Metabolic plasticity for isoprenoid biosynthesis in bacteria

2013 ◽  
Vol 452 (1) ◽  
pp. 19-25 ◽  
Author(s):  
Jordi Pérez-Gil ◽  
Manuel Rodríguez-Concepción
Keyword(s):  
Mevalonate Pathway ◽  
Large Family ◽  
Isoprenoid Biosynthesis ◽  
Mep Pathway ◽  
Host Cells ◽  
Mva Pathway ◽  
Metabolic Intermediates ◽  
Metabolic Plasticity ◽  
New Antibiotics ◽  
Living Organisms

Isoprenoids are a large family of compounds synthesized by all free-living organisms. In most bacteria, the common precursors of all isoprenoids are produced by the MEP (methylerythritol 4-phosphate) pathway. The MEP pathway is absent from archaea, fungi and animals (including humans), which synthesize their isoprenoid precursors using the completely unrelated MVA (mevalonate) pathway. Because the MEP pathway is essential in most bacterial pathogens (as well as in the malaria parasites), it has been proposed as a promising new target for the development of novel anti-infective agents. However, bacteria show a remarkable plasticity for isoprenoid biosynthesis that should be taken into account when targeting this metabolic pathway for the development of new antibiotics. For example, a few bacteria use the MVA pathway instead of the MEP pathway, whereas others possess the two full pathways, and some parasitic strains lack both the MVA and the MEP pathways (probably because they obtain their isoprenoids from host cells). Moreover, alternative enzymes and metabolic intermediates to those of the canonical MVA or MEP pathways exist in some organisms. Recent work has also shown that resistance to a block of the first steps of the MEP pathway can easily be developed because several enzymes unrelated to isoprenoid biosynthesis can produce pathway intermediates upon spontaneous mutations. In the present review, we discuss the major advances in our knowledge of the biochemical toolbox exploited by bacteria to synthesize the universal precursors for their essential isoprenoids.

Properties and inhibition of the first two enzymes of the non-mevalonate pathway of isoprenoid biosynthesis

2000 ◽  
Vol 28 (6) ◽  
pp. 792-793 ◽  
Author(s):  
C. Mueller ◽  
J. Schwender ◽  
J. Zeidler ◽  
H. K. Lichtenthaler
Keyword(s):  
Escherichia Coli ◽  
Mevalonate Pathway ◽  
Isoprenoid Biosynthesis ◽  
Mep Pathway ◽  
Ph Optimum ◽  
Antibacterial Drugs ◽  
Chloroplast Stroma

Enzymes of the 1-deoxy-D-xylulose 5-phosphate/2-C-methylerythritol 4-phosphate (DOXP/MEP) pathway are targets for new herbicides and antibacterial drugs. Until now, no inhibitors for the DOXP synthase have been known of. We show that one of the breakdown products of the herbicide clomazone affects the DOXP synthase. One inhibitor of the non-mevalonate pathway, fosmidomycin, blocks the DOXP reductoisomerase (DXR) of plants and bacteria. The I50 values of plants are, however, higher than those found for the DXR of Escherichia coli. The DXR of plants, isolated from barley seedlings, shows a pH optimum of 8.1, which is typical for enzymes active in the chloroplast stroma.

scholarly journals Isoprenoid biosynthesis via 1-deoxy-D-xylulose 5-phosphate/2-C-methyl-D-erythritol 4-phosphate (DOXP/MEP) pathway.

2001 ◽  
Vol 48 (3) ◽  
pp. 663-672 ◽  
Author(s):  
M Wanke ◽  
K Skorupinska-Tudek ◽  
E Swiezewska
Keyword(s):  
Mevalonate Pathway ◽  
Higher Plants ◽  
Developmental State ◽  
Isoprenoid Biosynthesis ◽  
Mep Pathway ◽  
The Novel ◽  
Dimethylallyl Diphosphate ◽  
Parasite Plasmodium ◽  
Parasite Plasmodium Falciparum ◽  
Mixed Origin

Higher plants, several algae, bacteria, some strains of Streptomyces and possibly malaria parasite Plasmodium falciparum contain the novel, plastidic DOXP/MEP pathway for isoprenoid biosynthesis. This pathway, alternative with respect to the classical mevalonate pathway, starts with condensation of pyruvate and glyceraldehyde-3-phosphate which yields 1-deoxy-D-xylulose 5-phosphate (DOXP); the latter product can be converted to isopentenyl diphosphate (IPP) and eventually to isoprenoids or thiamine and pyridoxal. Subsequent reactions of this pathway involve transformation of DOXP to 2-C-methyl-D-erythritol 4-phosphate (MEP) which after condensation with CTP forms 4-diphosphocytidyl-2-amethyl-D-erythritol (CDP-ME). Then CDP-ME is phosphorylated to 4-diphosphocytidyl-2-amethyl-D-erythritol 2-phosphate (CDP-ME2P) and to 2-C-methyl-D-erythritol-2,4-cyclodiphosphate (ME-2,4cPP) which is the last known intermediate of the DOXP/MEP pathway. For- mation of IPP and dimethylallyl diphosphate (DMAPP) from ME-2,4cPP still requires clarification. This novel pathway appears to be involved in biosynthesis of carotenoids, phytol (side chain of chlorophylls), isoprene, mono-, di-, tetraterpenes and plastoquinone whereas the mevalonate pathway is responsible for formation of sterols, sesquiterpenes and triterpenes. Several isoprenoids were found to be of mixed origin suggesting that some exchange and/or cooperation exists between these two pathways of different biosynthetic origin. Contradictory results described below could indicate that these two pathways are operating under different physiological conditions of the cell and are dependent on the developmental state of plastids.

Non-mevalonate isoprenoid biosynthesis: enzymes, genes and inhibitors

2000 ◽  
Vol 28 (6) ◽  
pp. 785-789 ◽  
Author(s):  
H. K. Lichtenthaler
Keyword(s):  
Malaria Parasite ◽  
Pathogenic Bacteria ◽  
Mevalonate Pathway ◽  
Isoprenoid Biosynthesis ◽  
Mep Pathway ◽  
Isopentenyl Diphosphate ◽  
The Novel ◽  
Isoprenoid Pathway

The essential steps of the novel non-mevalonate pathway of isopentenyl diphosphate and isoprenoid biosynthesis in plants are described. The first five enzymes and genes of this 1-deoxy-D-xylulose 5-phosphate/2-C-methyl-D-erythritol 4-phosphate (DOXP/MEP) pathway are known. The herbicide fosmidomycin specifically blocks the second enzyme, the DOXP reductoisomerase. The DOXP/MEP pathway is also present in several pathogenic bacteria and the malaria parasite. Hence, all herbicides and inhibitors blocking this novel isoprenoid pathway in plants are also potential drugs against malaria and diseases caused by pathogenic bacteria.

scholarly journals GcpE Is Involved in the 2-C-Methyl-d-Erythritol 4-Phosphate Pathway of Isoprenoid Biosynthesis in Escherichia coli

2001 ◽  
Vol 183 (8) ◽  
pp. 2411-2416 ◽  
Author(s):  
Boran Altincicek ◽  
Ann-Kristin Kollas ◽  
Silke Sanderbrand ◽  
Jochen Wiesner ◽  
Martin Hintz ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Mevalonate Pathway ◽  
Genetically Engineered ◽  
Isoprenoid Biosynthesis ◽  
Mep Pathway ◽  
E Coli

ABSTRACT In a variety of organisms, including plants and several eubacteria, isoprenoids are synthesized by the mevalonate-independent 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway. Although different enzymes of this pathway have been described, the terminal biosynthetic steps of the MEP pathway have not been fully elucidated. In this work, we demonstrate that thegcpE gene of Escherichia coli is involved in this pathway. E. coli cells were genetically engineered to utilize exogenously provided mevalonate for isoprenoid biosynthesis by the mevalonate pathway. These cells were then deleted for the essential gcpE gene and were viable only if the medium was supplemented with mevalonate or the cells were complemented with an episomal copy of gcpE.

scholarly journals A Whole-Cell Phenotypic Screening Platform for Identifying Methylerythritol Phosphate Pathway-Selective Inhibitors as Novel Antibacterial Agents

2012 ◽  
Vol 56 (9) ◽  
pp. 4906-4913 ◽  
Author(s):  
Charles A. Testa ◽  
L. Jeffrey Johnson
Keyword(s):  
High Throughput Screening ◽  
Mep Pathway ◽  
Phenotypic Screening ◽  
Mva Pathway ◽  
Whole Cell ◽  
Content Type ◽  
Methylerythritol Phosphate ◽  
Serovar Typhimurium ◽  
Living Organisms ◽  
Screening Platform

ABSTRACTIsoprenoid biosynthesis is essential for survival of all living organisms. More than 50,000 unique isoprenoids occur naturally, with each constructed from two simple five-carbon precursors: isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Two pathways for the biosynthesis of IPP and DMAPP are found in nature. Humans exclusively use the mevalonate (MVA) pathway, while most bacteria, including all Gram-negative and many Gram-positive species, use the unrelated methylerythritol phosphate (MEP) pathway. Here we report the development of a novel, whole-cell phenotypic screening platform to identify compounds that selectively inhibit the MEP pathway. Strains ofSalmonella entericaserovar Typhimurium were engineered to have separately inducible MEP (native) and MVA (nonnative) pathways. These strains, RMC26 and CT31-7d, were then used to differentiate MVA pathway- and MEP pathway-specific perturbation. Compounds that inhibit MEP pathway-dependent bacterial growth but leave MVA-dependent growth unaffected represent MEP pathway-selective antibacterials. This screening platform offers three significant results. First, the compound is antibacterial and is therefore cell permeant, enabling access to the intracellular target. Second, the compound inhibits one or more MEP pathway enzymes. Third, the MVA pathway is unaffected, suggesting selectivity for targeting the bacterial versus host pathway. The cell lines also display increased sensitivity to two reported MEP pathway-specific inhibitors, further biasing the platform toward inhibitors selective for the MEP pathway. We demonstrate development of a robust, high-throughput screening platform that combines phenotypic and target-based screening that can identify MEP pathway-selective antibacterials simply by monitoring optical density as the readout for cell growth/inhibition.

scholarly journals The mevalonate pathway contributes to monoterpene production in peppermint

2020 ◽  
Author(s):  
Somnath Koley ◽  
Eva Grafahrend-Belau ◽  
Manish L. Raorane ◽  
Björn H. Junker
Keyword(s):  
Metabolic Flux ◽  
Mevalonate Pathway ◽  
Glandular Trichomes ◽  
Quantitative Description ◽  
Pentose Phosphate ◽  
Mep Pathway ◽  
Flux Analysis ◽  
Mva Pathway ◽  
Isoprenoid Metabolism ◽  
Monoterpene Biosynthesis

ABSTRACTPeppermint produces monoterpenes which are of great commercial value in different traditional and modern pharmaceutical and cosmetic industries. In the classical view, monoterpenes are synthesized via the plastidic 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway, while the cytosolic mevalonate (MVA) pathway produces sesquiterpenes. Interactions between both pathways have been documented in several other plant species, however, a quantitative understanding of the metabolic network involved in monoterpene biosynthesis is still lacking. Isotopic tracer analysis, steady state 13C metabolic flux analysis (MFA) and pathway inhibition studies were applied in this study to quantify metabolic fluxes of primary and isoprenoid metabolism of peppermint glandular trichomes (GT). Our results offer new insights into peppermint GT metabolism by confirming and quantifying the crosstalk between the two isoprenoid pathways towards monoterpene biosynthesis. In addition, a quantitative description of precursor pathways involved in isoprenoid metabolism is given. While glycolysis was shown to provide precursors for the MVA pathway, the oxidative bypass of glycolysis fueled the MEP pathway, indicating prominent roles for the oxidative branch of the pentose phosphate pathway and RuBisCO. This study reveals the potential of 13C-MFA to ascertain previously unquantified metabolic routes of the trichomes and thus advancing insights on metabolic engineering of this organ.

scholarly journals Cross-talk between the methylerythritol phosphate and mevalonic acid pathways of isoprenoid biosynthesis in poplar

2020 ◽  
Author(s):  
Hui Wei ◽  
Ali Movahedi ◽  
Chen Xu ◽  
Weibo Sun ◽  
Pu Wang ◽  
...  
Keyword(s):  
Cross Talk ◽  
Populus Trichocarpa ◽  
Mevalonic Acid ◽  
Isoprenoid Biosynthesis ◽  
Mep Pathway ◽  
Mva Pathway ◽  
Transcript Levels ◽  
Expression Levels ◽  
Methylerythritol Phosphate ◽  
Transgenic Poplars

Plants use two distinct isoprenoid biosynthesis pathways: the methylerythritol phosphate (MEP) pathway and mevalonic acid (MVA) pathway. 1-deoxy-D-xylulose5-phosphate synthase (DXS) and 1-deoxy-D-xylulose5-phosphate reductoisomerase (DXR) are the rate-limiting enzymes in the MEP pathway, and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) is a key regulatory enzyme in the MVA pathway. Previously, overexpression of Populus trichocarpa PtDXR in Nanlin 895 poplar was found to upregulate resistance to salt and drought stresses, and the transgenic poplars showed improved growth. In the present study, PtHMGR overexpressors (OEs) exhibited higher expression levels of DXS, DXR, 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase (HDS), and 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate reductase (HDR) and lower expression levels of 2-C-methyl-d-erythritol4-phosphate cytidylyltransferase (MCT), 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase (CMK), and 3-hydroxy-3-methylglutaryl-coenzyme A synthase (HMGS) than non-transgenic poplars (NT). However, the poplar PtDXR-OEs showed upregulated expression levels of MEP-related genes and downregulated expression of MVA-related genes. Moreover, overexpression of PtDXR and PtHMGR in poplars caused changes in MVA-derived trans-zeatin-riboside (TZR), isopentenyl adenosine (IPA), castasterone (CS) and 6-deoxocastasterone (DCS), as well as MEP-derived carotenoids, gibberellins (GAs), and abscisic acid (ABA). In PtHMGR-OEs, greater accumulation of geranyl diphosphate synthase (GPS) and geranyl pyrophosphate synthase (GPPS) transcript levels in the MEP pathway led to accumulation of MEP-derived isoprenoids, while upregulation of farnesyl diphosphate synthase (FPS) expression in the MVA pathway contributed to increased levels of MVA-derived isoprenoids. Similarly, in PtDXR-OEs, increased GPS and GPPS transcript levels in the MEP pathway boosted MEP-derived isoprenoid levels and changes in FPS expression affected MVA-derived isoprenoid yields. From these results, we can conclude that cross-talk exists between the MVA and MEP pathways.

scholarly journals Stable isotope and chemical inhibition analyses suggested the existence of a non-mevalonate-like pathway in the yeast Yarrowia lipolytica

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sivamoke Dissook ◽  
Tomohisa Kuzuyama ◽  
Yuri Nishimoto ◽  
Shigeru Kitani ◽  
Sastia Putri ◽  
...  
Keyword(s):  
Yarrowia Lipolytica ◽  
Oleaginous Yeast ◽  
Isoprenoid Biosynthesis ◽  
Mep Pathway ◽  
Authentic Standard ◽  
Chemical Inhibition ◽  
Chromatographic Peaks ◽  
Methyl Erythritol Phosphate ◽  
Limiting Condition ◽  
Yeast Yarrowia Lipolytica

AbstractMethyl erythritol phosphate (MEP) is the metabolite found in the MEP pathway for isoprenoid biosynthesis, which is known to be utilized by plants, algae, and bacteria. In this study, an unprecedented observation was found in the oleaginous yeast Yarrowia lipolytica, in which one of the chromatographic peaks was annotated as MEP when cultivated in the nitrogen limiting condition. This finding raised an interesting hypothesis of whether Y. lipolytica utilizes the MEP pathway for isoprenoid biosynthesis or not, because there is no report of yeast harboring the MEP pathway. Three independent approaches were used to investigate the existence of the MEP pathway in Y. lipolytica; the spiking of the authentic standard, the MEP pathway inhibitor, and the 13C labeling incorporation analysis. The study suggested that the mevalonate and MEP pathways co-exist in Y. lipolytica and the nitrogen limiting condition triggers the utilization of the MEP pathway in Y. lipolytica.

The Non-Mevalonate Isoprenoid Biosynthesis of Plants as a Test System for New Herbicides and Drugs against Pathogenic Bacteria and the Malaria Parasite

2000 ◽  
Vol 55 (5-6) ◽  
pp. 305-313 ◽  
Author(s):  
Hartmut K. Lichtenthaler ◽  
Johannes Zeidler ◽  
Jörg Schwender ◽  
Christian Müller
Keyword(s):  
Malaria Parasite ◽  
Pathogenic Bacteria ◽  
Higher Plants ◽  
Test System ◽  
Isoprenoid Biosynthesis ◽  
Mep Pathway ◽  
New Drugs ◽  
Security Measures ◽  
Isoprenoid Pathway ◽  
Parasite Plasmodium Falciparum

Higher plants and several photosynthetic algae contain the plastidic 1-deoxy-ᴅ-xylulose 5- phosphate / 2-C-methyl-ᴅ-erythritol 4-phosphate pathway (DOXP/MEP pathway) for isoprenoid biosynthesis. The first four enzymes and their genes are known of this novel pathway. All of the ca. 10 enzymes of this isoprenoid pathway are potential targets for new classes of herbicides. Since the DOXP/MEP pathway also occurs in several pathogenic bacteria, such as Mycobacterium tuberculosis, and in the malaria parasite Plasmodium falciparum, all inhibitors and potential herbicides of the DOXP/MEP pathway in plants are also potential drugs against pathogenic bacteria and the malaria parasite. Plants with their easily to handle DOXP/MEP-pathway are thus very suitable test-systems also for new drugs against pathogenic bacteria and the malaria parasite as no particular security measures are required. In fact, the antibiotic herbicide fosmidomycin specifically inhibited not only the DOXP reductoisomerase in plants, but also that in bacteria and in the parasite P. falciparum, and cures malaria-infected mice. This is the first successful application of a herbicide of the novel isoprenoid pathway as a possible drug against malaria.

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