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Ginsenoside

Ginsenosides are a diverse class of , primarily of the dammarane type, that serve as the main pharmacologically active compounds in plants of the genus , such as (Asian ginseng) and (American ). These glycosides, first isolated in 1963, consist of a triterpenoid aglycone backbone attached to one or more moieties, with over 270 distinct structures identified as of 2024, commonly classified into protopanaxadiol (e.g., Rb1, Rb2, Rg3) and protopanaxatriol (e.g., Rg1, ) groups based on their aglycone types. Found predominantly in the roots but also in other plant parts like leaves and fruits, ginsenoside content varies by species, plant age (peaking around 6 years), cultivation method, harvest season, and processing techniques, such as steaming to produce red ginseng, which enhances certain bioactive forms. Ginsenosides have been central to the traditional use of in for over 2,000 years, where the herb is valued for its adaptogenic properties to promote and to . Pharmacologically, they exhibit a broad spectrum of effects through multi-target mechanisms, including modulation of receptors (e.g., and receptors), enhancement of defenses, and regulation of signaling pathways like PI3K/Akt and . Notable activities encompass against and neurodegeneration, effects by inhibiting pro-inflammatory cytokines, cardiovascular benefits such as improved endothelial function and reduced arrhythmias, antidiabetic actions via glucose regulation, and anticancer potential through induction of and inhibition of tumor . Their therapeutic promise has spurred extensive research, with ginsenosides increasingly explored as dietary supplements for immune modulation, metabolic health, and anti-aging, though challenges—due to poor and rapid metabolism—necessitate formulations like nanoparticles or hydrolysis for enhanced efficacy. Ongoing studies emphasize their role in veterinary applications for animal health and in for microbial production to meet demand, underscoring ginsenosides' significance in both traditional and modern .

Nomenclature and Definition

Etymology and Naming

The term "ginsenoside" is derived from "," referring to of the genus , combined with "" to denote their classification as triterpenoid glycosides that produce soap-like foaming when agitated in water, a characteristic property of . This nomenclature reflects their origin as bioactive compounds primarily isolated from ginseng roots, where they constitute the major secondary metabolites responsible for the plant's pharmacological effects. An alternative designation, "panaxosides," stems from the genus name , coined by Swedish botanist in the from the Greek word panax (Πάναξ), meaning "all-healing" or "," based on the herb's longstanding use in as a versatile tonic. The isolation and initial characterization of these compounds as ginsenosides occurred in 1963, with systematic structural studies advancing in the through research by Japanese scientists led by Sankichi Shibata and Osamu , who separated and identified key from . The conventional naming system for individual ginsenosides uses the prefix "Rg" or similar (with "R" denoting "root"), followed by lowercase letters (such as a, b, c) to group compounds by during chromatographic separation, and to indicate the order of or elution in . Over 150 distinct ginsenosides have been identified to date, as of , with Rb1 serving as a representative example of the most abundant protopanaxadiol-type compound in roots. As structural elucidations progressed, this empirical system evolved toward standardized International Union of Pure and Applied Chemistry (IUPAC) ; for instance, ginsenoside Rb1 is formally named 20-[β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyloxy]-12β-hydroxydammar-24-en-3β-yl β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside.

Chemical Definition

Ginsenosides are a class of steroid glycosides and triterpene found almost exclusively in species of the genus, such as and Panax quinquefolius. These compounds are characterized by their amphiphilic nature, arising from a hydrophobic triterpenoid aglycone core combined with hydrophilic sugar chains attached via glycosidic bonds. This structural duality contributes to their role as surface-active agents in plant tissues. The general chemical formula of ginsenosides involves a dammarane-type triterpenoid aglycone—a tetracyclic structure derived from dammarenediol—as the backbone, covalently linked to one or more units (such as glucose or ) through ether-type glycosidic linkages. Molecular weights of these glycosides typically range from 800 to 1200 , depending on the number and type of sugar moieties. Key physical properties of ginsenosides include a characteristic bitter taste, which serves as an in plants, and the ability to produce stable foams when agitated in water, a hallmark of their classification. They exhibit poor in water, though enhances their hydrophilicity compared to the non-glycosylated aglycones, and they are more readily soluble in polar organic solvents like . For analytical detection, ginsenosides show strong UV at approximately 203 nm due to the π-π* transitions in their aglycone chromophores. Ginsenosides are distinguished from other saponins by their exclusive occurrence in ginseng species and their predominant dammarane-based triterpenoid skeletons, in contrast to the steroidal (C27) frameworks of saponins from plants like those in the Liliaceae family.

Structural Classification

Aglycone Types

Ginsenosides are primarily classified into three main aglycone types based on their core triterpene skeletons: dammarane (the most common, comprising over 90% of known ginsenosides), oleanane, and ocotillol. The dammarane type dominates in species like Panax ginseng and Panax notoginseng, while oleanane and ocotillol types are less prevalent overall. These aglycones serve as the hydrophobic cores that are subsequently modified by glycosylation to yield the full ginsenoside structures. The dammarane-type aglycones, derived from the cyclization of 2,3-oxidosqualene, feature a tetracyclic structure with 17 carbon atoms in the ring system and a side chain at C-17. They are further subdivided into protopanaxadiol (PPD) and protopanaxatriol (PPT) subtypes. PPD aglycones, such as that in ginsenoside Rb1, possess hydroxyl groups at C-3, C-12, and C-20, with the molecular formula \ce{C30H52O3}. PPT aglycones, exemplified by that in ginsenoside Rg1, include an additional hydroxyl at C-6, resulting in the formula \ce{C30H52O4}. Stereochemistry at C-20 is typically 20S in naturally occurring forms, though 20R epimers can arise as artifacts during extraction or processing. Oleanane-type aglycones exhibit a pentacyclic structure akin to , characterized by a between C-12 and C-13 and a group at C-28. A representative example is the aglycone of ginsenoside Ro, which maintains this configuration and is found in lower abundance compared to dammarane types. Ocotillol-type aglycones are rare, constituting less than 1% of ginsenosides in , and consist of a tetraoxygenated dammarane featuring a tetrahydrofuran ring at C-20. An example is the aglycone in majonoside-R2, which includes oxygenations at C-20, C-24, and other positions, distinguishing it from standard dammarane subtypes.

Glycosylation Patterns

Ginsenosides are characterized by their at specific positions on the aglycone backbone, primarily through β-glycosidic bonds that link moieties, resulting in mono-, di-, or trisaccharide chains. These attachments occur mainly at C-3 for protopanaxadiol (PPD) and protopanaxatriol () types as well as oleanane types, at C-6 for types, and at C-20 for all dammarane-based ginsenosides (PPD and ). The common sugars involved include β-D-glucose (Glc), α-L-rhamnose (Rha), α-L-arabinose (Ara in pyranosyl or furanosyl forms), β-D-xylose (Xyl), and β-D-glucuronic acid (GlcA), which are attached via specific linkages such as (1→2), (1→6), or (1→4). In PPD-type ginsenosides, glycosylation typically features glucose chains at C-3 and C-20, contributing to their within the Rb, Rc, and Rd groups. For instance, ginsenoside Rb1 exhibits a at C-3 [β-D-Glc-(1→2)-β-D-Glc] and a at C-20 [β-D-Glc], enhancing its structural diversity within this subclass. PPT-type ginsenosides extend this pattern by incorporating an additional sugar at C-6, often a glucose-rhamnose combination, as seen in ginsenoside Re with β-D-Glc-(1→2)-α-L-Rha at C-6 and β-D-Glc at C-20, distinguishing the Rg and Re groups. Oleanane-type ginsenosides, such as Ro, are primarily glycosylated at C-3 with a glucuronic acid-arabinose [β-D-GlcA-(1→2)-β-D-Ara(p)], lacking the C-20 modification common in dammarane types. These glycosylation variations generate over 270 known ginsenoside variants through combinatorial attachments as of 2024, with the number and complexity of sugar chains directly influencing —more extensive , as in the Rb group with multiple glucoses, increases hydrophilicity compared to less substituted forms like the Rg group. This gradient affects and separation in analytical contexts, underpinning the structural subclassification of ginsenosides. Rare modifications, such as on hydroxyl groups (e.g., in the Rs series like Rs1 with an at the C-20 glucose) or sulfation in certain species, further diversify patterns but occur infrequently.
Ginsenoside TypeExampleGlycosylation at Key SitesSugars and Linkages
PPDRb1C-3: ; C-20: β-D-Glc-(1→2)-β-D-Glc at C-3; β-D-Glc at C-20
PPTReC-6: ; C-20: β-D-Glc-(1→2)-α-L-Rha at C-6; β-D-Glc at C-20
OleananeRoC-3: β-D-GlcA-(1→2)-β-D-Ara(p) at C-3

Biosynthesis in Plants

Biosynthetic Pathway

Ginsenosides are primarily synthesized through the mevalonate (MVA) pathway in the of cells, where is converted to isopentenyl diphosphate () and dimethylallyl diphosphate (DMAPP), the universal isoprenoid precursors. These units condense sequentially to form geranyl diphosphate (GPP, C10), then farnesyl diphosphate (FPP, C15), which serves as the immediate precursor for triterpenoid biosynthesis. Squalene synthase catalyzes the head-to-head dimerization of two FPP molecules to produce (C30), a key linear intermediate: $2 \text{ FPP} \rightarrow \text{squalene} + 2 \text{ PPi}. Squalene is then epoxidized at the 2,3-position by squalene epoxidase to yield (3S)-2,3-oxidosqualene, the substrate for cyclization reactions that determine the aglycone skeletons of ginsenosides. In the dammarane branch, predominant in , dammarenediol-II synthase cyclizes 2,3-oxidosqualene to dammarenediol-II, marking the first committed step toward protopanaxadiol (PPD)- and protopanaxatriol ()-type ginsenosides. Dammarenediol-II undergoes sequential cytochrome P450-mediated s: first at C-12 to form PPD, followed by an additional at C-6 to yield . The oleanane branch diverges earlier, with β-amyrin synthase cyclizing 2,3-oxidosqualene to β-amyrin, which is further oxidized (e.g., at C-28) to , the precursor to oleanane-type ginsenosides like Ro. Post-cyclization modifications involve at various positions on the aglycones, primarily using UDP-activated sugars such as UDP-glucose and UDP-rhamnose, catalyzed by UDP-glycosyltransferases (UGTs). These attachments occur at sites like C-3, C-6, and C-20, generating the diverse array of ginsenosides and enhancing their solubility and bioactivity. Although the MVA pathway dominates, the methylerythritol (MEP) pathway in plastids contributes minor isoprenoid precursors, with transcript levels indicating comparable flux to MVA in roots but higher in leaves. Environmental factors, such as elicitors like , influence pathway by upregulating early MVA genes (e.g., ), thereby increasing ginsenoside accumulation in elicited tissues.

Key Enzymes and Regulatory Genes

The of ginsenosides in involves several key upstream enzymes in the mevalonate (MVA) pathway, which provide for triterpenoid formation. (HMGR) acts as the rate-limiting enzyme in the MVA pathway, catalyzing the conversion of to mevalonate and influencing the flux toward isoprenoid essential for ginsenoside production. synthase (SQS) then condenses two molecules into , a committed step in triterpenoid , while epoxidase (SQE) oxidizes to 2,3-oxidosqualene, the immediate precursor for cyclization reactions. These enzymes are highly expressed in ginseng roots and respond to elicitors like , enhancing ginsenoside accumulation. Downstream cyclization and oxidation steps are mediated by oxidosqualene cyclases and monooxygenases. Dammarenediol-II synthase (DS, also known as PgDDS) is a critical oxidosqualene cyclase that cyclizes 2,3-oxidosqualene to dammarenediol-II, the foundational aglycone for most ginsenosides, showing specificity for the dammarane skeleton in species. Protopanaxadiol synthase activity is primarily carried out by the enzyme CYP716A47 (also referred to as PPD synthase), which hydroxylates dammarenediol-II at the C-12 position to form protopanaxadiol. Further oxidations involve the CYP716A family; CYP716A53v2 catalyzes the C-6 of protopanaxadiol to produce protopanaxatriol, a key step in the PPT-type ginsenoside branch. These P450s exhibit tissue-specific expression and are upregulated under stress conditions to modulate ginsenoside profiles. Glycosylation, which adds sugar moieties to aglycones, is catalyzed by UDP-glycosyltransferases (UGTs). Members of the UGT71A family, such as PgUGT71A53 (also denoted as UGTPg1), specifically attach to the C-20 hydroxyl group of protopanaxadiol and protopanaxatriol, forming compounds like ginsenoside C-K and F1, respectively, and representing a pivotal early step. The UGT74 family contributes to further diversification by adding or residues; for example, certain UGT74 enzymes transfer to the C-20 glucose of protopanaxadiol-type ginsenosides, enhancing structural complexity and . These UGTs are abundant in the genome, with over 200 identified, and their activities are coordinated to produce the diverse patterns observed in natural extracts. Regulatory genes, including transcription factors and non-coding RNAs, fine-tune ginsenoside by modulating expression. WRKY transcription factors, such as PgWRKY2, positively regulate DS expression, thereby promoting dammarenediol-II formation and overall ginsenoside yields in response to elicitors like . MYB and bHLH factors also influence the pathway; for instance, certain MYB proteins activate upstream MVA pathway genes, while bHLH members coordinate stress-induced responses that boost CYP716A expression. MicroRNAs contribute to post-transcriptional control, with miR171 targeting genes like CYP716A17 to repress oxidation steps under specific conditions. Recent studies (as of 2025) highlight microRNAs targeting genes and jasmonate-mediated upregulation in cell cultures to boost ginsenoside levels. Recent advances include CRISPR/Cas9-mediated knockout of PgDDS, which reduced ginsenoside yields by approximately 50% in transgenic lines, confirming its essential role and enabling pathway . Ginsenoside-related genes are organized in loose clusters within the genome, which was initially sequenced in 2017 and refined through 2023 assemblies revealing over 3.5 Gb of sequence with high content. These clusters feature proximal CYP716A and UGT genes, often with elicitor-responsive promoters that respond to or abiotic stresses, facilitating coordinated regulation without tight physical linkage typical of fungal pathways. This genomic architecture supports adaptive ginsenoside production in perennial roots.

Chemical Reactivity and Metabolism

General Chemical Properties and Reactions

Ginsenosides are triterpenoid characterized by low aqueous , typically around 1 mg/mL for major protopanaxadiol-type compounds like Rb1, due to their hydrophobic aglycone cores, though attached moieties increase and enhance limited compared to the aglycones alone. They exhibit lipophilic tendencies from the dammarane , facilitating in organic solvents such as (9.8–10.2 mg/mL for Rb1) and . Melting points generally range from 194–203°C across common ginsenosides, such as 197°C for Rb1 and 201–203°C for Re, reflecting their crystalline powder form under standard conditions. Ginsenosides demonstrate notable instability under acidic conditions, where glycosidic bonds undergo , particularly at low , leading to deglycosylation and formation of less polar derivatives or aglycones like compound K from protopanaxadiol types. They remain relatively heat-stable up to 100°C but degrade via β-elimination reactions above 120°C, with processes converting polar ginsenosides such as Rb1 into less polar forms like Rg3. Exposure to light induces , with significant observed after 30 days, while (ROS) promote oxidation, targeting double bonds and hydroxyl groups to form epoxides and other oxidized products, as seen in the aqueous instability of Rg5 where ~95% decomposes in 10 days at 25°C due to dissolved oxygen. Key chemical reactions of ginsenosides include acid- or base-catalyzed of glycosidic linkages at C-3 and C-20 positions, yielding aglycones such as protopanaxadiol or protopanaxatriol. Oxidation occurs at hydroxyl groups and sites, often under , leading to structural modifications like formation. is a rare transformation during high-temperature processing, such as in sun ginseng production, where acetyl groups attach to hydroxyls, forming derivatives like 3β,12β-diacetyl ginsenosides. Analytical identification relies on the foaming test, where ginsenosides produce persistent foam in aqueous solutions due to their amphiphilic , confirming their class. The Liebermann-Burchard reaction detects the aglycone moiety, yielding a color change from pink to blue-green upon treatment with and , specific for unsaturated triterpenoids. Temperature-dependent reactions highlight elimination pathways at elevated temperatures, as detailed in a 2025 kinetic study showing Rb1 transformation to Rg5 and Rk1 via deglycosylation and dehydration during steaming or hydrothermal processing, with rates accelerating above 120°C to favor less-polar rare ginsenosides.

In Vivo Metabolism

Ginsenosides undergo extensive biotransformation in vivo, primarily mediated by the gut microbiota and hepatic enzymes. In the gastrointestinal tract, gut bacteria play a crucial role in the deglycosylation of protopanaxadiol (PPD)-type ginsenosides, such as Rb1, through sequential hydrolysis of sugar moieties. Bacteroides species, among others, produce β-glucosidases that first remove the outer glucose at the C-20 position, converting Rb1 to ginsenoside Rd, followed by further deglycosylation at the C-3 position to yield compound K (CK) and Rh2. This microbial metabolism enhances the absorption of these lipophilic metabolites, as native ginsenosides have poor intestinal permeability due to their hydrophilic glycosidic structures. Following absorption, ginsenosides and their metabolites are subject to phase I and II hepatic metabolism. enzymes, particularly , facilitate oxidation of protopanaxatriol ()-type ginsenosides like Rg1, leading to derivatives such as Rh1 through or demethylation at the C-20 position. Phase II conjugation reactions, including and sulfation, further modify these compounds, often reducing their by promoting renal or biliary excretion. For instance, CK undergoes in the liver, forming CK-glucuronide, which limits its systemic exposure. Pharmacokinetic studies reveal low oral for major ginsenosides, with Rb1 exhibiting less than 5% absorption due to (P-gp) efflux in the intestines and extensive first-pass . Metabolites like CK display longer half-lives of 10-20 hours and higher concentrations compared to parent compounds. Species differences influence ginsenoside , particularly in PPD-type processing. Humans exhibit higher and levels of CK compared to rats due to distinct microbial compositions in the gut. In humans, CK is the predominant metabolite from Rb1, reflecting greater β-glucosidase activity from . Key metabolites like CK have defined structures resulting from complete deglycosylation; CK is identified as 20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol, retaining a single glucose at C-20 after removal of the outer sugars from Rb1. This structure confers improved and relative to the tetra-glycosylated parent compound.

Pharmacological Effects

Major Biological Activities

Ginsenosides exhibit a wide range of therapeutic effects, particularly in , where they demonstrate anti-tumor activity by inhibiting proliferation and . For instance, ginsenoside Rg3 has been shown to suppress epithelial-mesenchymal and invasion in cells, thereby reducing . Additionally, Rg3-enriched extracts attenuate inflammatory lesions in non-alcoholic (NAFLD) models. Other ginsenosides, such as Rh2, induce in and cells, contributing to overall tumor suppression. In cardiovascular and inflammatory contexts, ginsenosides provide cardioprotective and benefits. Ginsenoside Rb1 ameliorates ischemia-reperfusion in myocardial models by preserving cardiac and reducing . Rb1 also exerts effects in models, such as monoiodoacetate-induced in rats, by protecting and limiting joint destruction. Neuroprotective properties are evident with ginsenoside Rg1, which reduces β-amyloid levels and mitigates amyloid-beta-induced in models. Antioxidant, antidiabetic, and immunomodulatory activities further underscore the versatility of ginsenosides. Ginsenoside Re scavenges (ROS) and inhibits ROS-mediated in conditions. Rb1 enhances insulin sensitivity and , improving metabolic outcomes in diabetic models. Rg1 boosts natural killer () cell and augments immune responses against pathogens. Mixed ginsenosides from black ginseng extracts alleviate fatigue and enhance endurance in exercise-induced models by supporting energy . Structural differences influence activity profiles: protopanaxadiol (PPD)-type ginsenosides, such as Rb1, predominantly exhibit and antidiabetic effects, while protopanaxatriol ()-type, like Rg1, are more associated with neuroprotective and anti-fatigue properties. Metabolites like compound K can amplify certain activities, such as anti-tumor effects in hepatic and cancers.

Mechanisms of Action and Clinical Evidence

Ginsenosides exert their pharmacological effects through diverse molecular mechanisms, often targeting key signaling pathways involved in cellular processes such as , , and . For instance, ginsenoside Rg3 induces anti-tumor activity by promoting in cancer cells through downregulation of the anti-apoptotic protein and upregulation of pro-apoptotic proteins like Bax and cleaved caspase-3. Similarly, ginsenoside Rb1 demonstrates effects by inhibiting the expression of (COX-2) and inducible (iNOS) in activated macrophages, thereby reducing the production of proinflammatory mediators like and . In , ginsenoside Rg1 activates the PI3K/Akt signaling pathway to enhance neuronal survival and promote cerebral , which mitigates cognitive impairments in models of ischemic and . For cardiovascular benefits, ginsenoside Re upregulates endothelial (eNOS) expression via receptor-mediated PI3K/Akt activation, leading to increased production and improved endothelial function. Additionally, ginsenoside Rb1 contributes to antidiabetic effects by activating (PPARγ), which enhances , insulin sensitivity, and in adipocytes. Ginsenoside Rg5 supports defenses by acting as an of the Nrf2 , promoting its nuclear translocation and subsequent upregulation of enzymes like oxygenase-1 to counteract . Clinical evidence for ginsenosides remains promising but limited by small-scale studies and variability in formulations. A 2021 preclinical of ginsenoside Rg1 in models demonstrated significant improvements in cognitive behavioral impairments across multiple tests, with effect sizes indicating robust ; however, human RCTs are needed to confirm these findings. For ginsenoside Rg3 in cancer, a 2024 study reported its enhancement of 5-fluorouracil efficacy in cells by reducing drug resistance and Hedgehog pathway activation, while clinical observations suggest improved leukocyte counts and in patients. As of 2025, a found supplementation improved endothelial function and in humans, while a study on Rg1 in carotid patients reported reduced and better . A 2025 of animal studies highlighted ginsenosides' hepatoprotective role in NAFLD, with a mean difference in ALT levels of approximately -30 U/L, and Rg3 contributing through inhibition of and . Low oral of ginsenosides, often below 5%, poses a challenge, but nano-formulations such as liposomes and polymeric nanoparticles have shown up to 10-fold increases in absorption and targeted delivery in preclinical models. Safety profiles indicate low , with LD50 values exceeding 5 g/kg in models, supporting its general tolerability. Nonetheless, interactions with have been documented, where reduces its anticoagulant effects by upregulating hepatic enzymes, potentially increasing risk. Despite these advances, gaps persist, including the need for large-scale, randomized controlled trials comparing purified ginsenosides to whole extracts to clarify dose-response relationships and long-term efficacy.

Sources and Production

Natural Sources

Ginsenosides are triterpenoid primarily produced in species of the Panax within the family. The principal natural sources are C.A. Meyer (Asian ginseng), Panax quinquefolius L. (American ginseng), and Panax notoginseng (Burkill) F.H. Chen (Sanqi ginseng). These species are native to for P. ginseng and P. notoginseng, and eastern for P. quinquefolius. In P. ginseng roots, total ginsenoside content is approximately 4% to 6% of dry weight, with major protopanaxadiol-type (e.g., Rb1 at approximately 15 mg/g) and protopanaxatriol-type (e.g., Rg1 at approximately 15 mg/g) compounds dominating. P. quinquefolius roots exhibit comparable total levels but feature a lower Rg1:Rb1 ratio of about 0.15, indicating higher relative Rb1 content. P. notoginseng roots contain elevated total ginsenosides at around 90 mg/g dry weight, including high notoginsenoside R1 at approximately 11 mg/g. Ginsenoside concentrations differ markedly by plant part, with showing the highest levels at 1–3% dry weight, leaves and stems at 0.5–1%, and flowers and seeds containing only trace amounts. Accumulation is age-dependent, reaching a peak in of 4- to 6-year-old before stabilizing or slightly declining. Beyond the genus, minor related compounds occur in (Siberian ginseng), but these are eleutherosides rather than true dammarane-type ginsenosides. Cultivation occurs mainly in native Asian and North American regions, where wild Panax populations display greater ginsenoside structural diversity and often higher contents than cultivated counterparts grown under controlled conditions. Intensive wild harvesting poses sustainability risks, contributing to population declines and regulatory protections, such as CITES Appendix II for American ginseng (P. quinquefolius) and the Russian population of P. ginseng. Extraction from natural sources focuses on dried roots, typically employing water or (e.g., 50% ) as solvents via methods like heat-reflux or ultrasound-assisted to yield crude concentrates. Commercial supplements are standardized to exceed 5 mg/g total ginsenosides for consistency.

Biotechnological Production Methods

Plant cell and techniques have been widely employed to produce ginsenosides in controlled environments, bypassing the limitations of field cultivation such as long growth cycles and environmental variability. Hairy root cultures, induced via rhizogenes transformation, represent a prominent method for and related species, enabling stable, hormone-independent growth with enhanced accumulation. For instance, hairy root cultures of P. ginseng treated with elicitors have achieved total ginsenoside yields of up to 58.65 mg/g dry weight, with specific increases in protopanaxadiol-type ginsenosides like Rb1. optimization, such as at 5.0 mg/L, has been shown to boost overall ginsenoside production by 2-3 fold in adventitious root cultures of P. quinquefolius, reaching 105.74 mg/g from a baseline of 30.19 mg/g. Recent advances include callus-meristematic cell () cultures of , which, mediated by jasmonates, increase ginsenoside levels for sustainable production (as of November 2025). Microbial engineering offers scalable alternatives for ginsenoside synthesis by heterologously expressing plant-derived genes in host organisms. has emerged as a key chassis, particularly through integration of dammarenediol-II synthase (PgDDS) and enzyme CYP716A12, which catalyze the formation of protopanaxadiol (PPD), a core aglycone for many ginsenosides. Advances from 2015 to 2023 have enabled PPD production exceeding 500 mg/L in shake flasks and up to 11 g/L in fed-batch fermentations, with further pathways yielding bioactive derivatives like Rh2 at 2.25 g/L. Enzymatic provides a targeted approach to convert abundant major ginsenosides into rare, more bioavailable forms like compound K (CK). Glycosidases from species, such as β-glucosidases, facilitate deglycosylation of protopanaxadiol-type ginsenosides (e.g., Rb1 to CK) with high , often exceeding 90% conversion on industrial scales through sequential . Conversely, glycosyltransferases (UGTs) enable regioselective of aglycones like PPD to synthesize specific ginsenosides, with -derived enzymes demonstrating robust activity in bioprocesses for CK production. Recent innovations in and have further refined ginsenoside production. CRISPR-Cas9-mediated knockout of dammarenediol-II synthase in P. ginseng has altered saponin profiles, increasing protopanaxadiol-type ginsenosides by up to 50% relative to protopanaxatriol types in regenerated plants, as reported in 2022-2023 studies targeting pathway flux. platforms, including E. coli cell factories expressing multi-enzyme cascades, have achieved CK titers of up to several grams per liter through pathway optimization and cofactor engineering, highlighted in 2025 reviews on sustainable production. These approaches leverage natural plant enzymes like CYP716A12 for in microbial hosts. Biotechnological methods provide advantages in , batch-to-batch , and customizable ginsenoside profiles compared to variable extracts, potentially reducing reliance on wild harvesting. However, challenges persist, including regulatory approvals for genetically modified products and optimization of upstream precursors to achieve commercial viability.

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