Search Results (17)

Ginkgo biloba seeds are a rich source of flavonoids and the unique terpene lactones—ginkgolides and bilobalide, known for their neuroprotective and cognitive-improving effects. However, unlike the widely used leaves, the seeds contain substantial levels of ginkgolic acid and ginkgotoxin (4′-O-methylpyridoxine), an antivitamin B₆ compound. At high concentrations, ginkgotoxin exhibits neurotoxicity, potentially inducing seizures, respiratory distress, and loss of consciousness, thus limiting the safe application of Ginkgo seed-derived products. This study aimed to develop a simple yet effective extraction protocol that reduces ginkgotoxin levels in Ginkgo biloba seed extracts while preserving their beneficial phytochemicals. A multistep liquid–liquid extraction approach employing sequential polar and non-polar solvents was implemented. Following each extraction stage, fractions were analyzed using ultra-high-performance liquid chromatography coupled with mass spectrometry (UHPLC–MS). The concentrations of flavonoids, ginkgolides, bilobalide, ginkgolic acid, and ginkgotoxin were quantified to evaluate detoxification efficiency and phytochemical retention. Compared with conventional single-step extraction using 70% methanol, this multistep protocol markedly reduced ginkgotoxin and ginkgolic acid to near-undetectable levels, while preserving detectable concentrations of major flavonoids and terpene trilactones. The findings demonstrate that multistep extraction represents a promising and practical strategy for minimizing ginkgotoxin in Ginkgo biloba seed extracts without compromising their beneficial phytochemical composition. This approach provides a sound basis for developing safer, functionally active Ginkgo-based products. Full article

Temperature is a major determinant of plant metabolic plasticity, yet its role in directing volatile and semi-volatile specialized metabolism in Ginkgo biloba remains poorly understood. In this study, we investigated how contrasting low- and high-temperature treatments reshape secondary metabolite contents in G. biloba microclones cultivated in vitro. Plants were exposed to cold (+3 °C) and heat (+30 °C) conditions, and their responses were analyzed using GC–MS profiling, anatomical measurements, chlorophyll fluorescence, and multivariate statistics. Cold treatment selectively increased the abundances of monoterpenes (13.22%) and sesquiterpenes (13.83%), with the strongest accumulation of caryophyllene, eucalyptol, and (1S)-camphor. In contrast, heat treatment reduced ester content to 3.73% and strongly enriched oxy-sesquiterpenes (46.50%) and lactone/ketone/spiroketone (29.54%) contents. The enhanced accumulation of isocalamendiol, isoshyobunone, cyclohexanone derivative, dehydroxy-isocalamendiol, and (+)-2-bornanone was observed under heat. According to the multivariate analysis, control plants were associated with traits reflecting optimal physiological performance, including greater parenchyma, phloem, and xylem thickness, larger vascular bundles, longer stomata, and higher NPQ, qN, Y(NPQ), and F_v/F_m. Cold-treated plants showed thicker epidermis and sclerenchyma, higher stomatal density and width, elevated Y(NO), and an enrichment of esters and terpenoids, whereas heat-treated plants were characterized by thicker adaxial and abaxial epidermis, increased mesophyll thickness, and higher levels of oxygenated metabolites. These findings expand current knowledge beyond terpene trilactones and flavonoids and identify Ginkgo microclones as a useful in vitro model for temperature-guided metabolic reprogramming and targeted metabolite enrichment. Full article

Ginkgo biloba is an ancient plant that has survived up until the present day. Gingko biloba is a rich source of valuable secondary metabolites, particularly terpene trilactones (TTLs) such as ginkgolides and bilobalides, which are obtained from the leaves and seeds of the plant. TTLs have pharmacological properties, including anticancer, anti-dementia, antidepressant, antidiabetic, anti-inflammatory, anti-hypertensive, antiplatelet, immunomodulatory, and neuroprotective effects. However, ginkgo is a very-slow-growing tree that takes approximately 30 years to reach maturity. In addition, the accumulation of TTLs in these plants is affected by age, sex, and seasonal and geographical variations. Therefore, plant cell cultures have been established in ginkgo to produce TTLs. Extensive investigations have been conducted to optimize the culture media, growth regulators, nutrients, immobilization, elicitation, and precursor-feeding strategies for the production of TTLs in vitro. In addition, metabolic engineering and synthetic biology methods have been used for the heterologous production of TTLs. In this review, we present the research strategies applied to cell cultures for the production of TTLs. Full article

Ginkgo biloba L. (ginkgo) is a widely used medicinal plant around the world. Its leaves, which have been used as a traditional Chinese medicine, are rich in various bioactive components. However, most of the research and applications of ginkgo leaves have focused on terpene trilactones and flavonol glycosides, thereby overlooking the other active components. In this study, a lipophilic extract (GL) was isolated from ginkgo leaves. This extract is abundant in lipids and lipid-like molecules. Then, its effect and potential mechanism on glucose uptake and insulin resistance in C2C12 myotubes were investigated. The results showed that GL significantly enhanced the translocation of GLUT4 to the plasma membrane, which subsequently promoted glucose uptake. Meanwhile, it increased the phosphorylation of AMP-activated protein kinase (AMPK) and its downstream targets. Both knockdown of AMPK with siRNA and inhibition with AMPK inhibitor compound C reversed these effects. Additionally, GL ameliorated palmitate-induced insulin resistance by enhancing insulin-stimulated glucose uptake, increasing the phosphorylation of protein kinase B (PKB/AKT), and restoring the translocation of GLUT4 from the cytoplasm to the membrane. However, pretreatment with compound C abolished these beneficial effects of GL. In conclusion, GL enhances basal glucose uptake in C2C12 myotubes and improves insulin sensitivity in palmitate-induced insulin resistant myotubes through the AMPK pathway. Full article

As a medicinal tree species, ginkgo (Ginkgo biloba L.) and terpene trilactones (TTLs) extracted from its leaves are the main pharmacologic activity constituents and important economic indicators of its value. The accumulation of TTLs is known to be affected by environmental stress, while the regulatory mechanism of environmental response mediated by microRNAs (miRNAs) at the post-transcriptional levels remains unclear. Here, we focused on grafted ginkgo grown in northwestern, southwestern, and eastern-central China and integrally analyzed RNA-seq and small RNA-seq high-throughput sequencing data as well as metabolomics data from leaf samples of ginkgo clones grown in natural environments. The content of bilobalide was highest among detected TTLs, and there was more than a twofold variation in the accumulation of bilobalide between growth conditions. Meanwhile, transcriptome analysis found significant differences in the expression of 19 TTL-related genes among ginkgo leaves from different environments. Small RNA sequencing and analysis showed that 62 of the 521 miRNAs identified were differentially expressed among different samples, especially the expression of miRN50, miR169h/i, and miR169e was susceptible to environmental changes. Further, we found that transcription factors (ERF, MYB, C3H, HD-ZIP, HSF, and NAC) and miRNAs (miR319e/f, miRN2, miRN54, miR157, miR185, and miRN188) could activate or inhibit the expression of TTL-related genes to participate in the regulation of terpene trilactones biosynthesis in ginkgo leaves by weighted gene co-regulatory network analysis. Our findings provide new insights into the understanding of the regulatory mechanism of TTL biosynthesis but also lay the foundation for ginkgo leaves’ medicinal value improvement under global change. Full article

Ginkgo biloba is a relict tree species showing high resistance to adverse biotic and abiotic environmental factors. Its fruits and leaves have high medicinal value due to the presence of flavonoids, terpene trilactones and phenolic compounds. However, ginkgo seeds contain toxic and allergenic alkylphenols. The publication revises the latest research results (mainly from 2018–2022) regarding the chemical composition of extracts obtained from this plant and provides information on the use of extracts or their selected ingredients in medicine and food production. A very important section of the publication is the part in which the results of the review of patents concerning the use of Ginkgo biloba and its selected ingredients in food production are presented. Despite the constantly growing number of studies on its toxicity and interactions with synthetic drugs, its health-promoting properties are the reason for the interest of scientists and motivation to create new food products. Full article

The phytochemical profiles of extracts from closed, semi-opened and opened leaf buds and the summer leaves of Ginkgo biloba were studied. The extraction and purification of bilobalide and ginkgolides, using andrographolide as an internal standard, were optimised. The terpene trilactone concentrations increased with bud development, from 1.07 mg/g dry wt in closed buds to a maximum of 3.75 mg/g dry wt in summer leaves. The major terpene trilactone was bilobalide at all developmental stages. The concentration of flavonol aglycones in hydrolysed extracts was also analysed. The flavonol glycoside concentration increased from the closed bud stage (0.21 ± 0.01% dry wt) to the summer leaf stage (1.15 ± 0.01% dry wt). A linear correlation was observed between the terpene trilactone and flavonoid content during gingko leaf development. Full article

Ginkgo tea and ginkgo wine are two familiar Ginkgo biloba leaf extract (GBE) drinks in the form of dietary supplements (DS) used for healthcare in east Asia. Nevertheless, a comprehensive evaluation of their safety and efficacy is still lacking. In this study, GBE drinks were prepared from naturally newly senescent yellow leaves (YL) and green leaves (GL) in autumn. Their total flavonoids, antioxidant capacity and prescribed ingredients were investigated. In brief, the proportions of total flavonoids, total flavonol glycosides (TFs), total terpene trilactones (TTLs) and ginkgolic acids in the GBE drinks all did not meet the standards of worldwide pharmacopoeias. Specifically, the levels of TFs in the ginkgo tea prepared from YL were significantly higher than that prepared from GL. Further analyses revealed a substandard ratio of isorhamnetin/quercetin and an accumulation of leaf-age-related compounds, which were both unqualified. The proportions of specific TTLs varied between the ginkgo tea and ginkgo wine, although no significant differences were detected in terms of the total levels of TTLs. Noticeably, numerous biflavones and thousands of times over the limiting concentration of ginkgolic acids, including newly identified types, were only detected in ginkgo wine. Finally, the use of the GBE drinks as DSs was comprehensively evaluated according to the acceptable daily intake. This study showed the limited healthcare effects of GBE drinks despite their powerful antioxidant capacity. Full article

Ginkgo (Ginkgo biloba L.) is one of the most distinctive plants, characterized by excellent resistance to various environmental conditions. It is used as an ornamental plant and is recognized as a medicinal plant in both traditional and Western medicine. Its bioactive potential is associated with the presence of flavonoids and terpene trilactones, but many other compounds may also have synergistic effects. Flavonoid dimers—biflavonoids—are important constituents of ginkgophytopharmaceuticals. Currently, the presence of 13 biflavonoids has been reported in ginkgo, of which amentoflavone, bilobetin, sciadopitysin, ginkgetin and isoginkgetin are the most common. Their role in plants remains unknown, but their bioactivity and potential role in the management of human health are better investigated. In this review, we have provided an overview of the chemistry, diversity and biological factors that influence the presence of biflavonoids in ginkgo, as well as their bioactive and health-related properties. We have focused on their antioxidant, anticancer, antiviral, antibacterial, antifungal and anti-inflammatory activities as well as their potential role in the treatment of cardiovascular, metabolic and neurodegenerative diseases. We also highlighted their potential toxicity and pointed out further research directions. Full article

Terpene trilactones (TTLs) are the main secondary metabolites in Ginkgo biloba L. with efficacious pharmacological activity. Jasmonate ZIM-domain (JAZ) protein is a key regulatory factor of the JA signaling pathway, which regulates the biosynthesis of secondary metabolites such as terpenes, alkaloids, and flavonoids. In this study, GbJAZ01~GbJAZ11 were identified from the genome data in G. biloba, which contained TIFY-, Jas-, and weakly conserved NT-domains, and the promoters in most of them contained light, hormone, and stress-responsive elements. Phylogenetic analysis divided all JAZ proteins of Arabidopsis thaliana, Oryza sativa, Picea sitchensis, Taxus chinensis, and G. biloba into nine groups, in which GbJAZs belong to Group VI-IX. GbJAZs have similar functional motifs to A. thaliana and O. sativa, but also contain three specific motifs of gymnosperms, indicating that, although gymnosperms and angiosperms have some conservative structures and functions, their evolutionary processes are independent. Expression pattern analysis showed that the expression levels of GbJAZs were significantly up-regulated by MeJA, but the change pattern and amplitude were different, indicating that the function of GbJAZs in response to a JA signal may be different. After ABA and SA treatment, the expression of GbJAZs was up-regulated or inhibited in varying degrees, and different GbJAZs may be involved in the synergistic or antagonistic effects between JA and other hormone signals. The MeJA significantly increased the content of TTLs in G. biloba leaves, which were significantly positively correlated with the expression levels of GbJAZ01, 02, 07, and 11, and negatively correlated with the expression of GbJAZ04. They may play an important role in JA signaling pathways and the interactions between JA and other hormone signals, and participate in the regulation of the biosynthesis of TTLs. Our results provide a reference for the discovery that GbJAZs are involved in JA signaling pathways, and lay a theoretical foundation for analyzing JA signaling pathways to regulate the synthesis of secondary metabolites. Full article

The present study compared the effects of natural senescence and methyl jasmonate (JA-Me) treatment on the levels of terpene trilactones (TTLs; ginkgolides and bilobalide), phenolic acids, and flavonoids in the primary organs of Ginkgo biloba leaves, leaf blades, and petioles. Levels of the major TTLs, ginkgolides B and C, were significantly higher in the leaf blades of naturally senesced yellow leaves harvested on 20 October compared with green leaves harvested on 9 September. In petioles, a similar effect was found, although the levels of these compounds were almost half as high. These facts indicate the importance of the senescence process on TTL accumulation. Some flavonoids and phenolic acids also showed changes in content related to maturation or senescence. Generally, the application of JA-Me slightly but substantially increased the levels of TTLs in leaf blades irrespective of the difference in its application side on the leaves. Of the flavonoids analyzed, levels of quercetin, rutin, quercetin-4-glucoside, apigenin, and luteolin were dependent on the JA-Me application site, whereas levels of (+) catechin and (−) epicatechin were not. Application of JA-Me increased ferulic acid and p-coumaric acid esters in the petiole but decreased the levels of these compounds in the leaf blade. The content of p-coumaric acid glycosides and caffeic acid esters was only slightly modified by JA-Me. In general, JA-Me application affected leaf senescence by modifying the accumulation of ginkogolides, flavonoids, and phenolic acids. These effects were also found to be different in leaf blades and petioles. Based on JA-Me- and aging-related metabolic changes in endogenous levels of the secondary metabolites in G. biloba leaves, we discussed the results of study in the context of basic research and possible practical application. Full article

Ginkgo biloba has been cultivated in Bulgaria since the end of the 19th century. Ividual specimens can be seen in almost every park. Females of the tree are considered contaminants of the landscape because their ripe seeds have a strong odor and are not utilized. We undertook this study to clarify whether ginkgo seeds of local origin can be converted from an unwanted and unused environmental pollutant into a source of beneficial compounds. Various analytical and chromatographic methods were used to quantify the major constituents and ten biologically active compounds in methanol seed extract. The results showed that the seeds are low in proteins (5%) and fats (1%); the seeds were also rich in unsaturated fatty acids and tocopherols. About 44% of nut starch was resistant to in vitro enzymatic hydrolysis. The amount of terpene trilactones in an aqueous-methanol seed extract was significantly higher than the number of flavonoids. Ginkgotoxin and ginkgolic acid were also found. The extract demonstrated weak antimicrobial activity against thirteen microorganisms. This study revealed that seeds of locally grown Ginkgo trees can be used as a source of biologically active substances. The chemical composition show similarity to those of seeds from other geographical areas. Full article

Ginkgo biloba leaves have various health benefits due to the presence of bioactive compounds such as polyprenyl acetates, flavonoids, and terpene trilactones. However, there is little literature reported on the aromatic acids in Ginkgo biloba leaves. In this work, five aromatic acids including shikimic acid (SA), 6-hydroxykynurenic acid (6-HKA), protocatechuic acid (PA), gallic acid (GAA), and p-hydroxybenzoic acid (PHBA) were simultaneously extracted from Ginkgo biloba leaves by employing the green deep eutectic solvents (DESs). A DES tailor-made from xylitol, glycolic acid and 1,5-pentanedioic acid at a molar ratio of 1:3:1 with 50% (w/w) water addition, named as NGG50, gave higher extraction yields for the five aromatic acids. Main factors affecting the extraction process were further optimized. The highest extraction yields of SA, GAA, 6-HKA, PA, and PHBA were 94.15 ± 0.96 mg/g, 332.69 ± 5.19 μg/g, 25.90 ± 0.61 μg/g, 429.89 ± 11.47 μg/g and 67.94 ± 0.37 μg/g, respectively. The NGG50-based extraction process developed here was a successful attempt of simultaneously extracting five aromatic acids from Ginkgo biloba leaves for the first time, which could provide a new exploitation direction of Ginkgo biloba leaves. Full article

3-Hydroxy-3-methylglutaryl-CoA synthase (HMGS) is one of the rate-limiting enzymes in the mevalonate pathway as it catalyzes the condensation of acetoacetyl-CoA to form 3-hydroxy-3-methylglutaryl-CoA. In this study, A HMGS gene (designated as GbHMGS1) was cloned from Ginkgo biloba for the first time. GbHMGS1 contained a 1422-bp open-reading frame encoding 474 amino acids. Comparative and bioinformatics analysis revealed that GbHMGS1 was extensively homologous to HMGSs from other plant species. Phylogenetic analysis indicated that the GbHMGS1 belonged to the plant HMGS superfamily, sharing a common evolutionary ancestor with other HMGSs, and had a further relationship with other gymnosperm species. The yeast complement assay of GbHMGS1 in HMGS-deficient Saccharomyces cerevisiae strain YSC6274 demonstrated that GbHMGS1 gene encodes a functional HMGS enzyme. The recombinant protein of GbHMGS1 was successfully expressed in E. coli. The in vitro enzyme activity assay showed that the k_cat and K_m values of GbHMGS1 were 195.4 min⁻¹ and 689 μM, respectively. GbHMGS1 was constitutively expressed in all tested tissues, including the roots, stems, leaves, female flowers, male flowers and fruits. The transcript accumulation for GbHMGS1 was highest in the leaves. Expression profiling analyses revealed that GbHMGS1 expression was induced by abiotic stresses (ultraviolet B and cold) and hormone treatments (salicylic acid, methyl jasmonate, and ethephon) in G. biloba, indicating that GbHMGS1 gene was involved in the response to environmental stresses and plant hormones. Full article

Ginkgolides and bilobalide, collectively termed terpene trilactones (TTLs), are terpenoids that form the main active substance of Ginkgo biloba. Terpenoids in the mevalonate (MVA) biosynthetic pathway include acetyl-CoA C-acetyltransferase (AACT) and mevalonate kinase (MVK) as core enzymes. In this study, two full-length (cDNAs) encoding AACT (GbAACT, GenBank Accession No. KX904942) and MVK (GbMVK, GenBank Accession No. KX904944) were cloned from G. biloba. The deduced GbAACT and GbMVK proteins contain 404 and 396 amino acids with the corresponding
open-reading frame (ORF) sizes of 1215 bp and 1194 bp, respectively. Tissue expression pattern analysis revealed that GbAACT was highly expressed in ginkgo fruits and leaves, and GbMVK was highly expressed in leaves and roots. The functional complementation of GbAACT in AACT-deficient Saccharomyces cerevisiae strain Δerg10 and GbMVK in MVK-deficient strain Δerg12 confirmed that GbAACT mediated the conversion of mevalonate acetyl-CoA to acetoacetyl-CoA and GbMVK mediated the conversion of mevalonate to mevalonate phosphate. This observation indicated that GbAACT and GbMVK are functional genes in the cytosolic mevalonate (MVA) biosynthesis pathway. After G. biloba seedlings were treated with methyl jasmonate and salicylic acid, the expression levels of GbAACT and GbMVK increased, and TTL production was enhanced. The cloning, characterization, expression and functional analysis of GbAACT and GbMVK will be helpful to understand more about the role of these two genes involved in TTL biosynthesis. Full article