Search Results (51)

α-Bisabolol is naturally occurring in many plants and has great potential in health products and pharmaceuticals. However, the current extraction method from natural plants is unsustainable and cannot fulfil the increasing requirement. This study aimed to develop a sustainable strategy to enhance the biosynthesis of α-bisabolol by metabolic engineering. Integration of ERG20 (encoding farnesyl diphosphate synthase) and tHMG1 (encoding truncated 3-hydroxy-3-methylglutaryl-CoA reductase) genes with a constitutive strong promoter into the yeast genome elevated α-bisabolol production from 20.21 mg/L to 98.30 mg/L, representing a 4.86-fold increase. Further optimization of the mevalonate pathway and amplification of ERG20, tHMG1, and OsTPS1 copy numbers enhanced α-bisabolol synthesis to 423.01 mg/L, achieving a 20.93-fold improvement relative to the baseline. This work establishes a reference strategy for high-yield α-bisabolol biosynthesis in engineered yeast. Full article

The uterus is a dynamic organ in which the endometrium undergoes cyclic processes of proliferation, shedding, and regeneration under the influence of estrogen and progesterone. In particular, estrogen regulates the proliferation and differentiation of the endometrium and plays an important role in the development of gynecological diseases such as endometrial cancer. Farnesyl diphosphate synthase (FDPS) is a key enzyme involved in the mevalonate pathway, catalyzing the synthesis of farnesyl pyrophosphate (FPP), which plays an essential role in cholesterol biosynthesis and protein prenylation. In this study, we demonstrated using an in vivo mouse model that the expression of FDPS is regulated by estrogen. FDPS expression was specifically elevated during the proestrus stage of the estrous cycle and subsequently decreased. In ovariectomized (OVX) mice, FDPS expression was significantly increased 24 h after estrogen treatment, whereas this response was suppressed by treatment with the estrogen receptor alpha (ERα) antagonist, ICI 182,780. Although FDPS expression has been reported in various cancers, its role in endometrial cancer remains unclear. Histological and cellular analyses revealed that FDPS is highly expressed in human endometrial cancer tissues and in the endometrial cancer cell line Ishikawa, where it contributes to cell proliferation. These findings suggest that FDPS may play a role in the survival and growth of endometrial cancer cells. This study provides new insights into the potential function of FDPS in the uterus and suggests that targeting FDPS may represent a promising therapeutic strategy for endometrial cancer. Full article

Floral volatile terpenoids are known to play important roles in plant pollination biology by attracting animal pollinators, repelling antagonists, and enhancing resistance to potential microbial pathogens. The terpenoid blend emitted by a flower is usually plant-lineage specific and is primarily determined by a set of versatile terpene synthases (TPSs), which catalyze the final step of diverse terpenoid synthesis. The strongly scented flower of Jasminum sambac (L.) Aiton emits linalool and α-farnesene, which dominate the nocturnal floral VOCs, yet the corresponding TPSs have not been identified. Here, we show that four TPS enzymes are responsible for the synthesis of a mixture of volatile terpenoids in the flower, based on their highly correlated and almost exclusive expression in the petal, as well as their enzymatic characterizations in vitro and in Nicotiana benthamiana Domin. JsTPS01 (TPS-a) acts as a sesquiterpene synthase, producing τ-cadinol in yeast at levels that mirror its rhythmic expression in petals. JsTPS02 (TPS-b) carries a plastid-targeting transit peptide, localizes to chloroplasts/plastids, and converts geranyl diphosphate (GPP) to linalool with high affinity (K_m = 28.2 ± 3.4 µM). JsTPS03 is a TPS-b clade member that can convert farnesyl diphosphate (FPP) to farnesol with a K_m of 14.4 ± 5.9 μM in an in vitro assay using isolated yeast vehicles. JsTPS04 (TPS-e/f) exhibits dual targeting—cytosolic in protoplasts of Arabidopsis thaliana (L.) Heynh, but plastidic in J. sambac petals—and functions as a bifunctional mono-/sesqui-TPS, forming linalool from GPP (K_m = 2.5 ± 0.3 µM) and trans-nerolidol from FPP (K_m = 7.6 ± 0.6 µM). Transient expression in N. benthamiana leaves further confirmed its in-planta linalool production. Collectively, we identified four preferentially expressed terpene synthases that contribute to the production of linalool, τ-cadinol, trans-nerolidol, and farnesol in J. sambac. Full article

Watercore disease, a physiological disorder in pineapple (Ananas comosus), manifests during late fruit development. Affected fruits develop water-soaked flesh and reduced storability. (1) Background: To explore underlying molecular mechanisms, comparative proteomic profiling was conducted in this study. (2) Methods: Data-independent acquisition (DIA) strategy was employed for comparative analysis between the resistant germplasm “35-1” and the susceptible germplasm “29-3”, as well as between the healthy and diseased “Paris”. (3) Results: Resistant (“35-1”) versus susceptible (“29-3”) germplasm analysis revealed differentially expressed proteins (DEPs) and unique proteins (SEPs) enriched in cell walls, secretory vesicles, and apoplast, functioning in cell wall loosening, hormone response, isoflavonoid biosynthesis, and farnesyl diphosphate biosynthesis. Healthy versus diseased “Paris” pulp analysis showed DEPs/SEPs enrichment in ribosomal small subunit biogenesis. These proteins form a central regulatory network potentially orchestrating tRNA synthesis, tubulin biosynthesis, and other carbohydrate metabolism. Partial protein overlap occurred in germplasm- and disease-derived differences. Resistant germplasm (“35-1”) and healthy “Paris” accumulated stress-responsive/resistant proteins and cell wall-modifying enzymes (e.g., phenylalanine ammonia-lyase, raffinose synthase, expansins, and mannan hydrolase). Susceptible germplasm (“29-3”) and diseased “Paris” exhibited prominent stress-responsive protein accumulation, such as alcohol dehydrogenase, 1-aminocyclopropane-1-carboxylate oxidase, and hypoxia-induced protein. (4) Conclusions: This comparative proteomics study identifies pineapple watercore resistance/susceptibility-associated proteins, providing a molecular basis for resistant germplasm development and disorder control. Full article

Lavender species are economically valuable plants, widely cultivated for their essential oils (EOs), which include sesquiterpenes. The sesquiterpenes caryophyllene and cadinol are major constituents, contributing woody and balsamic notes. However, the specific enzymes catalyzing their formation in lavender have not been elucidated. This study reports the comprehensive functional and structural characterization of two pivotal sesquiterpene synthases from Lavandula angustifolia (lavender): caryophyllene synthase (CARS) and cadinol synthase (CADS). Mutation experiments were performed based on molecular docking predictions, revealing that negatively charged residues interact electrostatically with magnesium ions (Mg²⁺). Both deletion of 1–226 and 1–228 (∆1–226 and ∆1–228) display activity levels equivalent to their corresponding wild-type proteins, while deletions at positions 522–548 and 529–555 significantly enhanced enzyme activity. Additionally, the highest expression levels of CARS were in the flowers under white light for 8 h, while CADS exhibited peak expression in the leaves under white light for 12 h. These findings deepen our understanding of the regulatory mechanisms involved in sesquiterpene biosynthesis in lavender and provide insights for genetic engineering strategies aimed at enhancing EO production. Such advances could also inform the development of cosmetic, personal care, and medicinal products. Full article

We investigated the dynamic changes in volatile aroma compound profiles (types and concentrations) and associated gene expression patterns in both the peel and pulp tissues of apples during fruit maturation. This study aimed to elucidate the metabolic regulatory mechanisms underlying volatile aroma biosynthesis in Malus domestica “Ningqiu” apples, thereby providing theoretical support for the comprehensive utilization of aroma resources. Our methodological framework integrated headspace solid-phase microextraction gas chromatography–mass spectrometry (HS-SPME-GC-MS), ultra-high-performance liquid chromatography–orbitrap mass spectrometry (UHPLC-OE-MS), and Illumina high-throughput sequencing to generate comprehensive metabolomic and transcriptomic profiles of peel and pulp tissues. Critical differential aroma compound classes were identified, including esters, aldehydes, alcohols, terpenoids, and ketones, with their metabolic pathways systematically mapped through KEGG functional annotation. Our findings revealed substantial transcriptomic and metabolomic divergence across carotenoid, terpenoid, and fatty acid metabolic pathways. Integrative analysis of multi-omics data revealed 26 and 31 putative biologically significant hub genes in peel and pulp tissues, respectively, putatively associated with the observed metabolic signatures. Among these, five core genes—farnesyl diphosphate synthase (FDPS1.X1), alcohol acyltransferases (AAT1 and AAT3), alcohol dehydrogenase (ADH3), and carotenoid cleavage dioxygenase (CCD3)—were recognized as shared regulatory determinants between both tissue types. Furthermore, terpene synthase (TPS7) emerged as a peel-specific regulatory factor, while hydroperoxide lyase (HPL2), alcohol dehydrogenases (ADH2 and ADH4), and alcohol acyltransferase (AAT2) were identified as pulp-exclusive modulators of metabolic differentiation. The experimental findings provide foundational insights into the molecular basis of aroma profile variation in Malus domestica “Ningqiu” and establish a functional genomics framework for precision breeding initiatives targeting fruit quality optimization through transcriptional regulatory network manipulation. Full article

This publication reports the controlled cultivation of Zanthoxylum chiloperone var. angustifolium Engl. (Rutaceae) in several growth substrates under controlled greenhouse conditions. This plant is well-known for its anti-Chagas (trypanocidal) activity, related to the presence of several β-carboline alkaloids. The metabolomic study of Z. chiloperone seedlings over two years of growth (2018–2020) was performed using comprehensive two-dimensional gas chromatography time-of-flight mass spectrometry (GC × GC-TOFMS). The canthin-6-one alkaloids, canthin-6-one and 5-methoxy-canthin-6-one, were putatively identified in Z. chiloperone extracts. Finally, in vitro and in silico studies of trypanocidal activity were performed, suggesting that canthin-6-one alkaloids could interact with the main pharmacological targets against Trypanosoma cruzi, cruzain protease, dihydroorotate dehydrogenase, lanosterol 14-alpha-demethylase, farnesyl diphosphate, and squalene synthases. Full article

Ergosterol is a component of fungal cell membranes that has physiological functions and applications in drugs, such as anti-inflammatory, anti-tumor, anti-fungal, and other immunosuppressive activities. The fungus Athelia termitophila, also known as the termite ball fungus, primarily contains secondary metabolites (like active ingredients) that are similar to ergosterol. To enhance the synthesis of ergosterol and mycelial biomass in termite ball fungus, methyl jasmonate and salicylic acid were used to stimulate the biosynthesis of ergosterol compounds during the growth of TMB mycelium and relative quantitative levels of gene transcripts. The conditions of the inducers were optimized. Under 80 µmol/L MJ incubation conditions, the content of ergosterol compounds in TMB was increased by 2.23-fold compared with the wild-type strain. RT-qPCR results at the transcriptional level of ergosterol synthesis pathway genes showed that MJ significantly induced the expression of HMGR (3-Hydroxy-3-Methylglutaryl-Coa Reductase), HMGS (3-Hydroxy-3-Methylglutaryl-Coa Synthase), SE (Squalene Epoxidase), and FPS (Farnesyl Diphosphate Synthase) genes in the ergosterol synthesis pathway. For expression levels at different induction days, we collected 7/10 d and 4/6/8 d samples with similar expression patterns, as well as SS (Squalene Synthase)/FPS (Farnesyl Diphosphate Synthase), SE (Squalene Epoxidase)/MVD (Mevalonate Diphosphate Decarboxylase), and HMGS (3-Hydroxy-3-Methylglutaryl-Coa Synthase)/HMGR (3-Hydroxy-3-Methylglutaryl-Coa Reductase) genes with similar expression patterns, which resulted in gene transcription data during ergosterol content synthesis. The current study provides an effective method to increase the ergosterol contents in termite ball fungus and a new idea for the mechanism of MJ-induced ergosterol compound biosynthesis. Full article

The biosynthesis of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP), which are essential for sesquiterpenes and triterpenes, respectively, is primarily governed by the mevalonate pathway, wherein farnesyl pyrophosphate synthase (FPS) plays a pivotal role. This study identified eight members of the FPS gene family in Euphorbia hirta, designated EhFPS1–EhFPS8, through bioinformatics analysis, revealing their distribution across several chromosomes and a notable tandem gene cluster. The genes exhibited strong hydrophilic properties and key functional motifs crucial for enzyme activity. An in-depth analysis of the EhFPS genes highlighted their significant involvement in isoprenoid metabolism and lipid biosynthesis, with expression patterns influenced by hormones such as jasmonic acid and salicylic acid. Tissue-specific analysis demonstrated that certain FPS genes, particularly EhFPS1, EhFPS2, and EhFPS7, showed elevated expression levels in latex, suggesting their critical roles in terpenoid biosynthesis. Furthermore, subcellular localization studies have indicated that these proteins are primarily found in the cytoplasm, reinforcing their function in metabolic processes. These findings provide a foundational understanding of the FPS genes in E. hirta, including their gene structures, conserved domains, and evolutionary relationships. This study elucidates the potential roles of these genes in response to environmental factors, hormone signaling, and stress adaptation, thereby paving the way for future functional analyses aimed at exploring the regulation of terpenoid biosynthesis and enhancing stress tolerance in this species. Full article

The development of chicken ovarian follicles involves two key stages of primordial follicle recruitment and follicle selection that are tightly regulated by multiple reproductive hormones and cytokines. Our previous study revealed an estrogen-stimulated increase in the phosphorylation level of serine at position 54 of lysine demethylase 1A (LSD1Ser54p) in the theca cells of chicken hierarchical ovarian follicles (Post-TCs). In this study, we further found that the upregulation of LSD1Ser54p by estrogen was performed by glycogen synthase kinase 3 beta (GSK3β) and that GSK3β promoted LSD1Ser54p levels by directly binding to the SWIRM and AOL1 domains of LSD1. Upon estrogen stimulation, the phosphorylation level of tyrosine at position 216 of GSK3β (GSK3βTyr216p) increased, which enhanced the binding between LSD1 and GSK3β. The subsequent transcriptome sequencing on chicken Post-TCs treated with estrogen and CUT&RUN sequencing against the LSD1Ser54p protein revealed that the expression of the farnesyl-diphosphate farnesyltransferase 1 (FDFT1) gene was simultaneously upregulated by estrogen, GSK3β, and LSD1Ser54p. Moreover, the overexpression of FDFT1 further promoted cholesterol biosynthesis in chicken Post-TCs. In short, the findings of this study suggest that estrogen-induced tyrosine phosphorylation at position 216 of GSK3β can upregulate the level of LSD1Ser54p, leading to the activation of FDFT1 expression and subsequently promoting cholesterol biosynthesis in chicken Post-TCs, which may in turn enhance estrogen synthesis. Full article

Construction of the genome of Hericium coralloides, a species of edible mushroom, and identification of the genes involved in terpenoid biosynthesis can determine the biology and genetics of terpenoids. The present study describes the assembly of a high-quality chromosome-scale genome of H. coralloides using Pacbio HiFi sequencing and Hi-C technology. This genome consisted of 13 chromosomes, a total size of 43.6 Mb, contigs of N50 3.6 Mb, GC content at 54%, and BUSCOs integrity of 96.9%. Genes associated with terpenoid biosynthesis were predicted by KEGG enrichment analysis and homologous alignment. The Her011461 and Her008335 genes, encoding proteins in the terpenoid backbone synthesis pathway, were found to encode geranylgeranyl pyrophosphate and farnesyl diphosphate synthases, key enzymes in the biosynthesis of geranylgeranyl diphosphate, a precursor of several diterpenoids. Her011463 was found to be involved in regulating diterpene cyclase. The Her005433, Her006724, Her010605, and Her010608 genes were found to encode sesquiterpene synthesis. Most of these genes were more highly expressed in dikaryotic mycelia than in the primordium and fruiting bodies, indicating that terpenoids may be more abundant in dikaryotic mycelia. To our knowledge, this study is the first to assemble the H. coralloides genome at the chromosome scale and to identify the genes involved in terpenoid biosynthesis. Full article

Nanoscale geranium waste (GW) and magnesium nanoparticle/GW nanocomposites (Mg NP/GW) were prepared using green synthesis. The Mg NP/GW samples were subjected to characterization using X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR-FT). The surface morphology of the materials was examined using a scanning electron microscope (SEM), and their thermal stability was assessed through thermal gravimetric analysis (TG). The BET-specific surface area, pore volume, and pore size distribution of the prepared materials were determined using the N2 adsorption–desorption method. Additionally, the particle size and zeta potentials of the materials were also measured. The influence of the prepared nanomaterials on seed germination was intensively investigated. The results revealed an increase in seed germination percent at low concentrations of Mg NP/GWs. Upon treatment with Mg NP/GW nanoparticles, a reduction in the mitotic index (MI) was observed, indicating a decrease in cell division. Additionally, an increase in chromosomal abnormalities was detected. The efficacy of GW and Mg NP/GW nanoparticles as new elicitors was evaluated by studying their impact on the expression levels of the farnesyl diphosphate synthase (FPPS1) and geranylgeranyl pyrophosphate (GPPS1) genes. These genes play a crucial role in the terpenoid biosynthesis pathway in Sinapis alba (S. alba) and Pelargonium graveolens (P. graveolens) plants. The expression levels were analyzed using reverse transcription–quantitative polymerase chain reaction (RT-qPCR) analysis. The qRT-PCR analysis of FPPS and GPPS gene expression was performed. The outputs of FPPS1 gene expression demonstrated high levels of mRNA in both S. alba and P. graveolens with fold changes of 25.24 and 21.68, respectively. In contrast, the minimum expression levels were observed for the GPPS1 gene, with fold changes of 11.28 and 6.48 in S. alba and P. graveolens, respectively. Thus, this study offers the employment of medicinal plants as an alternative to fertilizer usage resulting in promoting environmental preservation, optimal waste utilization, reducing water consumption, and cost reduction. Full article

This study investigates the impact of methyl jasmonate (MeJA) on the volatile oil composition of Schizonepeta tenuifolia and elucidates the function of the StTPS45 gene, a key player in terpenoid biosynthesis. The effect of different concentrations of MeJA (0, 50, 100, 200, and 300 μmol/L) on the growth of S. tenuifolia adventitious bud clusters was analyzed over a 20 d period. Using gas chromatography–mass spectrometry (GC-MS), 17 compounds were identified from the adventitious bud clusters of S. tenuifolia. Significant changes in the levels of major monoterpenes, including increased contents of (+)-limonene and (+)-menthone, were observed, particularly at higher concentrations of MeJA. Analysis of transcriptome data from three groups treated with 0, 100, and 300 μmol/L MeJA revealed significant changes in the gene expression profiles following MeJA treatment. At 100 μmol/L MeJA, most terpene synthase (TPS) genes were overexpressed. Additionally, gene expression and functional predictions suggested that StTPS45 acts as germacrene D synthase. Therefore, StTPS45 was cloned and expressed in Escherichia coli, and enzyme activity assays confirmed its function as a germacrene D synthase. Molecular docking and structural prediction of StTPS45 further suggested specific interactions with farnesyl diphosphate (FPP), aligning with its role in the terpenoid synthesis pathway. These findings provide valuable insights into the modulation of secondary metabolite pathways by jasmonate signaling and underscore the potential of genetic engineering approaches to enhance the production of specific terpenoids in medicinal plants. Full article

Aminobisphosphonates (NBPs) are the first-choice medication for osteoporosis (OP); NBP treatment aims at increasing bone mineral density (BMD) by inhibiting the activity of farnesyl diphosphate synthase (FDPS) enzyme in osteoclasts. Despite its efficacy, inadequate response to the drug and side effects have been reported. The A allele of the rs2297480 (A > C) SNP, found in the regulatory region of the FDPS gene, is associated with reduced gene transcription. This study evaluates the FDPS variant rs2297480 (A > C) association with OP patients’ response to alendronate sodium treatment. A total of 304 OP patients and 112 controls were enrolled; patients treated with alendronate sodium for two years were classified, according to BMD variations at specific regions (lumbar spine (L1-L4), femoral neck (FN) and total hip (TH), as responders (OP-R) (n = 20) and non-responders (OP-NR) (n = 40). We observed an association of CC genotype with treatment failure (p = 0.045), followed by a BMD decrease in the regions L1-L4 (CC = −2.21% ± 2.56; p = 0.026) and TH (CC = −2.06% ± 1.84; p = 0.015) after two years of alendronate sodium treatment. Relative expression of the FDPS gene was also evaluated in OP-R and OP-NR patients. Higher expression of the FDPS gene was also observed in OP-NR group (FC = 1.84 ± 0.77; p = 0.006) when compared to OP-R. In conclusion, the influence observed of FDPS expression and the rs2897480 variant on alendronate treatment highlights the importance of a genetic approach to improve the efficacy of treatment for primary osteoporosis. Full article

Farnesyl pyrophosphate synthase (FPPS) catalyzes the synthesis of C15 farnesyl diphosphate (FPP) from C5 dimethylallyl diphosphate (DMAPP) and two or three C5 isopentenyl diphosphates (IPPs). FPP is an important precursor for the synthesis of isoprenoids and is involved in multiple metabolic pathways. Here, farnesyl pyrophosphate synthase from Sporobolomyces pararoseus NGR (SpFPPS) was isolated and expressed by the prokaryotic expression system. The SpFPPS full-length genomic DNA and cDNA are 1566 bp and 1053 bp, respectively. This gene encodes a 350-amino acid protein with a predicted molecular mass of 40.33 kDa and a molecular weight of 58.03 kDa (40.33 kDa + 17.7 kDa), as detected by SDS-PAGE. The function of SpFPPS was identified by induction, purification, protein concentration and in vitro enzymatic activity experiments. Structural analysis showed that Y90 was essential for chain termination and changing the substrate scope. Site-directed mutation of Y90 to the smaller side-chain amino acids alanine (A) and lysine (K) showed in vitro that wt-SpFPPS catalyzed the condensation of the substrate DMAPP or geranyl diphosphate (GPP) with IPP at apparent saturation to synthesize FPP as the sole product and that the mutant protein SpFPPS-Y90A synthesized FPP and C20 geranylgeranyl diphosphate (GGPP), while SpFPPS-Y90K hydrolyzed the substrate GGPP. Our results showed that FPPS in S. pararoseus encodes the SpFPPS protein and that the amino acid substitution at Y90 changed the distribution of SpFPPS-catalyzed products. This provides a baseline for potentially regulating SpFPPS downstream products and improving the carotenoid biosynthesis pathway. Full article