dinucleotide phosphate. The possible inhibition of arogenate dehydratase by L-phenylalanine has not yet been determined. Beyond the latter uncertainty, the overall regulation of aromatic biosynthesis was studied through the characterization of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase and chorismate mutase. 3-Deoxy-D-arabino-heptulosonate 7-phosphate synthase was subject to noncompetitive inhibition by L-tyrosine with respect to either of the two substrates. Chorismate mutase was feedback inhibited with equal effectiveness by either L-tyrosine or L-phenylalanine. L-Tryptophan activated activity of chorismate mutase, a pH-dependent effect in which increased activation was dramatic above pH 7.8 L-Arogenate did not affect activity of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase or of chorismate mutase. Four species of prephenate aminotransferase activity were separated after ion-exchange chromatography. One aminotransferase exhibited a narrow range of substrate specificity, recognizing only the combination of L-glutamate with prephenate, phenylpyruvate, or 4-hydroxyphenylpyruvate. Possible natural relationships between Euglena spp. and fungi previously considered in the literature are discussed in terms of data currently available to define enzymological variation in the shikimate pathway.

scholarly journals The aromatic amino acid pathway branches at L-arogenate in Euglena gracilis.

1981 ◽  
Vol 1 (5) ◽  
pp. 426-438 ◽  
Author(s):  
G S Byng ◽  
R J Whitaker ◽  
C L Shapiro ◽  
R A Jensen
Keyword(s):  
Amino Acid ◽  
Euglena Gracilis ◽  
Nicotinamide Adenine Dinucleotide ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Shikimate Pathway ◽  
Exchange Chromatography ◽  
Chorismate Mutase ◽  
Nicotinamide Adenine ◽  
Acid Pathway ◽  
Phosphate Synthase

The recently characterized amino acid L-arogenate (Zamir et al., J. Am. Chem. Soc. 102:4499-4504, 1980) may be a precursor of either L-phenylalanine or L-tyrosine in nature. Euglena gracilis is the first example of an organism that uses L-arogenate as the sole precursor of both L-tyrosine and L-phenylalanine, thereby creating a pathway in which L-arogenate rather than prephenate becomes the metabolic branch point. E. gracilis ATCC 12796 was cultured in the light under myxotrophic conditions and harvested in late exponential phase before extract preparation for enzymological assays. Arogenate dehydrogenase was dependent upon nicotinamide adenine dinucleotide phosphate for activity. L-Tyrosine inhibited activity effectively with kinetics that were competitive with respect to L-arogenate and noncompetitive with respect to nicotinamide adenine dinucleotide phosphate. The possible inhibition of arogenate dehydratase by L-phenylalanine has not yet been determined. Beyond the latter uncertainty, the overall regulation of aromatic biosynthesis was studied through the characterization of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase and chorismate mutase. 3-Deoxy-D-arabino-heptulosonate 7-phosphate synthase was subject to noncompetitive inhibition by L-tyrosine with respect to either of the two substrates. Chorismate mutase was feedback inhibited with equal effectiveness by either L-tyrosine or L-phenylalanine. L-Tryptophan activated activity of chorismate mutase, a pH-dependent effect in which increased activation was dramatic above pH 7.8 L-Arogenate did not affect activity of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase or of chorismate mutase. Four species of prephenate aminotransferase activity were separated after ion-exchange chromatography. One aminotransferase exhibited a narrow range of substrate specificity, recognizing only the combination of L-glutamate with prephenate, phenylpyruvate, or 4-hydroxyphenylpyruvate. Possible natural relationships between Euglena spp. and fungi previously considered in the literature are discussed in terms of data currently available to define enzymological variation in the shikimate pathway.

scholarly journals Bacillus subtilis 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase revisited: resolution of two long-standing enigmas

2005 ◽  
Vol 390 (2) ◽  
pp. 583-590 ◽  
Author(s):  
Jing Wu ◽  
Galina Ya. Sheflyan ◽  
Ronald W. Woodard
Keyword(s):  
Bacillus Subtilis ◽  
Dipicolinic Acid ◽  
Shikimate Pathway ◽  
Chorismate Mutase ◽  
Highly Active ◽  
Bifunctional Enzymes ◽  
Iron And Zinc ◽  
Bivalent Metal Ions ◽  
Structure In Solution ◽  
Phosphate Synthase

The mono/bifunctional and metallo/non-metallo properties of Bacillus subtilis DAHPS (3-deoxy-D-arabino-heptulosonate 7-phosphate synthase) have been controversial for several decades. The present study investigated the DAHPSs from both the B. subtilis parent Marburg strain and the derivative strain 168 in detail and clarified the above two long-standing questions. The DAHPSs from the parent and the derivative 168 strains have identical sequence and are both bifunctional enzymes with a CM (chorismate mutase) activity and a DAHPS activity. The parent strain expresses a second independent monofunctional CM, encoded by aroH, that is highly active, while the 168 strain expresses an aroH containing a single residue mutation (A112V) that is significantly less active thus leading to previous confusion regarding the mono/bifunctionality of DAHPS. Metal analysis showed that B. subtilis DAHPS as isolated contained iron and zinc and is inactivated by dipicolinic acid; the inactive apoenzyme can be reactivated by bivalent metal ions, indicating that the enzyme is a metalloenzyme. The enzyme-bound metal is insensitive to EDTA treatment, leading to the previous conclusion that this DAHPS does not require a metal. The enzyme displays a homotetrameric structure in solution and appears to follow Michaelis–Menten kinetics with KmPEP=139±11.4 μM for phosphoenolpyruvate, KmE4P=1760±110 μM for D-erythrose 4-phosphate, kcat=4.6±0.1 s−1 for DAHPS activity and Kmchorismate=850±97 μM, kcat=0.41±0.01 s−1 for CM activity. B. subtilis DAHPS is inhibited by the Shikimate pathway intermediates prephenate and chorismate.

scholarly journals Classification of the glyphosate target enzyme (5-enolpyruvylshikimate-3-phosphate synthase)

2020 ◽  
Author(s):  
Lyydia Leino ◽  
Tuomas Tall ◽  
Marjo Helander ◽  
Irma Saloniemi ◽  
Kari Saikkonen ◽  
...  
Keyword(s):  
Gut Microbiome ◽  
Shikimate Pathway ◽  
Aromatic Amino Acids ◽  
Research Field ◽  
Human Gut ◽  
Human Gut Microbiome ◽  
The Core ◽  
Key Enzyme ◽  
Phosphate Synthase

ABSTRACTGlyphosate is the most common broad-spectrum herbicide. It targets the key enzyme of the shikimate pathway, 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), which synthesizes three essential aromatic amino acids (phenylalanine, tyrosine and tryptophan) in plants. Because the shikimate pathway is also found in many prokaryotes and fungi, the widespread use of glyphosate may have unsuspected impacts on the diversity and composition of microbial communities, including the human gut microbiome. Here, we introduce the first bioinformatics method to assess the potential sensitivity of organisms to glyphosate based on the type of EPSPS enzyme. We have precomputed a dataset of EPSPS sequences from thousands of species that will be an invaluable resource to advancing the research field. This novel methodology can classify sequences from >90% of eukaryotes and >80% of prokaryotes. A conservative estimate from our results shows that 54% of species in the core human gut microbiome are sensitive to glyphosate.

scholarly journals Molecular cloning and metabolomic characterization of the 5-enolpyruvylshikimate-3-phosphate synthase gene from Baphicacanthus cusia

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Jian Yu ◽  
Yihan Zhang ◽  
Shuju Ning ◽  
Qi Ye ◽  
Hexin Tan ◽  
...  
Keyword(s):  
Enzyme Assay ◽  
Shikimate Pathway ◽  
Pcr Analysis ◽  
Gene Encoding ◽  
High Accumulation ◽  
Antiviral Compounds ◽  
Practical Implications ◽  
Putative Mechanism ◽  
Phosphate Synthase

Abstract Background Indigo alkaloids, such as indigo, indirubin and its derivatives, have been identified as effective antiviral compounds in Baphicacanthus cusia. Evidence suggests that the biosynthesis of indigo alkaloids in plants occurs via the shikimate pathway. The enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) is involved in plant metabolism; however, its underlying putative mechanism of regulating the production of indigo alkaloids is currently unknown. Results One gene encoding EPSPS was isolated from B. cusia. Quantitative real-time PCR analysis revealed that BcEPSPS was expressed at the highest level in the stem and upregulated by methyl jasmonate (MeJA), salicylic acid (SA) and abscisic acid (ABA) treatment. The results of subcellular localization indicated that BcEPSPS is mainly expressed in both the plastids and cytosol, which has not been previously reported. An enzyme assay revealed that the heterogeneously expressed BcEPSPS protein catalysed the generation of 5-enolpyruvyl shikimate-3-phosphate. The overexpression of BcEPSPS in Isatis indigotica hairy roots resulted in the high accumulation of indigo alkaloids, such as indigo, secologanin, indole and isorhamnetin. Conclusions The function of BcEPSPS in catalysing the production of EPSP and regulating indigo alkaloid biosynthesis was revealed, which provided a distinct view of plant metabolic engineering. Our findings have practical implications for understanding the effect of BcEPSPS on active compound biosynthesis in B. cusia.

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