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1.
Food Chem ; 400: 134016, 2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36084588

RESUMO

This work aims to investigate antifungal, mycotoxin inhibitory efficacy of the hop essential oil (HEO) nanoemulsion and their mode of action (MOA) against Fusarium graminearum isolate, a fungal pathogen causing Fusarium Head Blight (FHB) in cereal crops. The HEO, primarily consisting of terpenes and terpenoids, was encapsulated in nanoemulsion droplets. Physically stable HEO-in-water nanoemulsion was fabricated using 0.5 wt% of tween 80 and 5 wt% oil phase comprising 30 % of Ostwald ripening inhibitor and 70 % of HEO. In terms of antifungal effect, HEO nanoemulsion could not only effectively inhibit mycelial growth and spore germination of F. graminearum isolates, but also remarkably suppress the production of deoxynivalenol (DON) and its derivatives in rice culture by applying 750 µg of HEO/g rice. Our studies on the MOA showed that HEO nanoemulsion could alter the contents of total lipid and chitin in outer cell membrane as well as damaging cytoplasmic membrane.


Assuntos
Fusarium , Micotoxinas , Óleos Voláteis , Antifúngicos/metabolismo , Antifúngicos/farmacologia , Quitina/metabolismo , Fusarium/metabolismo , Micotoxinas/metabolismo , Óleos Voláteis/metabolismo , Óleos Voláteis/farmacologia , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Polissorbatos/farmacologia , Terpenos/metabolismo , Água/metabolismo
2.
J Ethnopharmacol ; 300: 115714, 2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36113678

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Swertia chirayita (Roxb.) H. Karst. is a traditionally used, well-recognized medicinal plant of the family Gentianaceae with significant therapeutic potential. It has been traditionally used to cure various ailments such as fever, vomiting, jaundice, digestive disorders, heart diseases, diabetes, malaria, scorpion bite, and skin diseases. AIM OF REVIEW: The present review emphasized the traditional uses, phytochemistry, pharmacology, toxicology, chemical profiling, and structural identification of isolated compounds by analytical and spectroscopic techniques. This review demonstrates the possibility of advanced ethnopharmacological research. MATERIALS AND METHODS: The literature on S. chirayita was obtained from bibliographic databases like Web of Science, PubMed, Science-Direct, American Chemical Society (ACS), Google Scholar, and SciFinder. The compiled review is covered up until March 2022. RESULTS: Approximately, 123 specialized metabolites including xanthones, seco-iridoids, terpenoids, alkaloids, and flavonoids have been isolated and characterized from S. chirayita. The extract and isolated compounds exhibited a wide spectrum of pharmacological effects such as anti-inflammatory, antioxidant, antitumor, hepatoprotective, antiviral, antimalarial, and antibacterial offering scientific evidence for traditional claims of this medicinal plant. In addition, various analytical methods using HPTLC, UPLC, HPLC, LC-MS, and GC-MS have also been documented to determine the phytochemicals of S. chirayita. CONCLUSION: The current article provides information on traditional usage, phytochemistry, chemical profiling, structure elucidation, pharmacological efficacy, toxicity, and future prospects of S. chirayita. This plant has long been traditionally used in a variety of ways by indigenous people. Numerous phytoconstituents and several pharmacological activities have been reported in S. chirayita. However, there are still some scientific gaps such as identification of bioactive compounds, structure-activity relationship and mechanistic action of isolated bioactive compounds, development of effective analytical methods for comprehensive quality control, and safety profiles that need to be addressed.


Assuntos
Antimaláricos , Plantas Medicinais , Swertia , Xantonas , Antibacterianos/uso terapêutico , Antimaláricos/uso terapêutico , Antioxidantes , Antivirais , Etnofarmacologia , Flavonoides , Humanos , Iridoides , Compostos Fitoquímicos/farmacologia , Compostos Fitoquímicos/uso terapêutico , Fitoterapia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Terpenos
3.
J Ethnopharmacol ; 300: 115748, 2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36162545

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: With high mortality and morbidity rates, lung cancer (LC) has become one of the major threats to human health. The treatment strategies for LC currently face issues, such as drug resistance and body tolerance. Traditional Chinese medicine (TCM) is characterized by novel pharmacological mechanisms, low toxicity, and limited side effects. TCM includes a substantial number of biologically active ingredients, several of which are effective monomeric agents against LC. An increasing number of researchers are focusing their efforts on the discovery of active anti-cancer ingredients in TCM. AIM OF THE REVIEW: In this review, we summarized the anti-LC mechanisms of five types of TCM monomeric compounds. Our goal is to provide research ideas for the identification of new prospective medication candidates for the treatment of LC. MATERIALS AND METHODS: We collected reports on the anti-LC effects of TCM monomers from web databases, including PubMed, Science Direct, Web of Science, and Europe PubMed Central. Among the keywords used were "lung cancer," "traditional Chinese medicine," "pharmacology," and their combinations thereof. Then, we systematically summarized the anti-LC efficacy and related mechanisms of TCM monomers. RESULTS: Based on the available literature, this paper reviewed the therapeutic effects and mechanisms of five types of TCM monomers on LC. The characteristics of TCM monomers include the capabilities to suppress the tumor cell cycle, inhibit proliferation, induce apoptosis, promote autophagy, inhibit tumor cell invasion and metastasis, and enhance efficacy or reduce drug resistance when combined with cytotoxic agents and other methods to arrest the progression of LC and prolong the survival of patients. CONCLUSIONS: TCM contains numerous flavonoids, alkaloids, terpenoids, polyphenols, and other active compounds that are effective against LC. Given their chemical structure and pharmacological properties, these monomers are suitable as candidate drugs for the treatment of LC.


Assuntos
Medicamentos de Ervas Chinesas , Neoplasias Pulmonares , Citotoxinas , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/uso terapêutico , Flavonoides , Humanos , Neoplasias Pulmonares/tratamento farmacológico , Medicina Tradicional Chinesa , Estudos Prospectivos , Terpenos
4.
Acta Trop ; 237: 106741, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36334844

RESUMO

BACKGROUND: Currently, symptomatic toxoplasmosis is treated with a combination of sulfadiazine and pyrimethamine. However, significant adverse effects and drug resistance have been reported. Terpenoids are widely found in nature, with numerous studies demonstrating that they have effective inhibitory effects on a variety of parasites. METHODS: In this study, we employed intermediates or derivatives of lindenane sesquiterpenoids to evaluate their intracellular and extracellular inhibitory effects on Toxoplasma gondii tachyzoites and their cytotoxicity on macrophages. RESULTS: We demonstrated that two of these target terpenoids could effectively reduce the number of extracellular tachyzoites, probably by inducing tachyzoite apoptosis through altering tachyzoites mitochondrial membrane potential and calcium homeostasis. In addition, the two target terpenoids were able to promote intracellular tachyzoites elimination, possibly by enhancing macrophage activities. CONCLUSIONS: Two derivatives of lindenane sesquiterpenoids with low cytotoxicity to macrophages demonstrated direct and indirect antitoxoplasma effects.


Assuntos
Parasitos , Toxoplasma , Toxoplasmose , Animais , Humanos , Terpenos/farmacologia , Toxoplasmose/tratamento farmacológico , Toxoplasmose/parasitologia , Pirimetamina/farmacologia
5.
Food Chem ; 403: 134421, 2023 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-36358073

RESUMO

The cluster-zone of the fan training system with multiple trunks (F-MT) distribute >1.5 m vertically and 1.0 m horizontally, which leads to microclimate heterogeneity around clusters. In the current study, clusters were divided into eight spatial positions according to the cluster growing height and light conditions, and the aroma profiles of grapes and wines were evaluated by HS-SPME-GC-MS. Results showed that the microclimate varied in different spatial positions. Light exposure promotes the accumulation of terpenes and C13-norisoprenoids, while inhibiting C6/C9 compounds in grape berries. Zone 2 wine presented the highest global aroma concentration. Floral and fruity were the main aromas in F-MT wines, but the herbaceous was more prominent in lower-position wines. C6/C9 compounds in grapes negatively correlated with terpenes and C13-norisoprenoids in wines. In conclusion, the aroma profiles of grapes and wines varied from different cluster positions, and graded harvesting will be helpful to produce quality wines.


Assuntos
Vitis , Compostos Orgânicos Voláteis , Vinho , Vinho/análise , Norisoprenoides , Microclima , Compostos Orgânicos Voláteis/análise , Odorantes/análise , Frutas/química , Terpenos
6.
Enzyme Microb Technol ; 162: 110148, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36343469

RESUMO

Terpenoids and steroids are two large families of natural products with diverse biological activities. C3 oxidase and acetyltransferase are the key enzymes in the post-modification of terpenoids and steroids, while their discovery is of great importance for the efficient bioproduction and wide application of these natural products. In this review, we first explain the reaction mechanism of typical enzymes. Next, we summarize the current state-of-art strategies in the discovery of C3 oxidase and acetyltransferase, and discuss rational engineering of these enzymes with novel catalytic functions. Finally, we propose how to obtain an excellent biocatalyst of C3 oxidase and acetyltransferase by exploiting the discovery strategy and the reaction mechanism.


Assuntos
Produtos Biológicos , Terpenos , Engenharia Metabólica , Oxirredutases , Acetiltransferases , Esteroides
7.
Food Chem ; 404(Pt B): 134726, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36327504

RESUMO

The rising interest in craft beer urging for novel yeasts that provide diverse attributes. This research aimed to explore the potential of four non-Saccharomyces yeasts with high ß-glucosidase activity in beer fermentation. Fermentation ability, adverse environment resistance and carbohydrate affinity were measured via growth and physiochemical parameters and, based on these results, a Candida glabrata (D18) yeast was selected for subsequent sequential fermentations. Aroma analysis of the fermented beer was then performed using gas chromatography-mass spectrometry, demonstrated that some glucoside-binding terpenes had been significantly increased. In two D18 involved groups, geraniol content increased 51.5/11-fold in the single and mixed fermentations, respectively, while linalool increased 1.36-fold in the mixed fermentation, providing the final beer product with unique floral and fruity characteristics. Principal component analysis and sensory evaluation were also applied to further demonstrate these distinctions. Overall, D18 was found to be a potential yeast for beer fermentation with unique properties.


Assuntos
Cerveja , Celulases , Cerveja/análise , Terpenos/análise , Leveduras , Fermentação
8.
Methods Mol Biol ; 2603: 127-138, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36370275

RESUMO

Chemical proteomics has been widely applied in the identification and quantification of targeted proteins. Here we describe a chemoproteomic method, in combination with stable isotope labeling by amino acids in cell culture (SILAC), for the proteome-wide profiling of geranyl pyrophosphate (GPP)-binding proteins. After labeling using a desthiobiotin-GPP acyl phosphate probe, desthiobiotin-conjugated peptides of GPP-binding proteins could be enriched from the tryptic digestion products of complex protein mixtures and subsequently identified with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. To exclude nonspecific binding proteins, we applied SILAC, together with competitive labeling experiments, including high vs. low concentrations of GPP probe, GPP vs. ATP probes, and GPP probe labeling with or without the presence of GPP. Several known or candidate GPP-binding proteins were identified with this method, suggesting the potential application of this method in the study of isoprenoid-interacting proteins and biological functions of isoprenoids.


Assuntos
Proteínas de Transporte , Espectrometria de Massas em Tandem , Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem/métodos , Proteômica/métodos , Proteínas , Terpenos , Marcação por Isótopo/métodos , Proteoma/análise
9.
Food Chem ; 404(Pt B): 134723, 2023 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-36444084

RESUMO

Essential oils (EOs) have recently gained popularity as natural food preservatives due to their potent antibacterial activity against food pathogens. In this review, the antibacterial activity of EOs from various plant parts and sources against the most important food pathogens Salmonella and Listeria have been discussed. The antibacterial activity of EOs is attributed to their major and minor low-molecular weight terpenes, terpenoids, phenylpropenes and aliphatic components. The major compounds along with minor components of EO extracted from different parts of various plant species were found to be responsible for antibacterial activity. The combination of EO from different sources presented synergistic anti-listerial and anti-salmonella effects. EO combined with biopolymer and in nanoemulsion form presented significant antibacterial activity. The mode of antibacterial action by EO was complex and involves a series of event that has also been discussed in detail.


Assuntos
Listeria , Óleos Voláteis , Óleos Voláteis/farmacologia , Salmonella , Antibacterianos/farmacologia , Terpenos
10.
J Ethnopharmacol ; 300: 115695, 2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36108894

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Veronicastrum Heist. ex Fabr. (Plantaginaceae) is a multifunctional plant in China and other parts of the continent. It has traditionally been used in the treatment of ascites, edema, blood stasis, pain relief, chronic nephritis injury, fever, cough, headache, arthritis, dysentery, rheumatism, pleural effusion, liver damage, and other disorders. Although research has confirmed that the genus Veronicastrum contain many active compounds, no review of its traditional uses, phytochemistry or pharmacology has been conducted to date. AIM: This review aims to systematically evaluate the traditional uses, phytochemistry, and pharmacology of the genus Veronicastrum, discuss its medicinal potential, modern scientific research, and the relationship between them, and put forward some suggestions to promote further development and utilization of Veronicastrum. MATERIALS AND METHODS: The traditional uses, phytochemical and pharmacological data related to the genus Veronicastrum from 1955 to date was compiled by surveying the ethnomedicinal books and published papers, and searching the online databases including Google Scholar, China National Knowledge Infrastructure (CNKI), Science Direct, Web of Science and World Flora Online. RESULTS: Species of the genus Veronicastrum are widely used in folkloric medicine and some of their uses have been confirmed in modern pharmacological activities. A total of 89 chemical constituents have been isolated from the genus Veronicastrum, including flavonoids, carbohydrates, iridoids, terpenoids, phytosterols, phenolic acids, and other constituents. Among the compounds isolated, iridoids, flavonoids, and terpenoids are responsible for the biological activities of this genus with significant pharmacological activities both in vitro and in vivo. The extracts and compounds isolated from this genus have been reported to contain a wide range of pharmacological activities such as immunosuppressive, antioxidant, anti-cancer, anti-inflammatory, gastro protective, and antimicrobial activity. CONCLUSION: The genus Veronicastrum is not only a great herbal remedy, but also has numerous bioactive chemicals with potential for new drug discovery. In the literature, phytochemical investigations have been undertaken on five species. Detailed scientific research is still needed to fully understand this genus. Furthermore, its bioactive chemicals' structure-activity connection, in vivo activity, and mechanism of action ought to be investigated further.


Assuntos
Anti-Infecciosos , Fitosteróis , Plantaginaceae , Antioxidantes , Carboidratos , Etnofarmacologia , Flavonoides , Iridoides , Compostos Fitoquímicos/química , Compostos Fitoquímicos/farmacologia , Compostos Fitoquímicos/uso terapêutico , Fitoterapia , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Extratos Vegetais/uso terapêutico , Terpenos
11.
Food Chem ; 402: 134209, 2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36126576

RESUMO

Cocoa powder is a highly consumed product all over the world which could be substituted by cheaper raw materials resulting in food fraud. In this work, a non-targeted metabolomics approach based on the use of reversed-phase liquid chromatography coupled to high-resolution mass spectrometry was developed to carry out the characterization of cocoa powder samples adulterated, at two different levels, with carob flour, soy flour, and chicory. The sample preparation protocol and the chromatographic parameters were optimized to extract and detect the highest number of molecular features. Both non-supervised and supervised statistical methods were employed to analyze the most significant variables that gave rise to group discrimination. From the 21 and 37 significant variables analyzed in positive and negative ionization modes, respectively, a total of 20 were tentatively identified. Different families of compounds including flavonoids, fatty acids, terpenoids, lysophospholipids, and a galactolipid could be pointed out as cocoa adulteration markers.


Assuntos
Chocolate , Cromatografia Líquida de Alta Pressão/métodos , Chocolate/análise , Galactolipídeos , Espectrometria de Massas/métodos , Metabolômica/métodos , Flavonoides/análise , Ácidos Graxos , Lisofosfolipídeos , Terpenos/análise
12.
J. Health Biol. Sci. (Online) ; 10(1): 1-10, 01/jan./2022. tab, ilus
Artigo em Inglês | LILACS | ID: biblio-1378456

RESUMO

Objective: This study aimed to evaluate the interactions of di- and tri-terpenes from Stillingia loranthacea with the enzyme NSP16-NSP10 of SARS-CoV-2, important for viral replication. Methods: The molecular docking technique was used to evaluate this interaction. Results: The analysis showed that the evaluated compounds obtained RMSD values of 0.888 to 1.944 Å and free energy of -6.1 to -9.4 kcal/mol, with the observation of hydrogen bonds, salt bridges, and pi-sulfur, pi-alkyl, and hydrophobic interactions. Conclusion: Thus, the results obtained show the potential of the compounds analyzed against the selected target. Since computer simulations are only an initial step in projects for the development of antiviral drugs, this study provides important data for future research.


Objetivo: avaliar as interações de di- e tri-terpenos de Stillingia loranthacea com a enzima NSP16-NSP10 de SARS-CoV-2, importante para a replicação viral. Métodos: A técnica de docking molecular foi utilizada para avaliar essa interação. Resultados: A análise mostrou que os compostos avaliados obtiveram valores de RMSD de 0,888 a 1,944 Å e energia livre de -6,1 a -9,4 kcal/mol, observando-se ligações de hidrogênio, pontes salinas e pi-enxofre, pi-alquil, e interações hidrofóbicas. Conclusão: Assim, os resultados obtidos mostram o potencial dos compostos analisados frente ao alvo selecionado. Como as simulações computacionais são apenas um passo inicial nos projetos de desenvolvimento de medicamentos antivirais, este estudo fornece dados importantes para pesquisas futuras.


Assuntos
SARS-CoV-2 , Antivirais , Terpenos , Replicação Viral , Enzimas , Simulação de Acoplamento Molecular
13.
Nat Prod Res ; 36(22): 5753-5761, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36411528

RESUMO

Two new terpenoids (1 and 2), arenterpenoid D (1) and pinnasesquiterpene A (2), along with 16 phenylpropanoids (3-18) and 8 known terpenoids (19-26) were isolated from 70% EtOH extract of the Arenga pinnata (Wurmb) Merr. fruits. Their structures were elucidated by spectroscopic methods including HR-ESI-MS, 1D, and 2D NMR. The absolute configuration of arenterpenoid D (1) was defined by X-ray crystallographic analysis. Furthermore, we evaluated the anti-inflammatory activity of all compounds, and outcomes showed that 2 and 21 exposed moderate suppressive effects against NO generation in lipopolysaccharide-stimulated RAW 264.7 cells.


Assuntos
Arecaceae , Terpenos , Camundongos , Animais , Terpenos/farmacologia , Terpenos/análise , Frutas/química , Arecaceae/química , Anti-Inflamatórios/química , Células RAW 264.7
14.
Ecotoxicol Environ Saf ; 247: 114261, 2022 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-36332404

RESUMO

The entry mode of terpenes into the atmosphere is via volatilization of hydrocarbons from foliage over heavily forested areas besides entering the environment through surface water runoff. Some monoterpenes in essential oils are phytotoxins, acting as plant chemical defenses against bacteria or fungi infections and plant-eating insects. For organisms to survive, their enzymatic systems are activated in response to an assault by potentially harmful compounds. Certain bacterial and fungal genera have developed special abilities to transform toxic terpenes into less toxic derivatives. Here, we investigated the response of the bacterial and fungal community in Haemaphysalis longicornis exposed to Cymbopogon citratus (lemongrass) essential oil (EO) and citronellal. Sequencing of bacterial 16S rRNA and fungal ITS1 regions on an Illumina NovaSeq PE250 sequencing platform was performed for H. longicornis tick samples treated with 15 and 20 mg/mL of lemongrass essential oil and citronellal. The diversity recorded in samples treated with C. citratus EO was higher in comparison to those treated with citronellal but significantly lower in the control samples as reflected by the Shannon diversity index. All major H. longicornis bacterial phyla, including Proteobacteria (93.81 %), Firmicutes (2.58 %), and Bacteroidota (0.99 %) were detected. A switch of dominance from Coxiella to Pseudomonas, which has high biotransformation capacity, was observed in the bacterial community, whereas the phylum Ascomycota (Genera: Aspergillus, Archaeorhizomyces, Alternaria, and Candida) dominated in the fungal community indicating detoxifying symbiosis. Other significantly abundant bacterial genera include Ralstonia, Acinetobacter, Vibrio, and Pseudoalteromonas, while Ganoderma and Trichosporon (yeasts) spp. represented the fungi Basidiomycota. This study expanded the understanding of enzymatic modification of phytotoxic substances by microorganisms, which could provide deeper insights into the mitigation of harmful phytotoxins and the synthesis of eco-friendly derivatives for the control of ticks.


Assuntos
Cymbopogon , Óleos Voláteis , Carrapatos , Animais , Simbiose , Monoterpenos/farmacologia , RNA Ribossômico 16S/genética , Bactérias/genética , Óleos Voláteis/farmacologia , Terpenos , Fungos/genética
15.
Plant Signal Behav ; 17(1): 2138041, 2022 12 31.
Artigo em Inglês | MEDLINE | ID: mdl-36317599

RESUMO

Ulmus parvifolia (U. parvifolia) is a Chinese medicine plant whose bark and leaves are used in the treatment of some diseases such as inflammation, diarrhea and fever. However, metabolic signatures of seeds have not been studied. The seeds and bark of U. parvifolia collected at the seed ripening stage were used for metabolite profiling analysis through the untargeted metabolomics approach. A total of 2,578 and 2,207 metabolites, while 503 and 132 unique metabolites were identified in seeds and bark, respectively. Additionally, 574 differential metabolites (DEMs) were detected in the two different organs of U. parvifolia, which were grouped into 52 classes. Most kinds of metabolites classed into prenol lipids class. The relative content of flavonoids class was the highest. DEMs contained some bioactive compounds (e.g., flavonoids, terpene glycosides, triterpenoids, sesquiterpenoids) with antioxidant, anti-inflammatory, and anti-cancer activities. Most kinds of flavonoids and sesquiterpenes were up-regulated in seeds. There were more varieties of terpene glycosides and triterpenoids showing up-regulated in bark. The pathway enrichment was performed, while flavonoid biosynthesis, flavone and flavonol biosynthesis were worthy of attention. This study identified DEMs with pharmaceutical value between seeds and bark during seed maturation and offered a molecular basis for alternative or complementary use of seeds and bark of U. parvifolia as a Chinese medicinal material.


Assuntos
Triterpenos , Ulmus , Ulmus/metabolismo , Casca de Planta/metabolismo , Medicina Tradicional Chinesa , Extratos Vegetais , Sementes/metabolismo , Flavonoides/metabolismo , Glicosídeos/metabolismo , Triterpenos/metabolismo , Terpenos/metabolismo
16.
Int J Mol Sci ; 23(21)2022 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-36361694

RESUMO

Dimethylallyl diphosphate (DMAPP) is a key intermediate metabolite in the synthesis of isoprenoids and is also the prenyl donor for biosynthesizing prenylated flavonoids. However, it is difficult to prepare DMAPP via chemical and enzymatic methods. In this study, three promiscuous kinases from Shigella flexneri (SfPK), Escherichia coli (EcPK), and Saccharomyces cerevisiae (ScPK) and three isopentenyl phosphate kinases from Methanolobus tindarius (MtIPK), Methanothermobacter thermautotrophicus str. Delta H (MthIPK), and Arabidopsis thaliana (AtIPK) were cloned and expressed in Escherichia coli. The enzymatic properties of recombinant enzymes were determined. The Kcat/Km value of SfPK for DMA was 6875 s-1 M-1, which was significantly higher than those of EcPK and ScPK. The Kcat/Km value of MtIPK for DMAP was 402.9 s-1 M-1, which was ~400% of that of MthIPK. SfPK was stable at pH 7.0-9.5 and had a 1 h half-life at 65 °C. MtIPK was stable at pH 6.0-8.5 and had a 1 h half-life at 50 °C. The stability of SfPK and MtIPK was better than that of the other enzymes. Thus, SfPK and MtIPK were chosen to develop a one-pot enzymatic cascade for producing DMAPP from DMA because of their catalytic efficiency and stability. The optimal ratio between SfPK and MtIPK was 1:8. The optimal pH and temperature for the one-pot enzymatic cascade were 7.0 and 35 °C, respectively. The optimal concentrations of ATP and DMA were 10 and 80 mM, respectively. Finally, maximum DMAPP production reached 1.23 mM at 1 h under optimal conditions. Therefore, the enzymatic method described herein for the biosynthesis of DMAPP from DMA can be widely used for the synthesis of isoprenoids and prenylated flavonoids.


Assuntos
Hemiterpenos , Fosfatos , Fosfatos/metabolismo , Escherichia coli/metabolismo , Organofosfatos/metabolismo , Terpenos/metabolismo , Flavonoides/metabolismo
17.
Appl Microbiol Biotechnol ; 106(23): 7779-7791, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36350403

RESUMO

Ganoderma sinense, with more than 2000 years of medicinal history, is a fungus of the basidiomycetes that is rich in polysaccharides and terpenoids. However, the biosynthesis of terpenes, especially sesquiterpenes, has been little studied. The functional identification of sesquiterpene synthases from G. sinense is of great significance to the study of fungal terpenoid biosynthesis and regulation. Our research group has completed the functional characterization of 21 sesquiterpene synthase genes from G. sinense. It was found that gleenol, biosynthesis of which is catalyzed by the sesquiterpene synthase GsSTS26 and GsSTS27, has the functions of killing termites, antihelminth, and plant growth regulation. In the unmodified E. coli Rosetta (DE3) strain, the content of gleenol produced by sesquiterpene synthase from G. sinense is low, which makes it difficult to meet the demand of industrial production and the market. Therefore, it is of great significance to obtain high-yielding strains by means of synthetic biology. In this study, we constructed eight recombinant strains by using tandem gene expression and promoter engineering, and the content of gleenol was increased by up to 23-fold. In this study, we realized the de novo synthesis of gleenol in E. coli and provided a basis for the biosynthesis of terpenoids in basidiomycetes. KEY POINTS: • Eight recombinant expression systems were constructed by using tandem genes and promoter engineering. • The recombinant strain promoted the efficient production of gleenol in E. coli Rosetta (DE3). • The recombinant strain achieved de novo production of gleenol in E. coli.


Assuntos
Alquil e Aril Transferases , Sesquiterpenos , Escherichia coli/genética , Escherichia coli/metabolismo , Sesquiterpenos/metabolismo , Terpenos/metabolismo , Alquil e Aril Transferases/metabolismo
18.
Metab Eng ; 74: 139-149, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36341776

RESUMO

The production of terpenoids from engineered microbes contributes markedly to the bioeconomy by providing essential medicines, sustainable materials, and renewable fuels. The mevalonate pathway leading to the synthesis of terpenoid precursors has been extensively targeted for engineering. Nevertheless, the importance of individual pathway enzymes to the overall pathway flux and final terpenoid yield is less known, especially enzymes that are thought to be non-rate-limiting. To investigate the individual contribution of the five non-rate-limiting enzymes in the mevalonate pathway, we created a combinatorial library of 243 Saccharomyces cerevisiae strains, each having an extra copy of the mevalonate pathway integrated into the genome and expressing the non-rate-limiting enzymes from a unique combination of promoters. High-throughput screening combined with machine learning algorithms revealed that the mevalonate kinase, Erg12p, stands out as the critical enzyme that influences product titer. ERG12 is ideally expressed from a medium-strength promoter which is the 'sweet spot' resulting in high product yield. Additionally, a platform strain was created by targeting the mevalonate pathway to both the cytosol and peroxisomes. The dual localization synergistically increased terpenoid production and implied that some mevalonate pathway intermediates, such as mevalonate, isopentyl pyrophosphate (IPP), and dimethylallyl pyrophosphate (DMAPP), are diffusible across peroxisome membranes. The platform strain resulted in 94-fold, 60-fold, and 35-fold improved titer of monoterpene geraniol, sesquiterpene α-humulene, and triterpene squalene, respectively. The terpenoid platform strain will serve as a chassis for producing any terpenoids and terpene derivatives.


Assuntos
Ácido Mevalônico , Saccharomyces cerevisiae , Ácido Mevalônico/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Terpenos/metabolismo , Difosfatos/metabolismo , Engenharia Metabólica/métodos , Aprendizado de Máquina
19.
Int J Mol Sci ; 23(22)2022 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-36430522

RESUMO

Phoebe hui is an extremely valuable tree that is the main source of the fragrant golden-thread nanmu wood. Although the fragrance of wood has been investigated in several trees, the potential substances and gene regulation mechanisms that are involved in fragrance formation are poorly understood. Here, three radial tissues, sapwood (SW), heartwood (HW), and the transition zone (TZ) in between them, were compared via integrative physiological, volatile-metabolomic, and transcriptomic analyses to identify the key metabolites and regulatory mechanisms involved in fragrance formation. During heartwood formation, gradual starch grain loss was accompanied by the deposition of lipids and extractives in the cell lumen. Extracts of terpenoids were synthesized and accumulated in the heartwood, including monoterpenoids (limonene and p-cymene) and sesquiterpenes (cubebene and guaiadiene); these were identified as being closely related to the special fragrance of the wood. Additionally, the expression of transcripts showed that the genes related to primary metabolism were specifically upregulated in the SW, whereas genes annotated in terpenoid biosynthesis were specifically upregulated in the HW. Therefore, we speculated that terpenoid biosynthesis occurs in situ in the HW via the HW formation model of Type-III (Santalum) using the precursors that were produced by primary metabolism in the SW. The expression levels of transcription factors (e.g., MYB, WRKY, and C2H2) acted as the major regulatory factors in the synthesis of terpenoids. Our results explain the special fragrance in P. hui and broaden the current knowledge of the regulatory mechanisms of fragrance formation. This work provides a framework for future research that is focused on improving wood quality and value.


Assuntos
Lauraceae , Perfumes , Santalum , Transcriptoma , Odorantes , Metabolômica , Santalum/genética , Perfumes/metabolismo , Lauraceae/genética , Terpenos/metabolismo , Árvores/genética
20.
Int J Mol Sci ; 23(22)2022 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-36430556

RESUMO

In this experimental research, different types of essential oils (EOs) were blended with polyhydroxybutyrate (PHB) to study the influence of these additives on PHB degradation. The blends were developed by incorporating three terpenoids at two concentrations (1 and 3%). The mineralization rate obtained from CO2 released from each sample was the factor that defined biodegradation. Furthermore, scanning electron microscope (SEM), differential scanning calorimetry (DSC), and dynamic mechanical analysis (DMA) were used in this research. The biodegradation percentages of PHB blended with 3% of eucalyptol, limonene, and thymol after 226 days were reached 66.4%, 73.3%, and 76.9%, respectively, while the rate for pure PHB was 100% after 198 days, and SEM images proved these results. Mechanical analysis of the samples showed that eucalyptol had the highest resistance level, even before the burial test. The other additives showed excellent mechanical properties although they had less mechanical strength than pure PHB after extrusion. The samples' mechanical properties improved due to their crystallinity and decreased glass transition temperature (Tg). DSC results showed that blending terpenoids caused a reduction in Tg, which is evident in the DMA results, and a negligible reduction in melting point (Tm).


Assuntos
Anti-Infecciosos , Butiratos , Poliésteres/química , Terpenos , Eucaliptol , Antibacterianos
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