Aromatic Compounds In Plants Research Articles

Efficient extraction based on a polydimethylsiloxane/bimetallic ZnCo-MOF carbonization sponge coupled with GC-MS for the rapid analysis of volatile compounds in cumin.

A novel porous polydimethylsiloxane/bimetallic ZnCo-MOF carbonization (PDMS/ZnCo-MOF@C) sponge was successfully fabricated, followed by its utilization in GC-MS for the high efficiency extraction and determination of volatile compounds in cumin. The PDMS/ZnCo-MOF@C sponge exhibits outstanding properties with a considerable adsorption capacity, high surface area, and large pore volume and has shown potential as an ideal adsorbent for the separation and preconcentration of trace volatile compounds. The effect of different parameters on the extraction efficiency were investigated. Excellent analytical performances were achieved for the representative compounds (β-pinene, p-cymene, γ-terpinene, cuminaldehyde, and linalyl acetate), including wide linearity (2.31-440.1 ng) with high correlation coefficients (R2 ≥ 0.9979), low LODs (1.02-3.11 ng) and LOQs (2.45-7.08 ng), and satisfactory precision (intra-day RSDs ≤ 2.89% and inter-day RSDs ≤ 4.14%). The optimal method was applied for the analysis of cumin from different regions and 44 volatile compounds were identified. The correlation between the different regions of cumin and volatile compounds was explored using multivariate statistical analysis. These results demonstrated that PDMS/ZnCo-MOF@C is an efficient, simple and sensitive material for use in the pretreatment technique for the determination of the volatile compounds in aromatic plants.

Exploring the Use of Aromatic Compounds in Crop Growth and Protection

In the realm of sustainable agriculture, the integration of aromatic compounds presents a transformative approach to crop growth and protection. This review offers a comprehensive exploration of the role, mechanisms, applications, and future prospects of aromatic compounds in modern agriculture. Derived from both natural and synthetic sources, these compounds have showcased efficacy in enhancing photosynthesis, modulating stress responses in plants, and exhibiting repellent or toxic effects on pests and diseases. Technological innovations, notably in delivery systems like nanoencapsulation and precision agriculture, promise optimized deployment of these compounds. Genetic engineering provides avenues for enhanced in-situ production of aromatic compounds in plants, presenting an intriguing confluence of biology and agronomy. Synergistic research, emphasizing the combined application of multiple aromatic compounds, is revealing amplified benefits, signifying the potential of combinatorial approaches in the field. However, the journey ahead is dotted with challenges. Resistance development against pests, scalability concerns, and regulatory intricacies necessitate vigilant research and proactive strategies. Economic considerations further accentuate the need for cost-effective solutions, balancing initial investments with long-term gains. The environmental and human health implications of aromatic compounds underscore their ecological significance, advocating for their wider adoption in farming practices. With the organic and sustainable farming sectors witnessing exponential growth, the demand and market potential for aromatic compounds are poised for an upward trajectory. Conclusively, this review elucidates the pivotal role of aromatic compounds in ushering an era of eco-friendly, productive, and sustainable agriculture, bridging the traditional knowledge of nature with cutting-edge scientific research. The synthesis of insights presented herein seeks to guide researchers, policymakers, and practitioners in harnessing the myriad benefits of aromatic compounds, charting a greener future for global agriculture.

Polycyclic aromatic compounds in plants and peat in the peatlands of the European part of Russian Arctic

Polycyclic aromatic compounds in plants and peat in the peatlands of the European part of Russian Arctic

Advances in the microbial synthesis of aromatic fragrance molecules

Aromatic compounds make up a large part of fragrances and are traditionally produced by chemical synthesis and direct extraction from plants. Chemical synthesis depends on petroleum resources and has disadvantages such as causing environment pollutions and harsh reaction conditions. Due to the low content of aromatic compounds in plants and the low yield of direct extraction, plant extractions require large amounts of plant resources that occupy arable land. In recent years, with the development of metabolic engineering and synthetic biology, microbial synthesis of aromatic compounds from renewable resources has become a promising alternative approach to traditional methods. This review describes the research progress on the synthesis of aromatic fragrances by model microorganisms such as Escherichia coli or yeast, including the synthesis of vanillin through shikimic acid pathway and the synthesis of raspberry ketone through polyketide pathway. Moreover, this review highlights the elucidation of native biosynthesis pathways, the construction of synthetic pathways and metabolic regulation for the production of aromatic fragrances by microbial fermentation.

Humic substances from green compost increase bioactivity and antibacterial properties of essential oils in Basil leaves

BackgroundThe Essential oils (EOs) are important bioactive secondary metabolites of aromatic plants, such as Basil, and find extensive utilization in pharmaceutical sector as therapeutic agents. A more eco-friendly industrial production of EOs requires new sustainable methods to improve yield and quality of these bioactive compounds in aromatic plants. Here, we evaluated the biostimulation effects of natural organic materials, such as humic substances (HS) extracted from a humified green compost made with artichoke biomasses.ResultsThe molecular characteristics of HS from green compost were determined by 13C-CPMAS NMR spectroscopy and offline pyrolysis–gas chromatography/mass spectrometry and combined to the bioactivity of the EOs extracts to investigate the structural–activity relationship. Basil plants were grown under HS treatments at different concentrations and the yield, the chemical composition and the antioxidant and antimicrobial properties were determined for EOs extracted by steam distillation method from Basil leaves. HS showed a positive effect on total extraction yield of the most bioactive EO components, with a progressive enhancement of abundance for eugenol, eucalyptol and geranyl acetate with increasing HS concentrations. Humic biostimulation influenced the EOs antioxidant proprieties, as well as their antimicrobial activity against some common Gram-negative bacterial strains such as Klebsiella pneumoniae. The HS biostimulation of Basil plants was associated to their structural hydrophobic characteristics that may have favoured the interactions with the Basil rhizome and to specific bioactive molecular components, such as a lignin-derived phenols and polar saccharidic and aminoacidic compounds.ConclusionsOur results support the potential use of humic substances from green compost as promising effective biostimulants to improve the production and bioactive properties of EOs in aromatic plants.Graphic

Effect of temperature and cut size on the volatile organic compound profile, and expression ofChorismate synthasein fresh-cut melon

The postharvest quality of fresh-cut melon is strongly affected by storage conditions to which it is subjected. During postharvest, fruit undergoes several stresses and its physiology is similar to that in senescent tissues. This affects both its biochemistry and the expression of genes involved in secondary metabolite biosynthesis. Volatile organic compounds (VOCs) have been used previously to assess quality of fresh cut melon as they reflect changes in flavor and also changes in overall metabolism. Chorismate synthase (CS) is a key enzyme in the shikimate pathway and catalyzes the formation of chorismate, which is the precursor of numerous aromatic compounds in plants. In this work the effects of different storage temperatures and cut-sizes were studied, with the aim of identifying effects on flavor through changes in VOCs, and molecular responses of the CmCS gene to different postharvest conditions. Melon (Cucumis melo L. 'Macigno') fruits were harvested at a fully ripened commercial stage, were washed in a chlorine water solution, and the mesocarp (pulp) was cut in cube-shaped portions; two sizes were chosen, 1×1 and 3×2 cm. Melon cubes were then stored at 20 or 4°C. VOCs were assessed in the 3×2 cut size at both temperatures and showed clear changes during storage. A subset of VOCs were shown to correlate negatively with storage time and temperature and from these, specific compounds can be identified that act as markers for an overall change in VOC profiles. CmCS was more affected by temperature, showing decreased levels of expression during storage at 20°C with respect to harvest and to cold storage. On the other hand, cut-size did not result in changes in its expression in 3×2 cut size.

UHPLC-DAD Multi-Method for Determination of Phenolics in Aromatic Plants

The aim of this paper was to develop and validate a method of ultra high-performance liquid chromatography coupled with diode array detector (UHPLC-DAD), which allows quantifying simultaneously 19 phenolic compounds in aromatic plants. Twenty-one fresh and dried aromatic plants were extracted with methanol before their UHPLC-DAD analysis. The method was validated regarding specificity, working range, linearity, limit of detection (LOD) and quantification (LOQ), precision, and accuracy. The developed method allows detecting and quantifying the phenolic compounds at low concentrations (LOD < 0.15 μg/mL, except for carnosol and carnosic acid). The relative standard deviation obtained was lower than 5.73 and 8.75%, for repeatability and intermediate precision, respectively. The proposed method is straightforward and sensitive, with good precision and accuracy. Dried thyme presented the highest diversity of phenolic compounds and rosmarinic acid was present in most of the samples. This analytical tool is a valid alternative to conventional methods to quantify phenolic compounds in aromatic plants or in their extracts.

Characterization of a class II 5-enopyruvylshikimate-3-phosphate synthase with high tolerance to glyphosate from Sinorhizobium fredii.

5-Enopyruvylshikimate-3-phosphate synthase (EPSP synthase) is an important enzyme in the shikimate pathway mediating the biosynthesis of aromatic compounds in plants and microorganisms. A novel class II EPSP synthase AroA S. fredii from Sinorhizobium fredii NGR234 was overexpressed in Escherichia coli BL21. It was purified to homogeneity and its catalytic properties were studied. The enzyme exhibited optimum catalytic activity at pH 8.0 and 50 °C. It was stable below 40 °C, and over a broad range of pH 5.0-9.0. The EPSP synthase was increasingly activated by 100 mM of the chlorides of NH4 (+), K(+), Na(+) and Li(+). Kinetic analysis of AroA S. fredii suggested that the enzyme exhibited a high glyphosate tolerance and high level of affinity for phosphoenolpyruvate, which indicates the enzyme with a high potential for structural and functional studies and its potential usage for the generation of transgenic crops resistant to the herbicide.

Eichhornia crassipes: An advantageous source of shikimic acid

The water hyacinth (Eichhornia crassipes (Mart.) Solms, Pontederiaceae) is considered as one of the most productive plants on earth, and an aquatic weed, which causes serious environmental problems. In this study, this species is presented as an alternative of a renewable source of shikimic acid. Although this acid is an important intermediate in the biosynthesis of aromatic compounds in plants and microorganisms, its occurrence is described for the first time in a species of the Pontederiaceae family. Shikimic acid is the lead compound for the production of the antiviral agent oseltamivir phosphate (Tamiflu®). Semi-quantitative analyses of the plant extracts by HPLC-PDA showed that the aerial parts of E. crassipes contain higher shikimic acid concentration (0.03%-2.70% w/w) than the roots (0.05%-0.90% w/w), and that methanol is a better solvent than water for shikimic acid extraction.

Analysis of Naturally Occurring Phenolic Compounds in Aromatic Plants by RP-HPLC Coupled to Diode Array Detector (DAD) and GC-MS after Silylation.

The following aromatic plants of Greek origin, Origanum dictamnus (dictamus), Eucalyptus globulus (eucalyptus), Origanum vulgare L. (oregano), Mellisa officinalis L. (balm mint) and Sideritis cretica (mountain tea), were examined for the content of phenolic substances. Reversed phase HPLC coupled to diode array detector (DAD) was used for the analysis of the plant extracts. The gas chromatography-mass spectrometry method (GC-MS) was also used for identification of phenolic compounds after silylation. The most abundant phenolic acids were: gallic acid (1.5–2.6 mg/100 g dry sample), ferulic acid (0.34–6.9 mg/100 g dry sample) and caffeic acid (1.0–13.8 mg/100 g dry sample). (+)-Catechin and (−)-epicatechin were the main flavonoids identified in oregano and mountain tea. Quercetin was detected only in eucalyptus and mountain tea.

Heterologous expression and characterization of soluble recombinant 3-deoxy-d-arabino-heptulosonate-7-phosphate synthase from Actinosynnema pretiosum ssp. auranticum ATCC31565 through co-expression with Chaperones in Escherichia coli

3-Deoxy-d-arabino-heptulosonate-7-phosphate synthase (DAHPS), (EC 2.5.1.54) catalyzes the first step of the shikimate pathway, the route for the biosynthesis of aromatic compounds in plants and microbes. In Actinosynnema pretiosum, the aroF gene (GenBank: AF056968.1) encodes DAHPS to condensate phosphoenolpyruvate (PEP) and d-erythrose 4-phosphate (E4P) to generate DAHP. In this study, a recombinant pET28a-aroF plasmid was constructed and A. pretiosum DAHPS was successfully expressed in soluble form by co-expression with chaperonins GroEL/GroES in Escherichia coli. The purification and kinetic characterization of the expressed protein were then investigated. The DAHPS originated from A. pretiosum demonstrated a pronounced substrate inhibition by PEP but was not sensitive to E4P. The purified enzyme was completely inactivated by EDTA but potently activated by several bivalent metal ions, especially Mn2+ and Co2+.

β-Glucosidases.

β-Glucosidases (3.2.1.21) are found in all domains of living organisms, where they play essential roles in the removal of nonreducing terminal glucosyl residues from saccharides and glycosides. β-Glucosidases function in glycolipid and exogenous glycoside metabolism in animals, defense, cell wall lignification, cell wall β-glucan turnover, phytohormone activation, and release of aromatic compounds in plants, and biomass conversion in microorganisms. These functions lead to many agricultural and industrial applications. β-Glucosidases have been classified into glycoside hydrolase (GH) families GH1, GH3, GH5, GH9, and GH30, based on their amino acid sequences, while other β-glucosidases remain to be classified. The GH1, GH5, and GH30 β-glucosidases fall in GH Clan A, which consists of proteins with (β/α)(8)-barrel structures. In contrast, the active site of GH3 enzymes comprises two domains, while GH9 enzymes have (α/α)(6) barrel structures. The mechanism by which GH1 enzymes recognize and hydrolyze substrates with different specificities remains an area of intense study.

Structural Basis of Glyphosate Tolerance Resulting from Mutations of Pro101 in Escherichia coli 5-Enolpyruvylshikimate-3-phosphate Synthase

Glyphosate, the world's most used herbicide, is a massive success because it enables efficient weed control with minimal animal and environmental toxicity. The molecular target of glyphosate is 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), which catalyzes the sixth step of the shikimate pathway in plants and microorganisms. Glyphosate-tolerant variants of EPSPS constitute the basis of genetically engineered herbicide-tolerant crops. A single-site mutation of Pro(101) in EPSPS (numbering according to the enzyme from Escherichia coli) has been implicated in glyphosate-resistant weeds, but this residue is not directly involved in glyphosate binding, and the basis for this phenomenon has remained unclear in the absence of further kinetic and structural characterization. To probe the effects of mutations at this site, E. coli EPSPS enzymes were produced with glycine, alanine, serine, or leucine substituted for Pro(101). These mutant enzymes were analyzed by steady-state kinetics, and the crystal structures of the substrate binary and substrate.glyphosate ternary complexes of P101S and P101L EPSPS were determined to between 1.5- and 1.6-A resolution. It appears that residues smaller than leucine may be substituted for Pro(101) without decreasing catalytic efficiency. Any mutation at this site results in a structural change in the glyphosate-binding site, shifting Thr(97) and Gly(96) toward the inhibitor molecule. We conclude that the decreased inhibitory potency observed for glyphosate is a result of these mutation-induced long-range structural changes. The implications of our findings concerning the development and spread of glyphosate-resistant weeds are discussed.

Characterization of a recombinant type II 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase from Helicobacter pylori

DAH7P (3-Deoxy-D-arabino-heptulosonate 7-phosphate) synthase catalyses the condensation reaction between phosphoenolpyruvate (PEP) and D-erythrose 4-phosphate (E4P) as the first committed step in the biosynthesis of aromatic compounds in plants and micro-organisms. Previous work has identified two families of DAH7P synthases based on sequence similarity and molecular mass, with the majority of the mechanistic and structural studies being carried out on the type I paralogues from Escherichia coli. Whereas a number of organisms possess genes encoding both type I and type II DAH7P synthases, the pathogen Helicobacter pylori has only a single, type II, enzyme. Recombinant DAH7P synthase from H. pylori was partially solubilized by co-expression with chaperonins GroEL/GroES in E. coli, and purified to homogeneity. The enzyme reaction follows an ordered sequential mechanism with the following kinetic parameters: K(m) (PEP), 3 microM; K(m) (E4P), 6 microM; and kcat, 3.3 s(-1). The enzyme reaction involves interaction of the si face of PEP with the re face of E4P. H. pylori DAH7P synthase is not inhibited by phenylalanine, tyrosine, tryptophan or chorismate. EDTA inactivates the enzyme, and activity is restored by a range of bivalent metal ions, including (in order of decreasing effectiveness) Co2+, Mn2+, Ca2+, Mg2+, Cu2+ and Zn2+. Analysis of type II DAH7P synthase sequences reveals several highly conserved motifs, and comparison with the type I enzymes suggests that catalysis by these two enzyme types occurs on a similar active-site scaffold and that the two DAH7P synthase families may indeed be distantly related.

Determination of phenolic compounds in aromatic plants by RP-HPLC and GC-MS

It is well known that phenolic compounds are constituents of many plants and herbs, and they have attracted a great deal of public and scientific interest because of their health-promoting effects as antioxidants. Five plants, Vitex agnus-castus (Verbenaceae), Origanum dictamnus (Lamiaceae), Teucrium polium (Lamiaceae), Lavandula vera (Lamiaceae) and Lippia triphylla (Verbenaceae), were examined in order to determine their phenolic composition. Reversed phase high performance liquid chromatography was employed for the identification and quantification of phenolic compounds. Gas chromatography–mass spectometry(GC-MS) was also used for identification of phenolic compounds after silylation. Analysis of the non-volatile and thermolabile phenolic compounds by GC-MS presupposes their conversion into volatile and thermotolerant derivatives. The derivatization process was optimized against reagents, temperature and reaction time. A large excess of N, O-bis(trimethylsilyl)trifluoroacetamide containing trimethylchlorosilane proved to be the best derivatization reagent to convert analytes into volatile trimethylsilyl derivatives. The most abundant phenolic compounds detected were caffeic acid (0.12–0.93 mg 100 g −1 dry sample), ferulic acid (0.34–1.52 mg 100 g −1 dry sample), and (+)-catechin (0.22–0.43 mg 100 g −1 dry sample).

Degradation of aromatic compounds in plants grown under aseptic conditions.

The aim of the work is to investigate the ability of higher plants to absorb and detoxify environmental pollutants - aromatic compounds via aromatic ring cleavage. Transformation of 14C specifically labelled benzene derivatives, [1-6-14C]-nitrobenzene, [1-6-(14)C]-aniline, [1-(14)C]- and [7-(14)C]-benzoic acid, in axenic seedlings of maize (Zea mays L.), kidney bean (Phaseolus vulgaris L.), pea (Pisum sativum L.) and pumpkin (Cucurbita pepo L.) were studied. After penetration in plants, the above xenobiotics are transformed by oxidative or reductive reactions, conjugation with cell endogenous compounds, and binding to biopolymers. The initial stage of oxidative degradation consists in hydroxylation reactions. The aromatic ring can then be cleaved and degraded into organic acids of the Krebs cycle. Ring cleavage is accompanied by 14CO2 evolution. Aromatic ring cleavage in plants has thus been demonstrated for different xenobiotics carrying different substitutions on their benzene ring. Conjugation with low molecular peptides is the main pathway of aromatic xenobiotics detoxification. Peptide conjugates are formed both by the initial xenobiotics (except nitrobenzene) and by intermediate transformation products. The chemical nature of the radioactive fragment and the amino acid composition of peptides participating in conjugation were identified.

Molecular models for shikimate pathway enzymes of Xylella fastidiosa

The Xylella fastidiosa is a bacterium that is the cause of citrus variegated chlorosis (CVC). The shikimate pathway is of pivotal importance for production of a plethora of aromatic compounds in plants, bacteria, and fungi. Putative structural differences in the enzymes from the shikimate pathway, between the proteins of bacterial origin and those of plants, could be used for the development of a drug for the control of CVC. However, inhibitors for shikimate pathway enzymes should have high specificity for X. fastidiosa enzymes, since they are also present in plants. In order to pave the way for structural and functional efforts towards antimicrobial agent development, here we describe the molecular modeling of seven enzymes of the shikimate pathway of X. fastidiosa. The structural models of shikimate pathway enzymes, complexed with inhibitors, strongly indicate that the previously identified inhibitors may also inhibit the X. fastidiosa enzymes.

Conservation of mechanism in three chorismate-utilizing enzymes.

Chorismate is the end-product of the shikimate pathway for biosynthesis of carbocyclic aromatic compounds in plants, bacteria, fungi, and some parasites. Anthranilate synthase (AS), 4-amino-4-deoxychorismate synthase (ADCS), and isochorismate synthase (IS) are homologous enzymes that carry out the initial transformations on chorismate in the biosynthesis of tryptophan, p-aminobenzoate, and enterobactin, respectively, and are expected to share a common mechanism. Poor binding to ADCS of two potential transition state analogues for addition of a nucleophile to C6 of chorismate implies that it, like AS and IS, initiates reaction by addition of a nucleophile to C2. Molecular modeling based on the X-ray structures of AS and ADCS suggests that the active site residue K274 is the nucleophile employed by ADCS to initiate the reaction, forming a covalent intermediate. The K274A and K274R mutants were shown to have 265- and 640-fold reduced k(cat) values when PabA (the cognate amidotransferase) + glutamine are used as the nitrogen source. Under conditions of saturating chorismate and NH(4)(+), ADCS and the K274A mutant have identical k(cat) values, suggesting the participation of NH(4)(+) as a rescue agent. Such participation was confirmed by the buildup of 2-amino-2-deoxyisochorismate in the reactions of the K274A mutant but not ADCS, when either NH(4)(+) or PabA + glutamine is used as the nitrogen source. Additionally, the inclusion of ethylamine in the reactions of K274A yields the N-ethyl derivative of 2-amino-2-deoxyisochorismate. A unifying mechanism for AS, ADCS, and IS entailing nucleophile addition to C2 of chorismate in an S(N)2' ' process is proposed.

Structures of Shikimate Dehydrogenase AroE and Its Paralog YdiB: A COMMON STRUCTURAL FRAMEWORK FOR DIFFERENT ACTIVITIES

Shikimate dehydrogenase catalyzes the fourth step of the shikimate pathway, the essential route for the biosynthesis of aromatic compounds in plants and microorganisms. Absent in metazoans, this pathway is an attractive target for nontoxic herbicides and drugs. Escherichia coli expresses two shikimate dehydrogenase paralogs, the NADP-specific AroE and a putative enzyme YdiB. Here we characterize YdiB as a dual specificity quinate/shikimate dehydrogenase that utilizes either NAD or NADP as a cofactor. Structures of AroE and YdiB with bound cofactors were determined at 1.5 and 2.5 A resolution, respectively. Both enzymes display a similar architecture with two alpha/beta domains separated by a wide cleft. Comparison of their dinucleotide-binding domains reveals the molecular basis for cofactor specificity. Independent molecules display conformational flexibility suggesting that a switch between open and closed conformations occurs upon substrate binding. Sequence analysis and structural comparison led us to propose the catalytic machinery and a model for 3-dehydroshikimate recognition. Furthermore, we discuss the evolutionary and metabolic implications of the presence of two shikimate dehydrogenases in E. coli and other organisms.

How the mutation glycine96 to alanine confers glyphosate insensitivity to 5-enolpyruvyl shikimate-3-phosphate synthase from Escherichia coli.

The enzyme 5-enolpyruvyl shikimate-3-phosphate (EPSP) synthase (EC 2.5.1.19) is essential for the biosynthesis of aromatic compounds in plants and microbes and is the unique target of the herbicide glyphosate. One of the first glyphosate-insensitive enzymes reported was a Gly96Ala mutant of EPSP synthase from Klebsiella pneumoniae. We have introduced this single-site mutation into the highly homologous EPSP synthase from Escherichia coli. The mutant enzyme is insensitive to glyphosate with unaltered affinity for its first substrate, shikimate-3-phosphate (S3P), but displays a 30-fold lower affinity for its second substrate, phosphoenolpyruvate (PEP). Using X-ray crystallography, we solved the structure of Gly96Ala-EPSP synthase liganded with S3P to 0.17 nm resolution. The crystal structure shows that the additional methyl group from Ala96 protrudes into the active site of the enzyme. While the interactions between enzyme and S3P remain unaffected, the accessible volume for glyphosate binding is substantially reduced. Exploiting the crystallographic results for molecular modeling, we demonstrate that PEP but not glyphosate can be docked in the Gly96Ala-modified binding site. The predicted PEP binding site satisfies the earlier proposed interaction pattern for PEP with EPSP synthase and corroborates the assumption that glyphosate and PEP target the same binding site.