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Geranyl pyrophosphate

Geranyl pyrophosphate (GPP), also known as geranyl diphosphate, is an organic compound with the molecular formula C₁₀H₂₀O₇P₂, serving as a key intermediate in the mevalonate and methylerythritol phosphate (MEP) pathways of isoprenoid biosynthesis. It consists of a 10-carbon geranyl chain attached to a pyrophosphate group, formed through the head-to-tail condensation of dimethylallyl pyrophosphate (DMAPP) and isopentenyl pyrophosphate (IPP), a reaction catalyzed by geranyl pyrophosphate synthase enzymes.^[1]^[2] In cellular metabolism, GPP functions primarily as a precursor for the synthesis of monoterpenes, which are volatile compounds essential for plant defense, pollination, and aroma, as well as for the elongation to longer prenyl pyrophosphates such as farnesyl pyrophosphate (FPP) and geranylgeranyl pyrophosphate (GGPP), which support sesquiterpene, diterpene, and sterol production across organisms.^[3] Its dysregulation is implicated in metabolic disorders, and GPP analogs are explored in pharmacology for inhibiting enzymes like farnesyl pyrophosphate synthase to target cholesterol biosynthesis or cancer-related prenylation.^[1] In plants, compartmentalization of GPP production in plastids versus cytosol directs its flux toward specific terpenoid classes, highlighting its central role in metabolic diversity.^[2]

Chemical Identity

Molecular Structure

Geranyl pyrophosphate (GPP) is the pyrophosphate ester of geraniol, consisting of a branched 10-carbon isoprenoid chain esterified at the primary alcohol position with a diphosphate group. Its molecular formula is C₁₀H₁₇O₇P₂, corresponding to the trianionic form prevalent in biological contexts, with a molar mass of 311.19 g/mol. The IUPAC name is (2E)-3,7-dimethylocta-2,6-dien-1-yl trihydrogen diphosphate, reflecting the specific stereochemistry of the double bond at C2–C3. The structure features a linear chain of eight carbons with methyl substituents at C3 and C7, a trans (E) double bond between C2–C3, a double bond between C6–C7, and the pyrophosphate moiety (-O-P(O)(OH)-O-P(O)(OH)₂) linked via an oxygen to C1, forming an allylic system.^[4] The geranyl chain can be described as -CH₂(1)-CH(2)=C(3)(CH₃)-CH₂(4)-CH₂(5)-CH(6)=C(7)(CH₃)-CH₃(8), with the pyrophosphate attached to C1. The allylic carbon at C1 is -CH₂-OP₂O₆H₃, adjacent to the C2=C3 double bond, which enhances leaving group potential of the diphosphate during ionization.^[4] The E-configuration at the C2–C3 double bond establishes the trans geometry essential for the spatial arrangement in extended terpenoid chains during biosynthesis. This stereochemistry distinguishes GPP from its cis isomer neryl pyrophosphate and supports efficient chain elongation in isoprenoid assembly.^[4]

Nomenclature and Properties

Geranyl pyrophosphate, commonly abbreviated as GPP, is systematically named (2E)-3,7-dimethylocta-2,6-dien-1-yl diphosphate. It is also referred to as geranyl diphosphate (GDP) or trans-geranyl pyrophosphate, reflecting its role as the pyrophosphate ester of the monoterpenoid alcohol geraniol.^[5] These names highlight the trans configuration at the C2-C3 double bond and the C10 isoprenoid chain length.^[6] The compound typically exists as salts, such as the lithium, ammonium, or triammonium forms, which appear as colorless solids or solutions. These salts exhibit limited solubility in water (slightly soluble, with concentrations up to >5 mg/mL in aqueous buffers), but they are more readily soluble in polar organic solvents like methanol.^[7] The molecular formula is C₁₀H₂₀O₇P₂, with a molecular weight of 314.21 g/mol for the free acid form. Chemically, geranyl pyrophosphate contains a labile pyrophosphate anhydride bond (P-O-P linkage) that stores high energy (approximately -7.3 kcal/mol under standard conditions), rendering it susceptible to nucleophilic attack and cleavage.^[8] This bond, combined with the allylic position of the geranyl chain, facilitates reactivity as an electrophilic substrate in substitution reactions, where the diphosphate serves as an excellent leaving group.^[9] GPP demonstrates instability under acidic or basic conditions, undergoing hydrolysis to yield geraniol and inorganic phosphate via C-O bond scission.^[9] In neutral aqueous environments, it can slowly degrade through elimination pathways, forming geraniol or premyrcene-like products, particularly when trace metal ions like Mn²⁺ are present to catalyze the process.^[8] Storage as dry salts at low temperatures (-20°C) minimizes degradation, though exposure to moisture or pH extremes accelerates breakdown.^[7] Identification of GPP often relies on spectroscopic techniques, including ¹H NMR showing characteristic signals for the trans double bond (δ ≈ 5.4 ppm, J ≈ 7 Hz) and methyl groups (δ ≈ 1.6-1.7 ppm), alongside ³¹P NMR peaks around -10 to -12 ppm for the diphosphate moiety.^[10] Infrared (IR) spectroscopy reveals strong absorptions for P-O stretches (≈ 1100-1200 cm⁻¹) and C=C bonds (≈ 1650 cm⁻¹).^[11]

Biosynthesis

Enzymatic Formation

Geranyl pyrophosphate (GPP) is synthesized through the head-to-tail condensation of dimethylallyl pyrophosphate (DMAPP) and isopentenyl pyrophosphate (IPP), the universal C5 isoprenoid precursors, to form the C10 product GPP and inorganic pyrophosphate (PPi).^[12] This reaction is catalyzed by geranyl pyrophosphate synthase (GPPS; EC 2.5.1.1), a member of the prenyltransferase family that exhibits strict chain-length specificity for C10 products in most cases.^[13] The overall reaction can be represented as:

\text{DMAPP} + \text{IPP} \rightarrow \text{GPP} + \text{PP}_\text{i}

^[12] The catalytic mechanism of GPPS follows a stepwise ionization-condensation-elimination pathway. Initially, the enzyme facilitates the ionization of DMAPP by dissociation of its pyrophosphate group, generating a resonance-stabilized allylic carbocation intermediate coordinated by Mg²⁺ ions.^[13] Subsequently, the C4 carbon of IPP's double bond performs a nucleophilic attack on the carbocation at the C1 position of DMAPP in an Sₙ1-like fashion, forming a new C-C bond.^[13] The process concludes with the stereospecific abstraction of the pro-R hydrogen from the C2 position of the IPP-derived moiety, yielding the trans-configured double bond in GPP and restoring the enzyme for further catalysis.^[13] The active site of GPPS features two conserved aspartate-rich motifs, typically DDxxD, which bind two Mg²⁺ ions to coordinate the pyrophosphate moieties of DMAPP and IPP.^[13] Critical residues, such as Asp-83, Asp-84, Asp-89, Arg-94, Arg-95, and Lys-44 in the large subunit of mint (Mentha piperita) GPPS, stabilize the carbocation and facilitate proton transfer.^[14] Chain-length specificity is enforced by a hydrophobic pocket lined with bulky residues like Phe-112 and Phe-113 in mint GPPS, which restrict further IPP addition beyond C10.^[13] GPPS variants differ across organisms: monofunctional forms produce only GPP, while bifunctional enzymes can elongate to farnesyl pyrophosphate (FPP).^[15] In plants like mint (Mentha piperita), GPPS often assembles as a heterotetramer (LSU·SSU)₂, where the large subunit (LSU) provides catalytic activity and the small subunit (SSU) enhances specificity by forming regulatory loops that limit product chain length.^[14] In contrast, GPPS in some plants, such as Norway spruce (Picea abies), may function as homodimers or bifunctional synthases integrated into broader terpenoid pathways.^[16]^[17]

Integration in Isoprenoid Pathways

Geranyl pyrophosphate (GPP) integrates into isoprenoid biosynthetic networks as a central C10 intermediate, derived from the condensation of isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP), the universal C5 building blocks. These precursors originate upstream from two distinct pathways: the mevalonate (MVA) pathway, which operates in the cytosol of eukaryotes and animals to generate IPP from acetyl-CoA through a series of seven enzymatic steps, followed by isomerization to DMAPP; and the methylerythritol phosphate (MEP) pathway, which functions in the plastids of plants and throughout bacteria, producing IPP and DMAPP directly from pyruvate and glyceraldehyde 3-phosphate in a balanced ratio.^[18]^[19] In bacteria, the MEP pathway serves as the exclusive source for these units, underscoring its essential role in prokaryotic isoprenoid metabolism.^[20] Organism-specific variations highlight the compartmentalized nature of these networks, particularly in plants, where dual pools of precursors exist: the cytosolic MVA pathway supports sesquiterpenoid and polyterpenoid synthesis, while the plastidial MEP pathway fuels monoterpenoids and carotenoids, with GPP bridging these compartments through limited metabolite exchange.^[19] Downstream, GPP extends the chain as a substrate for farnesyl pyrophosphate synthase (FPPS), which catalyzes its condensation with an additional IPP molecule to yield farnesyl pyrophosphate (FPP), a C15 precursor for longer isoprenoids.^[21] This sequential elongation exemplifies the modular architecture of isoprenoid pathways, enabling diverse product formation. Regulation of GPP integration occurs through feedback mechanisms, where downstream isoprenoids such as GPP and FPP inhibit upstream enzymes like mevalonate kinase, preventing precursor overaccumulation and maintaining flux balance.^[22] In microbial genomes, particularly those of Streptomyces species, isoprenoid biosynthetic genes are frequently clustered, promoting coordinated regulation and evolution of specialized pathways.^[23] Evolutionarily, the prenyltransferases responsible for GPP formation and extension exhibit a conserved structural fold, including characteristic DDXXD motifs for metal ion coordination, across bacteria, plants, and animals, reflecting ancient origins and functional universality in isoprenoid assembly.^[24]

Biological Significance

Role as Terpenoid Precursor

Geranyl pyrophosphate (GPP) serves as the primary C₁₀ prenyl intermediate in the terpenoid biosynthetic pathway, acting as an essential precursor for the formation of monoterpenoids and contributing to the elongation of longer-chain isoprenoids.^[25] Synthesized by geranyl diphosphate synthase (GPPS) through the head-to-tail condensation of dimethylallyl pyrophosphate (DMAPP) and isopentenyl pyrophosphate (IPP), GPP functions as a prenyl donor by transferring its geranyl moiety to acceptor molecules, including additional IPP units to generate farnesyl pyrophosphate (FPP) and subsequently geranylgeranyl pyrophosphate (GGPP).^[25] This chain elongation process is crucial for producing higher terpenoids, such as carotenoids derived from GGPP in plastids of plants like red pepper, where GPP flux supports pigment biosynthesis.^[26] Although GPP can participate in the geranylation of certain biomolecules, such as tRNA in prokaryotes, in eukaryotes GPP's role is primarily in terpenoid assembly, while protein prenylation is mediated by longer-chain prenyl pyrophosphates such as FPP and GGPP.^[27] In monoterpene synthesis, GPP undergoes ionization and cyclization catalyzed by monoterpene synthases (TPS), yielding a diverse array of volatile compounds that serve ecological functions in plants. These enzymes initiate the reaction by cleaving the pyrophosphate group from GPP, forming a geranyl carbocation that rearranges into cyclic or acyclic structures, such as limonene, α-pinene, and menthol.^[28] For instance, in peppermint (Mentha piperita) trichomes, GPP-derived menthol contributes to essential oil production, while in conifers like Abies grandis, it leads to pinene formation.^[29] Similarly, limonene biosynthesis in citrus plants relies on GPP as the direct substrate for TPS activity, highlighting its role in generating aromatic volatiles that deter herbivores and attract pollinators.^[28] Metabolic engineering efforts underscore the importance of GPP flux in terpenoid production, with optimizations increasing precursor availability to boost yields. In engineered Escherichia coli expressing the mevalonate pathway and mint limonene synthase, GPP flux enhancements achieved limonene titers of 605 mg/L, demonstrating scalable monoterpene output.^[28] In yeast, fusing GPPS with TPS enzymes has similarly elevated pinene production by channeling IPP/DMAPP toward GPP, though linker lengths influence efficiency. Genetic perturbations further reveal GPP's impact; in Arabidopsis thaliana mutants lacking functional heteromeric GPPS subunits (e.g., GGPPS11 or GGPPS12), monoterpene emissions in flowers dropped by 40-70%, altering volatile profiles and reducing total monoterpene levels to as low as 631 peak area/mg fresh weight compared to wild-type controls.^[30] These deficiencies confirm GPPS as a bottleneck, with overexpression restoring or enhancing terpenoid diversity in model plants.^[30]

Antimicrobial Effects

Geranyl pyrophosphate (GPP) exhibits toxicity to bacterial cells, most notably Escherichia coli, where intracellular accumulation at moderate levels disrupts cellular function and inhibits growth. This toxicity manifests as reduced cell viability and metabolic imbalance, primarily through interference with the isoprenoid biosynthesis pathway. In engineered bacteria expressing the mevalonate pathway, GPP competitively binds to the ATP-binding site of mevalonate 5-kinase (M5K), inhibiting the enzyme and halting flux through the mevalonate pathway; native bacteria utilize the methylerythritol phosphate (MEP) pathway instead.^[31] The mechanism of GPP-induced growth arrest involves both enzymatic inhibition and potential membrane perturbation, as evidenced by studies in engineered E. coli strains for terpenoid production. Accumulation of GPP leads to feedback regulation that diverts resources from essential processes, resulting in stalled cell division and decreased biomass. In synthetic biology applications, such as monoterpene biosynthesis, GPP toxicity is a key bottleneck, with cellular uptake studies showing rapid intracellular buildup that correlates with diminished growth rates compared to control strains. Longer-chain analogs like farnesyl pyrophosphate (FPP) display similar but often more pronounced effects.^[31] Experimental evidence highlights toxicity thresholds for GPP-related compounds in the range observed during pathway engineering, where moderate doses (proxied by geraniol equivalents at ~0.05% v/v or 445 mg/L) significantly impair E. coli growth.^[32] In plants, downstream monoterpenoids derived from GPP contribute to antimicrobial defenses against pathogens. Bacterial resistance to prenyl pyrophosphate toxicity frequently involves efflux pumps, such as AcrAB-TolC, which export the compound and restore cellular homeostasis in tolerant strains. A seminal 2013 synthetic biology study on limonene production in E. coli identified key toxicity thresholds, demonstrating that unbalanced GPP levels lead to pathway inefficiency and growth arrest unless mitigated by optimized precursor supply.^[32]^[33] Recent engineering efforts as of 2025 include strategies to counter GPP toxicity, such as efflux modulation, to enable stable pinene production in E. coli.^[34]

Occurrence and Production

Natural Distribution

Geranyl pyrophosphate (GPP), a key C10 isoprenoid intermediate, is widely distributed across plant kingdoms, where it is synthesized notably in essential oil-producing species. In plants such as lavender (Lavandula angustifolia) and citrus (Citrus spp.), GPP serves as a primary precursor for monoterpene volatiles, with synthesis occurring in glandular trichomes and secretory structures of flowers and leaves. For instance, in lavender, GPP is predominantly synthesized and utilized in floral oil glands, where enzyme activity peaks during periods of high volatile emission, contributing to the plant's characteristic scent profile. These pools are primarily localized in plastids via the methylerythritol phosphate (MEP) pathway, though minor cytosolic contributions occur through crosstalk with the mevalonate pathway, enabling flux toward diverse terpenoids.^[35]^[36]^[37] In animals, GPP exists at low, transient levels as an obligatory intermediate in the mevalonate pathway, primarily supporting cholesterol and steroid biosynthesis. It is formed by the condensation of isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) via geranyl pyrophosphate synthase (GPPS), with expression regulated in specific tissues such as the midgut of insects like bark beetles (Ips pini), where it serves as the direct precursor for monoterpene pheromones. Overall abundance remains minimal due to rapid downstream conversion, preventing significant accumulation in mammalian or other vertebrate systems during sterol synthesis.^[16]^[38] Microbial production of GPP varies by domain and pathway, typically as a short-lived intermediate. In bacteria, GPP is generated transiently through the MEP pathway from pyruvate and glyceraldehyde-3-phosphate, serving as a building block for ubiquinone, carotenoids, and cell wall components without substantial buildup. Archaea, conversely, rely on a modified mevalonate pathway for GPP synthesis, using it to elongate into geranylgeranyl chains for ether lipid membranes, again with low steady-state levels due to efficient enzymatic turnover. Engineered microbial strains, such as Escherichia coli modified with overexpressed GPPS and geraniol synthase, increase flux through GPP to enhance terpenoid yields such as geraniol, far exceeding natural microbial concentrations.^[39]^[40] Detection of GPP in biological samples relies on advanced metabolomics techniques, including liquid chromatography-high-resolution mass spectrometry (LC-HRMS) coupled with isotope dilution for quantification. Stable isotope labeling, such as with uniformly ^{13}C-glucose, enables isotopologue profiling to track GPP flux and pool sizes, with limits of detection as low as 0.01 pmol and linear quantification up to 50 pmol. Sample quenching via fast filtration followed by extraction in isopropanol-water buffers preserves labile pyrophosphates, allowing precise measurement of plastidial or cytosolic fractions in plants and microbes.^[41] Ecologically, GPP underpins plant defense by fueling the biosynthesis of volatile terpenoids that deter herbivores and recruit beneficial predators. In response to herbivory, GPP-derived monoterpenes like linalool and limonene are emitted from damaged tissues, signaling neighboring plants to prime defenses or attracting parasitoids such as wasps that target pests on crops like rice. This indirect defense mechanism enhances ecosystem resilience, with GPP's role in the MEP pathway ensuring rapid volatile production during stress.^[42]

Synthetic Methods

Geranyl pyrophosphate (GPP) can be synthesized chemically from geraniol through a two-step process involving chlorination followed by phosphorylation. In the first step, geraniol is converted to geranyl chloride using N-chlorosuccinimide and dimethyl sulfide in dichloromethane at low temperature, yielding geranyl chloride in 93% efficiency.^[10] The geranyl chloride is then reacted with tris(tetrabutylammonium) hydrogen pyrophosphate in acetonitrile at room temperature to form the diphosphate, which is subsequently purified to the trisammonium salt.^[10] This method achieves an overall yield of approximately 80% from geraniol and avoids the low yields and side products associated with earlier approaches using pyrophosphoryl chloride directly on alcohols.^[10] Biocatalytic production of GPP relies on recombinant geranyl diphosphate synthase (GPPS), which catalyzes the head-to-tail condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). In vitro synthesis involves expressing GPPS genes, such as the small subunit from mint (Mentha spicata) or animal orthologs like the one from whitefly (Bemisia tabaci), in Escherichia coli, followed by purification and incubation with IPP and DMAPP substrates.^[16]^[14] These recombinant enzymes produce GPP with high specificity, often exceeding 90% conversion efficiency in optimized assays.^[16] For microbial fermentation, engineered hosts like E. coli or yeast are used, where GPPS is overexpressed alongside pathways supplying IPP and DMAPP, such as the mevalonate or methylerythritol phosphate routes, sometimes supplemented with external feeds of the precursors.^[43] This approach has enabled GPP accumulation in cell lysates, though direct isolation remains challenging due to hydrolysis. Recent advances as of 2023 include optimized fusion enzymes and pathway balancing to improve monoterpenoid precursor flux in microbial cell factories.^[43]^[40] Scaling up GPP production faces key challenges, including its chemical instability, as the pyrophosphate linkage hydrolyzes readily in aqueous conditions, leading to geraniol formation. Chemical synthesis scales well in laboratory settings with multi-gram yields, but industrial adaptation requires inert atmospheres and low temperatures to maintain integrity.^[10] Biocatalytic methods in microbial systems improve productivity through controlled IPP/DMAPP supplementation.^[43] Purification typically employs ion-exchange chromatography on resins like Dowex AG 50W-X8, followed by precipitation with organic solvents such as acetonitrile/isopropyl alcohol mixtures, achieving recoveries of 80–90%.^[10] Historical methods for GPP preparation date back to the mid-20th century, relying on extractions from plant tissues or crude chemical condensations, which often yielded impure mixtures contaminated by hydrolysis products. Post-2000 advances in biotechnology, including recombinant GPPS expression and pathway engineering, have shifted toward sustainable, enzyme-driven production, surpassing early chemical routes in specificity and environmental compatibility.^[16]^[43] Purity standards for GPP vary by application: research-grade material, often as lithium or ammonium salts, requires ≥95% purity by TLC or HPLC to ensure accurate enzymatic assays, with impurities like unreacted precursors below 1%. Commercial preparations for biotechnology uses demand similar high purity to avoid inhibiting downstream terpenoid synthases, typically verified by NMR and mass spectrometry.^[44]

Precursor Molecules

Isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) serve as the immediate C5 precursors to geranyl pyrophosphate in isoprenoid biosynthesis, functioning as the fundamental building blocks for all terpenoids. IPP, with its structure featuring a terminal double bond and a pyrophosphate group, acts as the nucleophilic donor unit that extends the prenyl chain. DMAPP, an isomer of IPP, provides the electrophilic allylic starter unit essential for initiating condensation reactions. These molecules are present in virtually all organisms and are interconvertible, with DMAPP primarily formed through the reversible isomerization of IPP catalyzed by isopentenyl pyrophosphate isomerase (IDI).^[45]^[46] The structural relationship between IPP and DMAPP enables their head-to-tail linkage, where the C1 position of DMAPP (the "head") bonds to the C4 position of IPP (the "tail"), releasing pyrophosphate and forming a new trans double bond. This mechanism inherits the stereochemistry of the precursor double bonds, maintaining the characteristic E-configuration in the resulting linear chain and ensuring geometric uniformity in downstream isoprenoids. The allylic nature of DMAPP facilitates this SN1-like displacement, with the positive charge delocalized across its double bond system during the reaction.^[47]^[48] IPP and DMAPP originate from two independent biosynthetic routes conserved across taxa: the mevalonate pathway in eukaryotes and some bacteria, which converts three units of acetyl-CoA to IPP via mevalonic acid intermediates; and the methylerythritol phosphate (MEP) pathway in plastids, archaea, and most bacteria, starting from glyceraldehyde-3-phosphate and pyruvate to yield 1-deoxy-D-xylulose 5-phosphate as the first committed intermediate. These pathways provide the necessary pool of precursors, with IDI ensuring balanced IPP/DMAPP ratios for efficient chain elongation. Rare structural analogs, such as lavandulyl pyrophosphate, arise from atypical condensations of IPP and DMAPP, producing branched C10 variants observed in select monoterpenoids like lavandulol.^[49]^[50]^[51]

Derivative Isoprenoids

Geranyl pyrophosphate (GPP) serves as a central intermediate in the biosynthesis of longer-chain isoprenoids through enzymatic chain elongation. Farnesyl pyrophosphate (FPP, C₁₅), a key sesquiterpene precursor, is formed by the action of farnesyl pyrophosphate synthase (FPPS), which catalyzes the condensation of GPP with an additional molecule of isopentenyl pyrophosphate (IPP). This reaction extends the prenyl chain by one isoprene unit, producing FPP as a substrate for downstream sesquiterpene synthesis and other metabolic processes.^[21] FPPS exhibits a sequential mechanism where the allylic substrate (GPP) binds first, followed by IPP, leading to the release of pyrophosphate and formation of the new C-C bond.^[52] Further extension of the chain yields geranylgeranyl pyrophosphate (GGPP, C₂₀), primarily through the activity of geranylgeranyl pyrophosphate synthase (GGPPS), which adds another IPP unit to FPP. GGPP acts as the foundational precursor for diterpenes, including gibberellins in plants and retinal in animals, as well as carotenoids and other polyprenylated compounds.^[53] In some organisms, such as plants and bacteria, GGPPS can directly utilize shorter chains like GPP under specific conditions, but the predominant pathway involves stepwise elongation via FPP. This C₂₀ intermediate is crucial for the structural diversity of diterpenoids, which exhibit roles in growth regulation and defense.^[2] Beyond terpenoid backbones, GPP-derived compounds participate in protein post-translational modifications, particularly prenylation, which anchors proteins to membranes for signaling functions. Although GPP can theoretically enable geranylation (C₁₀ attachment), this is rare in eukaryotic systems; instead, prenylation predominantly involves farnesylation with FPP or geranylgeranylation with GGPP. For example, G-protein γ-subunits and Rho family GTPases such as RhoA undergo geranylgeranylation using GGPP, facilitating their localization to the plasma membrane and activation in cytoskeletal regulation, cell migration, and vesicular trafficking.^[54] Disruption of this geranylgeranylation impairs Rho-mediated signaling pathways, highlighting its specificity.^[55] GPP also contributes to monoterpene formation (C₁₀) through dephosphorylation or cyclization. For instance, hydrolysis of GPP by geraniol synthase yields geraniol, a volatile alcohol with antimicrobial properties found in essential oils of plants like lemongrass and geranium. In contrast, FPP-derived sesquiterpenes (C₁₅), such as farnesol or cyclic forms like humulene, arise from ionization and cyclization reactions catalyzed by sesquiterpene synthases, serving roles in plant defense and aroma compounds.^[56] The chain length influences biological activity; shorter C₁₀ monoterpenes like geraniol exhibit higher volatility and moderate toxicity to microbes, whereas longer C₁₅ sesquiterpenes from FPP display enhanced membrane permeability and potent anti-inflammatory effects, with geranylgeranylated derivatives showing greater cytotoxicity in cancer cells due to altered prenylation specificity.^[57]

Research and Applications

Historical Discovery

Geranyl pyrophosphate (GPP), an essential intermediate in isoprenoid biosynthesis, was first identified in the late 1950s during investigations into the mevalonate pathway for cholesterol and terpenoid synthesis. Pioneering work by Feodor Lynen and Konrad Bloch, who shared the 1964 Nobel Prize in Physiology or Medicine for their discoveries on cholesterol metabolism, revealed GPP as the product of head-to-tail condensation between dimethylallyl pyrophosphate and isopentenyl pyrophosphate. In 1958–1959, Lynen's laboratory isolated and characterized GPP enzymatically from yeast extracts, confirming its role as a C10 precursor to longer-chain isoprenoids like farnesyl pyrophosphate.^[58] This breakthrough built on earlier 1950s studies by Bloch on acetate incorporation into cholesterol and Lynen's elucidation of acetyl-CoA as the pathway's starting unit. Key milestones in the 1960s included the development of enzymatic assays that validated GPPS activity, the enzyme catalyzing GPP formation. In 1966, researchers purified and characterized GPPS from pig liver, demonstrating its specificity for producing the trans isomer of GPP and distinguishing it from downstream farnesyl pyrophosphate synthase.^[59] Complementary stereochemical studies by John Cornforth and George Popják during this decade clarified the reaction mechanism, showing inversion at the allylic carbon during condensation, which informed broader understanding of terpenoid assembly. These efforts established GPP's foundational position in the mevalonate-dependent route, with seminal papers such as Lynen's 1959 review in Angewandte Chemie outlining the pathway's intermediates.^[58] The 1980s marked advances in molecular biology, including the cloning of prenyl synthase genes that extended knowledge of GPP-related enzymes; for example, the yeast farnesyl pyrophosphate synthase gene was cloned in 1989, enabling heterologous expression and functional studies.^[60] Nomenclature for the compound evolved from "geranyl pyrophosphate" in early biochemical literature to "geranyl diphosphate" (GDP) in modern contexts, reflecting its diphosphate ester structure and alignment with isopentenyl diphosphate conventions. However, early research overlooked an alternative non-mevalonate pathway for isoprenoid precursors, the 2-C-methyl-D-erythritol 4-phosphate (MEP) route, which was not discovered until the 1990s through labeling experiments by Michel Rohmer and colleagues in bacteria and plants.^[61]

Modern Uses in Biotechnology

In metabolic engineering, geranyl pyrophosphate (GPP) serves as a critical precursor for monoterpene biosynthesis, with overexpression or modification of geranyl pyrophosphate synthase (GPPS) in microbial hosts enabling enhanced production of valuable compounds. In Saccharomyces cerevisiae, engineering the endogenous farnesyl pyrophosphate synthase (Erg20p) into a GPPS variant has significantly increased monoterpene titers, such as geraniol and limonene, by redirecting flux from farnesyl pyrophosphate toward the C10 pathway, achieving up to 3.4-fold improvements in yields for potential use in biofuels and flavors.^[62]^[63] Similar strategies in Escherichia coli involve bioprospecting GPPS enzymes and optimizing flux through the methylerythritol phosphate pathway, resulting in gram-per-liter production of geraniol, a key fragrance ingredient, while addressing precursor imbalances.^[64] These approaches prioritize compartmentalization and dynamic regulation to minimize competition with native pathways, supporting scalable production for industrial applications. In synthetic biology, 2010s research highlighted GPP's toxicity in microbial hosts, where accumulation disrupts membrane integrity and growth, prompting strategies like dominant-negative GPPS mutants to control flux and prevent buildup during monoterpene synthesis.^[65] Flux optimization in E. coli using machine learning-guided translational control of GPPS and downstream synthases has enabled predictive chassis design, boosting geraniol titers by over 100-fold while mitigating toxicity through esterification or export mechanisms.^[66] These tools have facilitated orthogonal pathways in yeast, decoupling monoterpene production from endogenous metabolism for applications in microbial control and high-value chemical synthesis.^[67] Industrial examples include the use of engineered yeast strains overexpressing GPPS to produce artemisinin precursors, where balanced IPP/DMAPP pools enhance amorphadiene yields from downstream farnesyl pyrophosphate, contributing to semi-synthetic antimalarial production.^[68] For fragrances, GPPS engineering in S. cerevisiae and E. coli has scaled geraniol output to levels suitable for commercial flavor and perfume industries, with titers exceeding 1 g/L in optimized fermentations.^[69] Pharmaceutically, GPP's role in the prenylation pathway positions upstream inhibitors, such as statins targeting HMG-CoA reductase, as candidates for cancer therapy by depleting isoprenoid pools and blocking protein farnesylation/geranylgeranylation essential for tumor signaling.^[70] Similarly, bisphosphonate inhibitors of related synthases like farnesyl pyrophosphate synthase exhibit antimicrobial effects by disrupting terpenoid biosynthesis in pathogens, with potential extensions to GPPS modulation for controlling bacterial prenylation-dependent virulence.^[71] Future prospects involve CRISPR-based edits for precise pathway tuning, such as multiplexed activation of GPPS in plants to redirect flux toward sustainable monoterpene synthesis, alongside 2020s advances in yeast platforms achieving universal terpenoid production through modular synthetic operons.^[72]^[73] As of 2025, advances in microbial production of geraniol include engineered Escherichia coli strains achieving 13.2 g/L and Saccharomyces cerevisiae 5.5 g/L, supporting industrial scale-up for sustainable terpenoid synthesis.^[74] These developments promise eco-friendly bioproduction, reducing reliance on chemical extraction for pharmaceuticals and biofuels.

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Insufficient relevant content. The provided text only includes a PubChem identifier (CID 445995), molecular formula (C10H20O7P2), and a reference to Geranyl Diphosphate, along with a JavaScript requirement notice and a logo. No detailed information on molecular weight, IUPAC name, other names, or structure description (carbon chain, double bonds, configuration, pyrophosphate group) is present.
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trisammonium geranyl diphosphate - Organic Syntheses Procedure
The IR, 1H NMR, and 13C NMR spectra of this material are identical to those for distilled geranyl chloride (bp 49–51°C at 0.2 mm). Distillation on a small scale ...
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Synthesis of geranyl pyrophosphate (GPP) in plants - ResearchGate
The present review highlights the Foeniculum vulgare essential oil extraction process, chemical composition, and various applications with the biological ...
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Geranyl diphosphate synthase: Cloning, expression, and ... - PNAS
Geranyl diphosphate synthase, which catalyzes the condensation of dimethylallyl diphosphate and isopentenyl diphosphate to geranyl diphosphate,Missing: review | Show results with:review
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Structure, mechanism and function of prenyltransferases - FEBS Press
Jul 4, 2002 · This enzyme catalyzes the transfer of the carbon moiety of FPP or GGPP to a conserved cysteine residue in a CaaX motif of protein and peptide ...<|control11|><|separator|>
[14]
Structure of a Heterotetrameric Geranyl Pyrophosphate Synthase ...
This work presents the crystal structure of mint heteromeric prenyltransferase, which is responsible for the biosynthesis of geranyl pyrophosphate.
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A cytosolic bifunctional geranyl/farnesyl diphosphate synthase ... - NIH
May 1, 2023 · A cytosolic bifunctional geranyl/farnesyl diphosphate synthase provides MVA-derived GPP for geraniol biosynthesis in rose flowers.
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Isolation and functional expression of an animal geranyl ... - PNAS
Geranyl diphosphate synthase (GPPS) catalyzes the condensation of dimethylallyl diphosphate and isopentenyl diphosphate to form geranyl diphosphate.
[17]
A Bifunctional Geranyl and Geranylgeranyl Diphosphate Synthase Is ...
A bifunctional geranyl and geranylgeranyl diphosphate synthase is involved in terpene oleoresin formation in picea abies.
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Two-step pathway for isoprenoid synthesis - PMC - PubMed Central
Dec 24, 2018 · The mevalonate (MVA) pathway involves seven reactions to produce IPP from acetyl-CoA. DMAPP is then produced through the isomerization of IPP by ...
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Distinct Light-Mediated Pathways Regulate the Biosynthesis and ...
The plastidial MEP pathway produces IPP and DMAPP for the biosynthesis of photosynthesis-related isoprenoids (carotenoids and the side chains of chlorophylls, ...Missing: pyrophosphate | Show results with:pyrophosphate
[20]
The MEP pathway: Promising drug targets in the fight against ...
The MEP pathway consists of seven enzymatic steps starting from pyruvate (Pyr) and Dglyceraldehyde-3-phosphate (D-GAP) to produce IPP and DMAPP (Figure 2).
[21]
Farnesyl pyrophosphate synthase: a key enzyme in isoprenoid ...
Jan 25, 2013 · A key enzyme in this pathway is farnesyl pyrophosphate (EC 2.5.1.10) synthase (FPPS) which supplies precursors for the biosynthesis of essential isoprenoids.Missing: geranyl | Show results with:geranyl
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Negative Feedbacks by Isoprenoids on a Mevalonate Kinase Expressed in the Corpora Allata of Mosquitoes - PubMed
### Summary of Feedback Inhibition in Isoprenoid Pathway Involving GPP
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Genomic insights into the evolution of hybrid isoprenoid biosynthetic ...
Nov 17, 2015 · One hundred and twenty Streptomyces genomes were searched for HI biosynthetic gene clusters using ABBA prenyltransferases (PTases) as queries.
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Structure, catalysis, and inhibition mechanism of prenyltransferase
Productions of isoprenoids are catalyzed by a group of enzymes known as prenyltransferases, such as farnesyltransferases, geranylgeranyltransferases, terpenoid ...
[25]
The functional evolution of architecturally different plant geranyl ...
In plants, GPP is predominantly produced in plastids for the biosynthesis of monoterpenes, which are major constituents of plant volatiles, while FPP is ...
[26]
https://doi.org/10.1021/acs.jafc.8b04052
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Discovery and biological characterization of geranylated RNA ... - NIH
coli, and purified SelU protein in the presence of geranyl pyrophosphate and tRNA can produce geranylated tRNA. The presence or absence of the geranyl group ...
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https://doi.org/10.1105/tpc.109.071738
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https://doi.org/10.1016/j.molp.2015.05.001
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https://doi.org/10.1039/d1np00025j
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https://doi.org/10.1021/sb4001382
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https://doi.org/10.1016/j.ymben.2013.05.004
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https://pmc.ncbi.nlm.nih.gov/articles/PMC11747701/
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Natural Compounds in the Battle against Microorganisms—Linalool
Oct 15, 2022 · In plants, linalool is made from geranyl pyrophosphate (GPP)—a ... Lavender essential oil and its main constituents inhibit the ...
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https://doi.org/10.3390/cimb47070517
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https://doi.org/10.3390/molecules27206928
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https://doi.org/10.1016/j.isci.2018.10.019
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Biosynthesis of archaeal membrane ether lipids - Frontiers
In general, IPP and DMAPP are synthesized by the MEP pathway in most of the bacteria and by two MVA pathways in eukarya and archaea. The classical MVA pathway ...
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https://doi.org/10.1016/j.ymben.2021.04.008
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Functional analysis of isoprenoid precursors biosynthesis by ... - NIH
Aug 21, 2019 · IPP is isomerized into dimethylallyl diphosphate (DMAPP), which is then condensed with IPP to generate geranyl pyrophosphate (GPP). Longer ...
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https://doi.org/10.3390/ijms241411561
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US8715962B2 - Production of geranyl diphosphate - Google Patents
In one biosynthetic pathway, GPP is produced from the five-carbon precursors isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP).
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https://www.sigmaaldrich.com/US/en/product/sial/19533
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The Non-mevalonate Pathway of Isoprenoid Precursor Biosynthesis
Isopentenyl pyrophosphate (IPP) and dimethylallyl pyrophosphate (DMAPP) are the universal precursors of natural products called isoprenoids. This large family, ...
[46]
Differential expression of two isopentenyl pyrophosphate ... - PNAS
The enzyme isopentenyl pyrophosphate (IPP) isomerase catalyzes the reversible isomerization of IPP to produce dimethylallyl pyrophosphate, the initial ...
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Isoprenoids in three-dimensional space: the stereochemistry of ...
Oct 7, 2011 · This review summarises the accumulated knowledge about the stereochemical courses of biosynthetic reactions to isoprenoids . Important pathways ...Missing: inheritance head- tail linkage
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Towards efficient terpenoid biosynthesis: manipulating IPP and ...
Feb 6, 2019 · We first introduce biosynthetic pathways of IPP and DMAPP, and then summarize the current strategies adopted for manipulating IPP and DMAPP supply.Missing: delta | Show results with:delta<|separator|>
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ENZYMES OF THE MEVALONATE PATHWAY OF ISOPRENOID ...
The mevalonate pathway accounts for conversion of acetyl-CoA to isopentenyl 5-diphosphate, the versatile precursor of polyisoprenoid metabolites and natural ...
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The Multifaceted MEP Pathway: Towards New Therapeutic ... - NIH
Moreover, the impact of the double bond configuration of the isoprene unit has been studied, and the PAgs in their natural E configuration turned out to be ...
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Synthetic Biology, Combinatorial Biosynthesis, and Chemo ...
For example, there are four possible C10 linear monoterpenes produced from DMAPP/IPP: GPP, NPP (the cis conformer of GPP), lavandulyl pyrophosphate, made from a ...
[52]
Investigation of the catalytic mechanism of farnesyl pyrophosphate ...
Farnesyl pyrophosphate synthase (FPPS) catalyses the formation of a key cellular intermediate in isoprenoid metabolic pathways, farnesyl pyrophosphate, ...
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Geranylgeranyl diphosphate synthase: Role in human health ...
Jan 17, 2023 · Geranylgeranyl diphosphate synthase (GGDPS), an enzyme in the isoprenoid biosynthesis pathway, is responsible for the production of geranylgeranyl ...
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Inhibition of protein geranylgeranylation and RhoA/RhoA kinase ...
Geranylgeranylation of RhoA small G-protein is essential for its localization to cell membranes and for its biological functions. Many RhoA effects are ...Missing: geranylation geranyl
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De novo production of six key grape aroma monoterpenes by ... - NIH
This metabolic manipulation was possible because plant MTPSs catalyze the synthesis of monoterpenes from geranyl pyrophosphate (GPP) in a single step, and S.
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A Novel Pathway for Sesquiterpene Biosynthesis from Z,Z-Farnesyl ...
It has been found to date that the biosynthesis of terpenoids proceeds from IPrPs in the E configuration via the action of a conserved family of enzymes, the ...Missing: geranyl | Show results with:geranyl
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[PDF] Konrad Bloch - The biological synthesis of cholesterol - Nobel Prize
The discovery of geranylpyrophosphate53 and of farnesylpyro- phosphate50 in Lynen's laboratory completed the description of the biosyn- thetic pathway and of ...Missing: geranyl | Show results with:geranyl
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The purification and properties of pig liver geranyl pyrophosphate ...
The purification and properties of pig liver geranyl pyrophosphate synthetase. J Biol Chem. 1966 Nov 25;241(22):5353-60. Authors. J K Dorsey, J A Dorsey, J W ...
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The Purification and Properties of Pig Liver Geranyl Pyrophosphate ...
The purification and properties of geranyl pyrophosphate synthetase are reported. This enzyme is purified simultaneously with the enzyme for the synthesis ...<|control11|><|separator|>
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The discovery of a mevalonate-independent pathway for isoprenoid ...
The discovery of a mevalonate-independent pathway for isoprenoid biosynthesis in bacteria, algae and higher plants. Nat Prod Rep. 1999 Oct;16(5):565-74.Missing: MEP 1990s
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Engineering Monoterpene Production in Yeast Using a Synthetic ...
We engineered Erg20p into a geranyl diphosphate synthase and achieved a significant increase in monoterpene titers.
[62]
Orthogonal monoterpenoid biosynthesis in yeast constructed on an ...
Aug 23, 2019 · Therefore, Erg20p is very efficient in utilizing GPP, reducing the GPP pool and hindering monoterpene production. Thus, we set out to engineer ...
[63]
Engineering Escherichia coli for production of geraniol by systematic ...
Since toxic IPP is the precursor for GPP, growth inhibition could be effectively alleviated by GPPS overexpression. On the other hand, the upregulation of GPPS ...
[64]
Synthetic and systems biology for microbial production of commodity ...
Apr 7, 2016 · The toxicity of these compounds was evaluated using strains that accumulate isopentenyl pyrophosphate, GPP, and FPP, and the accumulation of FPP ...
[65]
Machine Learning of Designed Translational Control Allows ...
Dec 18, 2018 · This methodology is potentially compatible with any biochemical pathway and provides a powerful tool toward predictive design of bacterial production chassis.
[66]
Engineering Saccharomyces cerevisiae for the production of the ...
Jun 5, 2018 · Some reports also demonstrated the potential of metabolic engineering for monoterpene production in S. cerevisiae [18]. GPP is synthesized by ...
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A Novel Semi-biosynthetic Route for Artemisinin Production Using ...
Mar 9, 2009 · Because isopentenyl and dimethylallyl pyrophosphates are the universal precursors to all isoprenoids, the strains developed in this study can ...
[68]
Efficient production of (S)-limonene and geraniol in Saccharomyces ...
We generated a novel Erg20 WWG mutant with over four times higher GPP accumulating capability than Erg20 WW, as observed through geraniol production levels.<|control11|><|separator|>
[69]
Isoprenoids responsible for protein prenylation modulate the ...
Dec 20, 2017 · Our data indicate that the anticancer effects of statins observed in numerous studies to a large extent are mediated through isoprenoid intermediates of the ...
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Characterization of Potential Drug Targets Farnesyl Diphosphate ...
We report here that Schistosoma mansoni farnesyl diphosphate synthase (SmFPPS) and geranylgeranyl diphosphate synthase (SmGGPPS) are potential drug targets for ...
[71]
Engineering a universal and efficient platform for terpenoid ... - PNAS
Dec 28, 2022 · Sequential addition of IPP units to DMAPP generates terpenoid precursors, like geranyl diphosphate (GPP, C10), farnesyl diphosphate (FPP, C15), ...
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[PDF] Application and development of CRISPR technology in ... - Frontiers
Jan 9, 2025 · This article introduces the principles of CRISPR technology and its efficacy in gene editing, followed by a detailed discussion of its ...Missing: geranyl | Show results with:geranyl<|control11|><|separator|>