Production Of Phenolic Acids Research Articles

Ekstraksi Daun Kejibeling (Strobilanthes crispus) dengan Metode Continuous Ultrasound-Assisted Enzymatic Extraction: Pengaruh Temperatur dan Konsentrasi Enzim

Continuous ultrasound-assisted enzymatic extraction (CUAEE) method has become a promising technique for extracting bioactive compounds from plant materials. In this study, the extraction of kejibeling (Strobilanthes crispus) leaves was carried out using the CUAEE method to optimize the extract yield. The experimental design involved extraction parameters including leaf particle size (d ≤0,177 mm), ethanol concentration (50%), extraction time (180 minutes), and varying parameters such as temperature (30 °C, 40 °C, 50 °C, and 60 °C), as well as cellulase enzyme concentration of 3% and 5%. The results showed that the optimal conditions for CUAEE were achieved at a temperature of 60 °C with an enzyme-solid ratio of 5%, producing a total phenolic content (TPC) of 12,097 mg EAG/g dry leaf powder and a total flavonoid content (TFC) of 2,291 EK/g dry leaf powder. The data obtained indicate that the continuous ultrasound-assisted enzymatic extraction method has the potential for large-scale production of phenolic acids and flavonoids.

Metabolic engineering of artificially modified transcription factor SmMYB36-VP16 for high-level production of tanshinones and phenolic acids

Tanshinones and phenolic acids are the two main chemical constituents in Salvia miltiorrhiza, which are used clinically for the treatment of hypertension, coronary heart disease, atherosclerosis, and many other diseases, and have broad medicinal value. The efficient synthesis of the target products of these two metabolites in isolated plant tissues cannot be achieved without the regulation and optimization of metabolic pathways, and transcription factors play an important role as common regulatory elements in plant tissue metabolic engineering. However, most of the regulatory effects are specific to one class of metabolites, or an opposing regulation of two classes of metabolites exists. In this study, an artificially modified transcription factor, SmMYB36-VP16, was constructed to enhance tanshinones and phenolic acids in Salvia miltiorrhiza hair roots simultaneously. Further in combination with the elicitors dual-screening technique, by applying the optimal elicitors screened, the tanshinones content in the transgenic hairy roots of Salvia miltiorrhiza reached 6.44 mg/g DW, which was theoretically 6.08-fold that of the controls without any treatment, and the content of phenolic acids reached 141.03 mg/g DW, which was theoretically 5.05-fold that of the controls without any treatment. The combination of artificially modified transcriptional regulatory and elicitors dual-screening techniques has facilitated the ability of plant isolated tissue cell factories to produce targeted medicinal metabolites. This strategy could be applied to other species, laying the foundation for the production of potential natural products for the medicinal industry.

Ag+ ions are effective elicitors for enhancing the production of phenolic acids and tanshinones in Salvia aristata Aucher ex Benth. hairy roots

Ag+ ions are effective elicitors for enhancing the production of phenolic acids and tanshinones in Salvia aristata Aucher ex Benth. hairy roots

AMF inhibit the production of phenolic acid autotoxins at the seed-filling stage in soybeans with continuous monocropping

BackgroundSoybean is the main oil crop in Northeast China. Continuous monocropping is more commonly used for soybean production due to rising market demand and arable land constraints. However, autotoxic substances, such as phenolic acids, produced by continuously cropped soybean can reduce yield and quality. The mycorrhiza formed of Arbuscular mycorrhizal fungi (AMF) and plant roots regulate the metabolic activities of the host plant and increase its disease resistance. The main purpose of this study was to inhibit the production of phenolic acids and determine the adverse effects on the growth of continuous monocropping soybean by inoculating Funneliformis mosseae (F. mosseae).ResultsTranscriptomics results showed that the production of phenolic acids in continuous monocropping soybean roots was mainly regulated by the expression of the CHS6, PCL1, SAMT, SRG1, and ACO1 genes, and the expression of these genes was significantly downregulated after inoculation with F. mosseae. Metabolomics results showed that continuous monocropping soybean roots inoculated with F. mosseae inhibited phenolic acid production through the phenylpropane biosynthetic, α-linoleic acid, linoleic acid, and other metabolic pathways. Phenolic acids in the phenylpropane metabolic pathway, such as 4-hydroxybenzoic acid, phthalic acid, and vanillic acid, decreased significantly after inoculation with F. mosseae. The combined analysis of the two showed that genes such as YLS9 and ARF3 were positively correlated with 4-hydroxybenzoic acid and so on, while genes such as CHS6 and SRG1 were negatively correlated with butyric acid and so on.ConclusionF. mosseae regulated the expression of functional genes and related phenolic acid metabolic pathways produced by continuous monocropping soybean roots, inhibiting the production of phenolic acid autotoxic substances in continuous cropped soybean, and slowing down the disturbance of continuous monocropping. This study provides a new solution for continuous monocropping of plants to overcome the autotoxicity barrier and provides a new basis for the development and utilization of AMF as a biological agent.

Withania somnifera (L.) Dunal: Enhanced production of withanolides and phenolic acids from hairy root culture after application of elicitors

Withania somnifera (L.) Dunal is an important indigenous medicinal plant with extensive pharmaceutical potential. The root is the main source of major bioactive compounds of this plant species including withanolides, withanine, phenolic acids, etc. Hairy root culture (HRC) is a crucial method for low-cost production of active compounds on a large scale. Four different Agrobacterium rhizogenes strains have been used for the hairy root induction. Maximum transformation efficiency (87.34 ± 2.13%) was achieved with A4 bacterial strain-mediated transformed culture. The genetic transformation was confirmed by using specific primers of seven different genes. Seven HR (Hairy root) lines were selected after screening 29 HR lines based on their fast growth rate and high accumulation of withanolides and phenolic acids content. Two biotic and three abiotic elicitors were applied to the elite root line to trigger more accumulation of withanolides and phenolic acids. While all the elicitors effectively increased withanolides and phenolic acids production, among the five different elicitors, salicylic acid (4.14 mg l–1) induced 11.49 -fold increase in withanolides (89.07 ± 2.75 mg g–1 DW) and 5.34- fold increase in phenolic acids (83.69 ± 3.11 mg g− 1 DW) after 5 days of elicitation compared to the non-elicited culture (7.75 ± 0.63 mg g–1 DW of withanolides and 15.66 ± 0.92 mg g–1 DW of phenolic acids). These results suggest that elicitors can tremendously increase the biosynthesis of active compounds in this system; thus, the HRC of W. somnifera is cost-effective and can be efficiently used for the industrial production of withanolides and phenolic acids.

Recent Advances in Microbial Metabolic Engineering for Production of Natural Phenolic Acids.

Phenolic acids are important natural bioactive compounds with varied physiological functions. They are extensively used in food, pharmaceutical, cosmetic, and other chemical industries and have attractive market prospects. Compared to plant extraction and chemical synthesis, microbial fermentation for phenolic acid production from renewable carbon sources has significant advantages. This review focuses on the structural information, physiological functions, current applications, and biosynthesis pathways of phenolic acids, especially advances in the development of metabolically engineered microbes for the production of phenolic acids. This review provides useful insights concerning phenolic acid production through metabolic engineering of microbial cell factories.

Shining a Light on Symbiosis: N-Fixing Bacteria Boost Legume Growth under Varied Light Conditions

Legumes are a diverse and important group of plants that play a vital role in agriculture, food security, and environmental sustainability. Rhizobia are symbiotic bacteria that form nitrogen-fixing nodules on legume roots, providing the plant with a valuable source of nitrogen. Phenolic acids are a group of secondary metabolites produced by plants that have a wide range of biological functions, including defense against pests and diseases, tolerance to abiotic stresses, and nutrient uptake. In the context of climate change and the imperative for sustainable agriculture, this study delves into the dynamic responses of legume species to varying light intensities and their intricate interactions with soil microorganisms. We investigated the impact of light intensity and rhizobial inoculation on the biomass, nitrate reductase, acid phosphatase, and production of phenolic acids in the roots of four legume species, Trifolium repens, Vicia sativa, Ornithopus compressus, and Coronilla juncea. Plants were grown under three light intensity regimes (low, medium, and high) and inoculated with either rhizobia or a non-inoculated control. The results highlight that shaded-light-adapted species, T. repens and V. sativa, increased root exudate production when exposed to high light intensity. This response aligns with their mining strategy, effectively allocating resources to optimize nutrient acquisition under varying conditions. In contrast, species hailing from well-illuminated environments, O. compressus and C. juncea, displayed distinct strategies by significantly increasing biomass under high irradiance, capitalizing on the available light and nutrients. The mining strategy of legumes emerged as a central theme, influencing biomass production, nitrogen dynamics, and enzymatic activities. The strong correlations between biomass and total nitrogen accumulation underscore the role of the mining strategy in efficient nutrient acquisition. Inoculated plants, which rely more on biological nitrogen fixation (BNF), exhibited lower δ15N values, indicative of a successful mining strategy to acquire and utilize atmospheric nitrogen. Enzymatic activities and phenolic acids exhibited significant interspecies variations, reflecting the adaptability of legumes to different light conditions. The findings of this study could be used to develop new strategies for improving legume stress tolerance, nutrient uptake capacity, and rhizosphere health.

Integrative effects of phytohormones in the phenolic acids production in Salvia verticillata L. under multi-walled carbon nanotubes and methyl jasmonate elicitation

Salvia verticillata L. is a well-known herb rich in rosmarinic acid (RA) and with therapeutic values. To better understand the possible roles of phytohormones in the production of phenolic acids in S. verticillata, in this work, we investigated some physiological and biochemical responses of the species to methyl jasmonate (MJ) and multi-walled carbon nanotubes (MWCNTs) as two effective elicitors. The leaves were sprayed with aqueous solutions containing 100 mg L−1 MWCNTs and 100 µM MJ and then harvested during interval times of exposure up to 96 h. The level of abscisic acid, as the first effective phytohormone, was altered in the leaves in response to MJ and MWCNTs elicitation (2.26- and 3.06-fold more than the control, respectively), followed by significant increases (P ˂ 0.05) detected in jasmonic acid and salicylic acid contents up to 8 h after exposure. Obtained data revealed that simultaneously with changes in phytohormone profiles, significant (P ˂ 0.05) rises were observed in the content of H2O2 (8.85- and 9.74-folds of control), and the amount of lipid peroxidation (10.18- and 17.01-folds of control) during the initial times after exposure to MJ and MWCNTs, respectively. Later, the content of phenolic acids increased in the elicited leaves due to changes in the transcription levels of key enzymes involved in their biosynthesis pathways, so 2.71- and 11.52-fold enhances observed in the RA content of the leaves after exposure to MJ and MWCNTs, respectively. It is reasonable to conclude that putative linkages between changes in some phytohormone pools lead to the accumulation of phenolic acids in the leaves of S. verticillata under elicitation. Overall, the current findings help us improve our understanding of the signal transduction pathways of the applied stimuli that led to enhanced secondary metabolite production in medicinal plants.

Abscisic Acid Affects Phenolic Acid Content to Increase Tolerance to UV-B Stress in Rhododendron chrysanthum Pall.

The presence of the ozone hole increases the amount of UV radiation reaching a plant's surface, and UV-B radiation is an abiotic stress capable of affecting plant growth. Rhododendron chrysanthum Pall. (R. chrysanthum) grows in alpine regions, where strong UV-B radiation is present, and has been able to adapt to strong UV-B radiation over a long period of evolution. We investigated the response of R. chrysanthum leaves to UV-B radiation using widely targeted metabolomics and transcriptomics. Although phytohormones have been studied for many years in plant growth and development and adaptation to environmental stresses, this paper is innovative in terms of the species studied and the methods used. Using unique species and the latest research methods, this paper was able to add information to this topic for the species R. chrysanthum. We treated R. chrysanthum grown in a simulated alpine environment, with group M receiving no UV-B radiation and groups N and Q (externally applied abscisic acid treatment) receiving UV-B radiation for 2 days (8 h per day). The results of the MN group showed significant changes in phenolic acid accumulation and differential expression of genes related to phenolic acid synthesis in leaves of R. chrysanthum after UV-B radiation. We combined transcriptomics and metabolomics data to map the metabolic regulatory network of phenolic acids under UV-B stress in order to investigate the response of such secondary metabolites to stress. L-phenylalanine, L-tyrosine and phenylpyruvic acid contents in R. chrysanthum were significantly increased after UV-B radiation. Simultaneously, the levels of 3-hydroxyphenylacetic acid, 2-phenylethanol, anthranilate, 2-hydroxycinnamic acid, 3-hydroxycinnamic acid, α-hydroxycinnamic acid and 2-hydroxy-3-phenylpropanoic acid in this pathway were elevated in response to UV-B stress. In contrast, the study in the NQ group found that externally applied abscisic acid (ABA) in R. chrysanthum had greater tolerance to UV-B radiation, and phenolic acid accumulation under the influence of ABA also showed greater differences. The contents of 2-phenylethanol, 1-o-p-coumaroyl-β-d-glucose, 2-hydroxy-3-phenylpropanoic acid, 3-(4-hydroxyphenyl)-propionic acid and 3-o-feruloylquinic ac-id-o-glucoside were significantly elevated in R. chrysanthum after external application of ABA to protect against UV-B stress. Taken together, these studies of the three groups indicated that ABA can influence phenolic acid production to promote the response of R. chrysanthum to UV-B stress, which provided a theoretical reference for the study of its complex molecular regulatory mechanism.

Phenolic Acids and Flavonoids Play Important Roles in Flower Bud Differentiation in Mikania micrantha: Transcriptomics and Metabolomics

Mikania micrantha is a highly invasive vine, and its ability to sexually reproduce is a major obstacle to its eradication. The long-distance dissemination of M. micrantha depends on the distribution of seeds; therefore, inhibiting M. micrantha flowering and seed production is an effective control strategy. The number of blooms of M. micrantha differs at different altitudes (200, 900, and 1300 m). In this study, we used a combination of metabolomics and transcriptomics methods to study the patterns of metabolite accumulation in the flower buds of M. micrantha. Using LC-MS/MS, 658 metabolites were found in the flower buds of M. micrantha at three different altitudes (200, 900, and 1300 m). Flavonoids and phenolic acids were found to be the main differential metabolites, and their concentrations were lower at 900 m than at 200 m and 1300 m, with the concentrations of benzoic acid, ferulic acid, and caffeic acid being the lowest. The biosynthesis pathways for flavonoids and phenolic compounds were significantly enriched for differentially expressed genes (DEGs), according to the results of transcriptome analysis. The production of flavonoid and phenolic acids was strongly linked with the expressions of phenylalanine ammonia-lyase (PAL), caffeoyl-CoA O-methyltransferase (COMT), and 4-coumarate-CoA ligase (4CL), according to the results of the combined transcriptome and metabolome analysis. These genes’ roles in the regulation of distinct phenolic acids and flavonoids during M. micrantha bud differentiation are still unknown. This study adds to our understanding of how phenolic acids and flavonoids are regulated in M. micrantha flower buds at various altitudes and identifies regulatory networks that may be involved in this phenomenon, offering a new approach for the prevention and management of M. micrantha.

Biochemical changes and enhanced accumulation of phenolic compounds in cell culture of Perilla frutescens (L.) by nano-chemical elicitation

Perilla frutescens (L.) Britt is a renowned medicinal plant with pharmaceutically valuable phenolic acids and flavonoids. The present study was aimed to study the eliciting effect of silver and copper nanoparticles (AgNPs and CuNPs, 50 and 100 mg/L), and methyl jasmonate (MeJa, 50 and 100 μM) on the biochemical traits, the accumulation of phenolic compounds and antioxidative capacity of P. frutescens cell suspension culture. Suspension cells were obtained from friable calli derived from nodal explants in Murashige and Skoog (MS) liquid medium containing 1 mg/L 2,4-D and 1 mg/L BAP. The 21 days old cell suspension culture established from nodal explant derived callus supplemented with 100 mg/L MeJa resulted in the highest activity of catalase and guaiacol peroxidase enzymes, and CuNPs 100 mg/L treated cells indicated the maximum content of total phenol, total anthocyanin, superoxide dismutase, malondialdehyde, and H2O2. Also, the highest content of ferulic acid (1.41 ± 0.03, mg/g DW), rosmarinic acid (19.29 ± 0.12, mg/g DW), and phenylalanine ammonia-lyase (16.81 ± 0.18, U/mg protein) were observed with 100 mg/L CuNPs, exhibiting a total increase of 1.58-fold, 2.12-fold, and 1.51-fold, respectively, higher than untreated cells. On the other hand, AgNPs 100 mg/L treated cells indicated the most amounts of caffeic acid (0.57 ± 0.03, mg/g DW) and rutin (1.13 ± 0.07, mg/g DW), as well as the highest scavenging potential of free radicals. Overall, the results of the present study can be applied for the large-scale production of valuable phenolic acids and flavonoids from P. frutescens through CuNPs and AgNPs 100 mg/L elicited cell suspension cultures.

B-184 Development and Validation of a High-performance Liquid Chromatography-Mass Spectrometry Method for Analyzing Phenolic Acids Metabolites in Feces

Abstract Background Phenolic acids are beneficial metabolites produced by Co-metabolism between a human host and the gastrointestinal microbiota. They play a part in signaling pathways involved in intestinal mucosa homeostasis. The synergic relationship between the microbiota and the nutrient in the gut is associated with the content of phenolic compounds found in the gut system. Recent studies have proven that phenolic acids are linked to weight loss, but such an association needs to be better understood. This research aims to develop and validate a Liquid Chromatography tandem Mass spectrometry (HPLC-MS/MS) method for quantifying phenolic acids in human fecal samples according to the food and Drug Administration (FDA) guidelines. Moreover, the research focuses on the synergic relationship between the gut microbiota and the prebiotics, and their ability to enhance the production of phenolic acids, as an increase in the phenolic acid level in the gut may reduce obesity. Methods High-pressure liquid chromatography is used to separate the analytes (ferulic acid, gallic acid, and caffeic acid) based on polarity, and AB SCIEX QTRAP triple quadrupole Tendon mass spectrometer is used to conduct mass spectroscopy using electrospray ionization. Solid phase extraction method was used to extract and purify the sample with C18 SPE cartridges before analysis. Quantitative analysis was performed by Analyst software. The developed LC-MS/MS method was applied to fecal samples collected from healthy populations. For testing the influence of prebiotics on the production of phenolic acids. Samples of gut microbiota (lactobacillus acidophilus) mixed with different prebiotics were monitored to measure the outcomes of phenolic acids. Samples were compared with other prebiotics such as fiber, cellulose, and inulin. Target analytes meeting significance of (P < 0.05) will be verified by the development of a targeted LC-MS/MS (MRM) assay. Once compounds of significance have been verified, the method will be evaluated based on the guideline from 2018, Bioanalytical method validation guidance for industry; several parameters must be considered when validating a method. These parameters include selectivity, accuracy, precision & recovery, linearity, sensitivity (LOD and LOQ), stability, and matrix effect. Validation will be applied to the stool samples from an average healthy population. Results Phenolic acids were extracted and quantified using a simplified HPLC-MS/MS, with acidified methanol. A total of five phenolic acids with different concentrations were identified in different fecal samples. The method was used to determine the association between gut microbes and nutrient sources and their influence on the production of phenolic acids when mixed. Initial preliminary data indicate that adding prebiotics to a probiotic media increases the peak intensity suggesting an expansion of the quantity of phenolic acid in the mixture. In addition, changing the amount or the type of prebiotics affects the outcomes. Fiber shows the most significant outcomes when compared to other prebiotics. Conclusion Based on this research’s findings, certain types of Prebiotics may enhance the generation of phenolic acid quantity through co-metabolism between a human host and Probiotics in the gut. Fiber showed the most significant outcomes of phenolic acid formation compared to inulin and cellulose.

Long term study of cell wall saccharification and high added value phenolic acids production from Phalaris aquatica L. energy crop

Phalaris aquatica L. is a cool-season perennial grass of the Mediterranean that has been used for forage production, while recently was identified as a novel candidate for sustainable second-generation bioethanol production. This study provides for the first time evidence about this energy crop long-term performance (five years of growth), the cell wall saccharification efficiency and the production of high added value compounds during this period. High biomass production was recorded constantly after the first year of plant growth, with a maximum of 22.4 ± 0.5 t of dry mass (DM) per ha and stable structural polysaccharides concentration 68.8% ± 0.8 w/w DM from the third year on. Independently from crop age, dilute acid pretreatment (120 °C, 1.5% H2SO4, 45 min) promoted the subsequent enzymatic hydrolysis of P. aquatica L. biomass. The dilute acid pre-treatment positive effect is attributed both to the hemicellulose solubilization and the breakage of intermolecular bonds between polysaccharides and phenolic acids, as supported by the present study results These combined acid/enzymatic hydrolysis processes were able to convert more than 80% of cell wall polysaccharides into fermentable sugars, releasing simultaneously a substantial amount of phenolic acids of 2.16% ± 0.48 w/w DM. The ratio of ferulic to p-coumaric acid in the acid hydrolysate can be used for the prediction of the subsequent cellulose enzymatic saccharification efficiency, by the model described here. The protocatechuic acid, which can find many technological applications, was the main bioactive polyphenol in the final hydrolysate with concentration ∼1% w/w DM. These results are supportive of Phalaris aquatica L. suitability as a sustainable biorefinery crop, under the semi-arid Mediterranean climatic conditions contributing to this region biodiversity and prosperity.

Emergence of the Dickeya genus involved duplication of the OmpF porin and the adaptation of the EnvZ-OmpR signaling network.

Genome evolution, and more specifically gene duplication, is a key process shaping host-microorganism interaction. The conserved paralogs usually provide an advantage to the bacterium to thrive. If not, these genes become pseudogenes and disappear. Here, we show that during the emergence of the genus Dickeya, the gene encoding the porin OmpF was duplicated. Our results show that the ompF2 expression is deleterious to the virulence of Dickeya dadantii, the agent causing soft rot disease. Interestingly, ompF2 is regulated while ompF is constitutive but activated by the EnvZ-OmpR two-component system. In vitro, acidic pH triggers the system. The pH measured in four eudicotyledons increased from an initial pH of 5.5 to 7 within 8 h post-infection. Then, the pH decreased to 5.5 at 10 h post-infection and until full maceration of the plant tissue. Yet, the production of phenolic acids by the plant's defenses prevents the activation of the EnvZ-OmpR system to avoid the ompF2 expression even though environmental conditions should trigger this system. We highlight that gene duplication in a pathogen is not automatically an advantage for the infectious process and that, there was a need for our model organism to adapt its genetic regulatory networks to conserve these duplicated genes. IMPORTANCE Dickeya species cause various diseases in a wide range of crops and ornamental plants. Understanding the molecular program that allows the bacterium to colonize the plant is key to developing new pest control methods. Unlike other enterobacterial pathogens, Dickeya dadantii, the causal agent of soft rot disease, does not require the EnvZ-OmpR system for virulence. Here, we showed that during the emergence of the genus Dickeya, the gene encoding the porin OmpF was duplicated and that the expression of ompF2 was deleterious for virulence. We revealed that while the EnvZ-OmpR system was activated in vitro by acidic pH and even though the pH was acidic when the plant is colonized, this system was repressed by phenolic acid (generated by the plant's defenses). These results provide a unique- biologically relevant-perspective on the consequence of gene duplication and the adaptive nature of regulatory networks to retain the duplicated gene.

Efficient production of ferulic acid and p-coumaric acid from reed straws via combined enzymatic hydrolysis and hydrothermal pretreatment

The present study aimed to evaluate an available process for production of ferulic acid (FA) and p-coumaric acid (pCA) that could be potentially used as functional agents in food processing. The hydrothermal pretreatment combined with enzymatic hydrolysis (HP-CEH) was explored for efficient production of phenolic acids from reed straws. The results indicated that HP-CEH significantly increased production of phenolic acids. The recovery yield of ferulic acid and p-coumaric acid was respectively found to be 0.69 and 0.81 mg/g dry reed straw when performed by HP-CEH for 16 h. The degradation of hemicellulose, lignocellulose and lignin, and disruption of cell walls or cellulose fibers contributed to phenolic acids production from reed straws. Moreover, the scavenging activity of hydroxyl radical and DPPH radical of the phenolic acids extracted by HP-CEH was respectively obtained to be 77.32% and 86.77%.

Pomoea batatas(L.) Lam. (Convolvulaceae) as a source of polyphenols with antitumor activity and prospects for in vitro production using chemical elicitors -A Review

Ipomoea batatas has an enormous projection as functional food and it is an excellent source of anticancerous or chemopreventive substances. Plant tissue culture offers the possibility of inducing secondary metabolites production under controlled conditions and their projection for industrial purposes. To establish the state of knowledge regarding advances in polyphenols chemical elicitation from I. batatas and the possibility of producing potential anticancerous compounds in vitro culture systems, a bibliometric review and an analysis of information available until 2020 were made. Results showed that research field on which review was carried out is current and has a growing trend; however, sweet potato cell suspensions evaluation is scarce. Elicitation using compounds with hormonal effects represents a good alternative to increase phenolic acids production in this species. Advances in phenols elicitation from sweet potato cell suspensions will require search superior genotypes, and associating this knowledge with in vivo and in silico studies.

Temporary immersion bioreactors as a useful tool for obtaining high productivity of phenolic compounds with strong antioxidant properties from Pontechium maculatum

Pontechium maculatum (Russian bugloss) is a medical plant belonging to the family Boraginaceae. Although this species is known as a medical plant rich in biologically active secondary metabolites, biotechnological studies about this valuable plant is still missing. The scientific objectives of this study were to investigate the biomass production, synthesis, and productivity of various phenolic acids, flavonoids, and shikonin in P. maculatum cultivated in various breeding systems. Additionally, the antioxidant activity of plant-derived extracts was evaluated. Plants were cultivated in a traditional agar-solidified medium, a liquid medium with rotary shaking, and a temporary immersion bioreactors Plantform™ (TIB), as well as cultivated in soil (ex vitro conditions). Analyses of the growth index and dry weight accumulation were performed on the collected material. In the extracts obtained from examined plants, total phenolic content was estimated, and qualitative and quantitative analysis of phenolic derivatives using DAD-HPLC was conducted, simultaneously with an analysis of antioxidant capacity. TIB stimulated the highest synthesis of all examined phenolic acids and shikonin. In TIB-cultivated shoots level of rosmarinic acid obtained a concentration of 3160.76 mg × 100 g− 1 dry weight (DW), and shikonin obtained a concentration of 77.26 mg × 100 g− 1 DW. Furthermore, plants from TIB were characterized by the highest productivity of all studied phenolic derivatives, what makes it very effective platform for the synthesis of biologically active secondary metabolites in Russian bugloss. Moreover, this article shows that P. maculatum is a rich source of various phenolic derivatives with high antioxidant potential.

The Influence of Exogenous Phenylalanine on the Accumulation of Secondary Metabolites in Agitated Shoot Cultures of Ruta graveolens L.

This study aimed to examine the influence of the addition of a precursor (phenylalanine) on the accumulation of secondary metabolites in agitated shoot cultures of Ruta graveolens. Cultures were grown on Linsmaier and Skoog (LS) medium, with plant growth regulators (0.1 mg/L α-naphthaleneacetic acid-NAA-and 0.1 mg/L 6-benzylaminopurine-BAP). Phenylalanine was added to the cultures at a concentration of 1.25 g/L after 4 and 5 weeks of growth cycles. Biomass was collected after 2, 4, and 7 days of precursor addition. Both control and experimental cultures had the same secondary metabolites accumulated. Using the HPLC method, linear furanocoumarins (bergapten, isoimperatorin, isopimpinellin, psoralen, and xanthotoxin), furoquinoline alkaloids (γ-fagarine, 7-isopentenyloxy-γ-fagarine, and skimmianine), and catechin were detected and quantified in the methanolic extracts. In turn, phenolic acids, such as gallic acid, protocatechuic acid, p-hydroxybenzoic acid, syringic acid, p-coumaric acid, and ferulic acid were detected in hydrolysates. The production of phenolic acids and catechin (1.5-fold) was significantly increased by the addition of precursor, while there was no significant effect on the production of coumarins and alkaloids. The highest total content of phenolic acids (109 mg/100 g DW) was obtained on the second day of phenylalanine addition (the fourth week of growth cycles). The dominant phenolic compounds were p-coumaric acid (maximum content 64.3 mg/100 g DW) and ferulic acid (maximum content 35.6 mg/100 g DW). In the case of catechins, the highest total content (66 mg/100 g DW) was obtained on the third day of precursor addition (the fourth week of growth cycles). This study is the first to document the effect of feeding the culture medium with phenylalanine on the accumulation of bioactive metabolites in in vitro cultures of R. graveolens.

Characterization of procyanidin extracts from hawthorn (Crataegus pinnatifida) in human colorectal adenocarcinoma cell line Caco-2, simulated Digestion, and fermentation identified unique and novel prebiotic properties

The health-promoting activities of procyanidin extracts from hawthorn (HPCs) are closely related to their digestive behaviors, absorption, and colonic metabolism, all of which remain unknown for now and thus hinder further exploration. This study aims to explore the dynamic changes of HPCs during in vitro digestion and fermentation, as well as their Caco-2 permeability, focusing mainly on the interaction between gut microbiota and HPCs. The results showed that the digested HPC samples had characteristic absorption peaks at 280 nm, and there were absorption peaks in the stretching vibration zone, including OH and CC on the benzene ring, which suggested that procyanidins were the main components in HPCs after in vitro digestion. Meanwhile, HPCs had the highest stability in the oral phase. However, the total procyanidin content of HPCs decreased during gastrointestinal digestion, and flavan-3-ol dimers and trimers in HPCs are partially degraded into epicatechin. Uptake of epicatechin (4.07 %), procyanidin B2 (2.15 %), and procyanidin B5 (39.44 %) through Caco-2 monolayer was also observed in HPC treatment, while there was still a large portion of procyanidins that was not absorbed. Subsequent fermentation resulted in a decrease in pH along with the production of short-chain fatty acids (SCFAs), mainly due to the degradation and utilization of HPC, as indicated by a reduction of total procyanidins. Furthermore, the HPCs modulated gut microbial populations: down-regulated the abundances of Bacteroides, Fusobacterium, Enterococcus, Parabacteroides, and Bilophila, and up-regulated Escherichia-Shigella, Klebsiella, Turicibacter, Actinobacillus, Roseburia, and Blautia. Ultimately, epicatechin and procyanidin B2, B5 and C1 were converted into phenolic acids through the metabolism of Bacteroides, Sutterella, Butyrobacter and Blautia. 4-ethylbenzoic acid, 4-hydroxyphenylpropionic acid, 3,4-dihydroxyphenyl acetic acid were confirmed as the significant metabolites in the fermentation. These results elucidated the potential mechanisms of HPCs metabolism and their beneficial effects on gut microbiota and colonic phenolic acids production.

LC-HRMS/MS-Based Metabolomics Approaches Applied to the Detection of Antifungal Compounds and a Metabolic Dynamic Assessment of Orchidaceae.

The liquid chromatography-mass spectrometry (LC-MS)-based metabolomics approach is a powerful technology for discovering novel biologically active molecules. In this study, we investigated the metabolic profiling of Orchidaceae species using LC-HRMS/MS data combined with chemometric methods and dereplication tools to discover antifungal compounds. We analyze twenty ethanolic plant extracts from Vanda and Cattleya (Orchidaceae) genera. Molecular networking and chemometric methods were used to discriminate ions that differentiate healthy and fungal-infected plant samples. Fifty-three metabolites were rapidly annotated through spectral library matching and in silico fragmentation tools. The metabolomic profiling showed a large production of polyphenols, including flavonoids, phenolic acids, chromones, stilbenoids, and tannins, which varied in relative abundance across species. Considering the presence and abundance of metabolites in both groups of samples, we can infer that these constituents are associated with biochemical responses to microbial attacks. In addition, we evaluated the metabolic dynamic through the synthesis of stilbenoids in fungal-infected plants. The tricin derivative flavonoid- and the loliolide terpenoidfound only in healthy plant samples, are promising antifungal metabolites. LC-HRMS/MS, combined with state-of-the-art tools, proved to be a rapid and reliable technique for fingerprinting medicinal plants and discovering new hits and leads.