coniferyl-alcohol and ferulic-acid

During the past few years, the production of natural value-added compounds from microbial sources has gained tremendous importance. Due to an increase in consumer demand for natural products, various food and pharmaceutical industries are continuously in search of novel metabolites obtained from microbial biotransformation. The exploitation of microbial biosynthetic pathways is both feasible and cost effective in the production of natural compounds. The environmentally compatible nature of these products is one major reason for their increasing demand. Novel approaches for natural product biogeneration will take advantage of the current studies on biotechnology, biochemical pathways and microbiology. The interest of the scientific community has shifted toward the use of microbial bioconversion for the production of valuable compounds from natural substrates. The present review focuses on eugenol biotransformation by microorganisms resulting in the formation of various value-added products such as ferulic acid, coniferyl alcohol, vanillin and vanillic acid.

Topics: Benzaldehydes; Biological Products; Biotechnology; Biotransformation; Coumaric Acids; Eugenol; Phenols; Vanillic Acid

(Hydroxy)cinnamyl alcohols and allylphenols, including coniferyl alcohol and eugenol, are naturally occurring aromatic compounds widely utilised in pharmaceuticals, flavours, and fragrances. Traditionally, the heterologous biosynthesis of (hydroxy)cinnamyl alcohols from (hydroxy)cinnamic acids involved CoA-dependent activation of the substrate. However, a recently explored alternative pathway involving carboxylic acid reductase (CAR) has proven efficient in generating the (hydroxy)cinnamyl aldehyde intermediate without the need for CoA activation. In this study, we investigated the application of the CAR pathway for whole-cell bioconversion of a range of (hydroxy)cinnamic acids into their corresponding (hydroxy)cinnamyl alcohols. Furthermore, we sought to extend the pathway to enable the production of a variety of allylphenols and allylbenzene.. By screening the activity of several heterologously expressed enzymes in crude cell lysates, we identified the combination of Segniliparus rugosus CAR (SrCAR) and Medicago sativa cinnamyl alcohol dehydrogenase (MsCAD2) as the most efficient enzymatic cascade for the two-step reduction of ferulic acid to coniferyl alcohol. To optimise the whole-cell bioconversion in Escherichia coli, we implemented a combinatorial approach to balance the gene expression levels of SrCAR and MsCAD2. This optimisation resulted in a coniferyl alcohol yield of almost 100%. Furthermore, we extended the pathway by incorporating coniferyl alcohol acyltransferase and eugenol synthase, which allowed for the production of eugenol with a titre of up to 1.61 mM (264 mg/L) from 3 mM ferulic acid. This improvement in titre surpasses previous achievements in the field employing a CoA-dependent coniferyl alcohol biosynthesis pathway. Our study not only demonstrated the successful utilisation of the CAR pathway for the biosynthesis of diverse (hydroxy)cinnamyl alcohols, such as p-coumaryl alcohol, caffeyl alcohol, cinnamyl alcohol, and sinapyl alcohol, from their corresponding (hydroxy)cinnamic acid precursors but also extended the pathway to produce allylphenols, including chavicol, hydroxychavicol, and methoxyeugenol. Notably, the microbial production of methoxyeugenol from sinapic acid represents a novel achievement.. The combination of SrCAR and MsCAD2 enzymes offers an efficient enzymatic cascade for the production of a wide array of (hydroxy)cinnamyl alcohols and, ultimately, allylphenols from their respective (hydroxy)cinnamic acids. This expands the range of value-added molecules that can be generated using microbial cell factories and creates new possibilities for applications in industries such as pharmaceuticals, flavours, and fragrances. These findings underscore the versatility of the CAR pathway, emphasising its potential in various biotechnological applications.

Topics: Eugenol; Pharmaceutical Preparations

Topics: Acids; Arabinose; Cell Wall; Coumaric Acids; Dimerization; Esters; Hydrolysis; Lignin; Phenols; Poaceae; Zea mays

Aging deteriorates vascular functions such as vascular reactivity and stiffness. Thus far, various reports suggest that bioactive compounds can improve vascular functions. However, few age-related studies of natural bioactive compounds are available. The present study attempted to evaluate age-related vasorelaxation of bioactive cinnamic acids, caffeic acid, and ferulic acid using aged rat thoracic aorta. Vasorelaxation was evaluated in thoracic aorta from both 8, 18, and 40 weeks old Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) respectively. The result indicated that caffeic acid possessed the vasorelaxation regardless of aging in WKY and SHR. Moreover, the vasorelaxation of ferulic acid enhanced with aging in SHR. The vasorelaxation behavior was acted in an endothelium-independent manner. To access structure importance of enhanced vasorelaxation, analogues of ferulic acid were tested. In 40 weeks old SHR, 3,4-dimethoxycinnamic acid and coniferyl alcohol exhibited equivalent vasorelaxation activity with ferulic acid, providing the structural importance of methoxy-modified 3-position on the phenyl ring and 2-propenoic moiety. These results firstly demonstrated that enhanced vasorelaxation of ferulic acid with aging and 3,4-dimethoxycinnamic acid and coniferyl alcohol, along with ferulic acid, might exhibit the therapeutic potential of vasoactive power with aging.

Topics: Aging; Animals; Aorta, Thoracic; Caffeic Acids; Cinnamates; Coumaric Acids; Endothelium, Vascular; Male; Phenols; Rats; Rats, Inbred SHR; Rats, Inbred WKY; Vasodilation

Stable isotope-labeled precursors were synthesized for an analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to elucidate the biosynthetic flow of capsaicinoids, capsinoids, and capsiconinoids. [1'-(13)C][5-(2)H]-Vanillin was prepared by the condensation of guaiacol with [(13)C]-chloroform and a D(2)O treatment. Labeled vanillylamine, vanillyl alcohol, ferulic acid, and coniferyl alcohol were prepared from the labeled vanillin. The labeled vanillylamine was converted to labeled capsaicinoid in a crude enzyme solution extracted from pungent Capsicum fruits.

Topics: Benzaldehydes; Benzyl Alcohols; Benzylamines; Capsaicin; Capsicum; Carbon Isotopes; Chemistry, Organic; Chloroform; Chromatography, Liquid; Coumaric Acids; Deuterium Oxide; Guaiacol; Isotope Labeling; Phenols; Plant Extracts; Prodrugs; Tandem Mass Spectrometry

The phenylpropanoid pathway plays important roles in plants following exposure to environmental stresses, such as wounding and pathogen attack, which lead to the production of a variety of compounds, including lignin, flavonoids and phytoalexins. Ferulate 5-hydroxylase (F5H) is a cytochrome P450-dependent monooxygenase that catalyses the hydroxylation of ferulic acid, coniferaldehyde and coniferyl alcohol, leading to sinapic acid and syringyl lignin biosynthesis. We isolated F5H cDNA and genomic DNA from Camptotheca acuminata and investigated the expression pattern of the C. acuminata F5H (CaF5H1) gene in response to wounding. A search against the BLOCKS database of conserved protein motifs indicated that CaF5H1 retains features in common with F5Hs reported from other plants. 5'-flanking region analysis using the PLACE database showed that putative regulatory elements related to various abiotic and biotic stresses, such as drought, wounding, low temperature and pathogens, exist in the 5'-flanking region of CaF5H1. Based upon these analysis results, we investigated the expression pattern of CaF5H1 gene in response to wounding and stress-related molecules. Here, we show that CaF5H1 transcripts accumulated in the leaves in response to mechanical wounding or the application of molecules involved in the stress response, such as ethylene, ABA and hydrogen peroxide (H2O2). The application of salicylic acid and diphenylene iodonium (DPI) inhibited the wound-induced expression of CaF5H1. Taken together, we suggest that wound-induced expression of CaF5H1 may be mediated by MJ and H2O2 and enhanced phenylpropanoid contents via CaF5H1 maybe function in response to various stresses, including wounding, in plants.

Topics: Abscisic Acid; Acrolein; Amino Acid Sequence; Base Sequence; Camptotheca; Coumaric Acids; Cyclopentanes; Cytochrome P-450 Enzyme System; Ethylenes; Hydrogen Peroxide; Mixed Function Oxygenases; Molecular Sequence Data; Onium Compounds; Oxylipins; Phenols; Plant Leaves; Plant Roots; Plant Stems; Salicylic Acid; Sequence Alignment; Transcription, Genetic

Strain NL15-2K was isolated from soil by screening for bacteria capable of catabolizing lignin-related aromatic acids. This isolate was identified as a Streptomyces sp. on the basis of morphology and an analysis of its 16S rRNA gene sequence. NL15-2K utilized caffeic acid, coniferyl alcohol, ferulic acid, isovanillic acid, protocatechuic acid, vanillic acid, vanillin, and veratric acid as sole carbon sources.

Topics: Benzaldehydes; Biodegradation, Environmental; Biotechnology; Caffeic Acids; Carbon; Coumaric Acids; Hydroxybenzoates; Lignin; Microscopy, Electron, Scanning; Models, Chemical; Phenols; Streptomyces; Vanillic Acid

Plant cell walls containing suberin or lignin in the human diet are conjectured to protect against colon cancer. To confirm the existence of authentic lignin in cereal grain dietary fibers, the DFRC (derivatization followed by reductive cleavage) method was applied to different cereal grain dietary fibers. By cleavage of diagnostic arylglycerol-beta-aryl (beta-O-4) ether linkages and identification of the liberated monolignols, it was ascertained that lignins are truly present in cereal grains. From the ratios of the liberated monolignols coniferyl alcohol and sinapyl alcohol, it is suggested that lignin compositions vary among cereals. Furthermore, dimeric cross-coupling products, comprising ferulate and coniferyl alcohol, were identified in most cereal fibers investigated. These ferulate 4-O-beta- and 8-beta-coniferyl alcohol cross-coupled structures indicate radical cross-coupling of polysaccharides to lignin precursors via ferulate.

Topics: Coumaric Acids; Dietary Fiber; Edible Grain; Gas Chromatography-Mass Spectrometry; Lignin; Phenols

Ferulate and diferulates mediate cell wall cross-linking in grasses, but little is known about their cross-coupling reactions with monolignols and their role in lignin formation in primary cell walls. Feruloylated primary walls of maize were artificially lignified and then saponified to release ferulate and diferulates and their cross-products with coniferyl alcohol for analysis by GC-FID, GC-MS, and NMR spectroscopy. Ferulate and 5-5-coupled diferulate had a greater propensity than 8-coupled diferulates to copolymerize with coniferyl alcohol, forming mostly 4-O-beta' and 8-beta' and some 8-O-4' and 8-5' cross-coupled structures. Some 8-beta' structures de-esterified from xylans, but these cross-links were subsequently replaced as 8-coupled diferulates formed stable cross-coupled structures with lignin. Based on the incorporation kinetics of ferulate and diferulates and the predicted growth of lignin, cross-products formed at the onset of lignification acted as nucleation sites for lignin polymerization.

Topics: Cell Wall; Chromatography, Gas; Coumaric Acids; Gas Chromatography-Mass Spectrometry; Lignin; Magnetic Resonance Spectroscopy; Phenols; Saponins; Zea mays

Reactions of barley peroxidase 1 were studied using transient-state and steady-state kinetics at pH 3.96, 25 degrees C, and 0.1 M ionic strength, in both the presence and the absence of 1 mM calcium ion. The rate of compound I formation from barley peroxidase 1 and hydrogen peroxide in the absence of reducing substrate is very slow, with or without calcium. When each of the three reducing substrates ferulic acid, caffeic acid, and coniferyl alcohol is added individually, there is a striking enhancement of the rate of compound I formation by a factor of 10-40 depending on the substrate. These unique rate enhancements can be explained by the effect of tight substrate binding to the native enzyme, and they may be indicative of an activating effect of reducing substrate on barley peroxidase 1 under physiological conditions. All steady-state kinetic results can be explained by an initial tight binding of reducing substrate AH to the barley peroxidase, Peroxidase + AH reversible Peroxidase-AH, and substitution of the peroxidase-AH complex for native enzyme in the standard modified ping-pong mechanism for peroxidase reactions [Dunford, H. B. (1991) in Peroxidases in Chemistry and Biology (Everse, J., Everse, K. E., & Grisham, M. B., Eds.) Vol. II, pp 1-24, CRC Press, Boca Raton, FL]. The dissociation constant of barley peroxidase 1 and ferulic acid was 1.4 +/- 0.6 microM as determined by the change in the absorbance at the Soret maximum at the conditions mentioned above.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Caffeic Acids; Calcium; Coumaric Acids; Hordeum; Hydrogen Peroxide; Kinetics; Mathematics; Models, Theoretical; Peroxidases; Phenols; Plant Proteins; Spectrophotometry; Time Factors

Coniferyl alcohol was shown to be completely biodegradable to carbon dioxide and methane under strictly anaerobic culture conditions. The mineralization of 300 mg of the substrate per liter was observed in acclimated ferulic acid-degrading methanogenic consortia, as well as in anaerobic enrichments on coniferyl alcohol seeded with sewage sludge. Ferulic and phenylpropionic acids were detected in the cultures degrading coniferyl alcohol as the sole carbon and energy source, suggesting that this compound is oxidized to ferulic acid, which is then degraded as previously described.

Topics: Anaerobiosis; Biodegradation, Environmental; Cinnamates; Coumaric Acids; Euryarchaeota; Phenols; Sewage