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Production of phenol oxidases by Paecilomyces variotii and Pestalotia palmarum on solid malt extract medium
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Ferulic acid metabolism was studied in cultures of two micromycetes producing different amounts of phenol oxidases. In cultures of the low phenol oxidase producer Paecilomyces variotii, ferulic acid was decarboxylated to 4-vinylguaiacol, which was converted to vanillin and then either oxidized to vanillic acid or reduced to vanillyl alcohol. Vanill...
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Citations
... The highest occupied molecular orbital (HOMO) and the lowest unoccupied molecular orbital (LUMO) calculations were carried out to determine the reactivity character [38][39][40]. [41] as the carboxylic acid carbon was not that of acid but an ester observed at ẟ C 167.7 ppm; hence, it must be esterified. The second part of the spectrum was typical of a diterpene of the 8, (17) (H-16), the trans-coupled olefinic protons, and the aromatic protons of the ABX spin system. ...
... The compound is not an artefact as the labdane diterpene moiety has not been previously isolated from the plant, and methanol was not used at any point of the isolation and characterization of assay process and procedures; hence, the possibility of methylation by methanol does not exist. Other compounds isolated were the phenolic acids; p-coumaric acid 2 (phenolic extractives from the root bark of Picea abies) [41], ferulic acid 3 (metabolism of ferulic acid by Paecilomyces variotii and Pestalotia palmarum) [41], the fatty acid stearic acid 4 (determination of the fatty acid profile by 1 H NMR spectroscopy) [43], and the triterpenes, lupeol 5 [44] (assignment of 1 H and 13 C spectra and investigation of hindered sidechain rotation in lupeol derivatives), and a mixture of sitosterol and stigmasterol 6 and 7 (isolation of stigmasterol and sitosterol from the dichloromethane extract of Rubus suavissimus) [45]. Their identities were determined and confirmed by comparison to the literature reports and data cited against the compounds. ...
... The compound is not an artefact as the labdane diterpene moiety has not been previously isolated from the plant, and methanol was not used at any point of the isolation and characterization of assay process and procedures; hence, the possibility of methylation by methanol does not exist. Other compounds isolated were the phenolic acids; p-coumaric acid 2 (phenolic extractives from the root bark of Picea abies) [41], ferulic acid 3 (metabolism of ferulic acid by Paecilomyces variotii and Pestalotia palmarum) [41], the fatty acid stearic acid 4 (determination of the fatty acid profile by 1 H NMR spectroscopy) [43], and the triterpenes, lupeol 5 [44] (assignment of 1 H and 13 C spectra and investigation of hindered sidechain rotation in lupeol derivatives), and a mixture of sitosterol and stigmasterol 6 and 7 (isolation of stigmasterol and sitosterol from the dichloromethane extract of Rubus suavissimus) [45]. Their identities were determined and confirmed by comparison to the literature reports and data cited against the compounds. ...
A new labdane diterpene characterized as 18α-O-trans-p-feruloyl-15-methyl-8(17)-labdanoate has been isolated from the roots of Vachellia nilotica. Also isolated were p-coumaric acid, ferulic acid, stearic acid, lupeol, and a mixture of β-sitosterol and stigmasterol. The compounds were obtained after a series of column chromatography on silica gel, and their structures were elucidated using NMR and LC-MS analyses. The new diterpene showed good anti-parasitic activity with an IC50 of 0.0177 µM against Trypanosoma brucei and 0.0154 µM against Leishmania major using an Alamar Blue assay. The compound also displayed very good inhibitory activity against Leishmania major compared to Trypanosoma brucei rhodesiense with a binding energy of −10.5 and −7.8 kcal/mol, respectively. Density functional theory analysis showed that the studied compound has low LUMO–HOMO energy, signifying a high chemical reactivity with the ability to donate electrons to electron-accepting species.
... To the best of our knowledge, this is the first-time vanillin has been detected in P. camemberti fermentation, suggesting this fungus is capable of producing this aroma compound from a precursor available in olive cake (Table 1, Figure 2). Many filamentous fungi are capable of biosynthesising vanillin from different phenolic acids, including P. cinnabarinus [31][32][33]. However, our strain of P. cinnabarinus did not produce this aroma compound in any fermentation. ...
Increasing consumer demand for natural flavours and fragrances has driven up prices and increased pressure on natural resources. A shift in consumer preference towards more sustainable and economical sources of these natural additives and away from synthetic production has encouraged research into alternative supplies of these valuable compounds. Solid-state fermentation processes support the natural production of secondary metabolites, which represents most flavour and aroma compounds, while agro-industrial by-products are a low-value waste stream with a high potential for adding value. Accordingly, four filamentous fungi species with a history of use in the production of fermented foods and food additives were tested to ferment nine different agro-industrial by-products. Hundreds of volatile compounds were produced and identified using headspace (HS) solid-phase microextraction (SPME) coupled to gas chromatography–mass spectrometry (GC–MS). Four compounds of interest, phenylacetaldehyde, methyl benzoate, 1-octen-3-ol, and phenylethyl alcohol, were extracted and quantified. Preliminary yields were encouraging compared to traditional sources. This, combined with the low-cost substrates and the high-value natural flavours and aromas produced, presents a compelling case for further optimisation of the process.
... Vanillate hydroxylase activity has been observed in many ascomycetes and basidiomycetes and is suggested to play a major role in the degradation of the lignin G-units [31][32][33][34][35]. The identification of vhyA is an important finding which can unlock new strategies to engineer efficient fungal cell factories. ...
... However, deletion of hqdA did not result in a phenotype on ferulic acid nor did HqdA show activity on methoxyhydroquinone [39]. This suggests that methoxyhydroquinone can be converted to hydroxyquinol, which was also proposed for Paecilomyces variotii and S. pulverulentum [35,46]. However, the deletion of mhdA resulted in severely reduced growth on vanillin and vanillic acid and indicates that the conversion of methoxyhydroquinone to hydroxyquinol plays a minor role. ...
Background
The aromatic compounds vanillin and vanillic acid are important fragrances used in the food, beverage, cosmetic and pharmaceutical industries. Currently, most aromatic compounds used in products are chemically synthesized, while only a small percentage is extracted from natural sources. The metabolism of vanillin and vanillic acid has been studied for decades in microorganisms and many studies have been conducted that showed that both can be produced from ferulic acid using bacteria. In contrast, the degradation of vanillin and vanillic acid by fungi is poorly studied and no genes involved in this metabolic pathway have been identified. In this study, we aimed to clarify this metabolic pathway in Aspergillus niger and identify the genes involved.
Results
Using whole-genome transcriptome data, four genes involved in vanillin and vanillic acid metabolism were identified. These include vanillin dehydrogenase (vdhA), vanillic acid hydroxylase (vhyA), and two genes encoding novel enzymes, which function as methoxyhydroquinone 1,2-dioxygenase (mhdA) and 4-oxo-monomethyl adipate esterase (omeA). Deletion of these genes in A. niger confirmed their role in aromatic metabolism and the enzymatic activities of these enzymes were verified. In addition, we demonstrated that mhdA and vhyA deletion mutants can be used as fungal cell factories for the accumulation of vanillic acid and methoxyhydroquinone from guaiacyl lignin units and related aromatic compounds.
Conclusions
This study provides new insights into the fungal aromatic metabolic pathways involved in the degradation of guaiacyl units and related aromatic compounds. The identification of the involved genes unlocks new potential for engineering aromatic compound-producing fungal cell factories.
... On the other hand, it was reported that filamentous fungi may catabolize Van via different pathways: (1) non-oxidative decarboxylation to guaiacol, (2) oxidation of Van to protocatechuate which is followed by aromatic ring opening, and (3) oxidative decarboxylation to methoxy-p-hydroquinone (Lubbers et al., 2019). The non-oxidative degradation route of Van was described in a limited number of ascomycetes species such as Sporotrichum thermophile (Topakas et al., 2003), P. variotii (Rahouti et al., 1989), some Aspergillus species and yeasts (Guiraud et al., 1992;Huang et al., 1993). Guaiacol did not appear in HPLC chromatograms of Van degradation by B. nivea (Fig. 6). ...
Physicochemical pretreatments used to improve the bioconversion of recalcitrant lignocellulosic biomass generate toxic by-products, such as furan and phenols. In this study, 40 fungal strains were analyzed for their capability to grow with different concentrations of furfural, vanillin, 4-hydroxybenzaldehyde, and syringaldehyde. Byssochlamys nivea MUT 6321 showed promising growth performance when the inhibitors were used as single molecules and it was the only fungus that could grow when the four molecules were simultaneously present in the culture media. Further trials demonstrated that B. nivea was able to completely degrade furfural in 24 h and the phenolic aldehydes, as single molecules, in less than 11 days. In the presence of the three phenolic aldehydes, the fungus was able to transform them. However, when furfural was present in the mix, faster and preferential consumption of furfural instead of phenolic aldehydes was observed. This study provides important information for the use of this fungus to remove toxic compounds present in pretreated lignocellulosic biomass that could potentially lead to the enhancement of the efficiency of biofuels and chemicals production.
... Direct transformation of ferulic acid to vanillin by elimination of the side chain carbons may occur [31]. Vanillic acid is then formed by oxidation of vanillin [32]. Although vanillic acid was not found in suspension cell culture or culture filtrates incubated with LMC, this compound may be rapidly converted to other products. ...
Peroxidase secretion and activity in the oxidation of polyphenols bisphenol A (BPA, 2,2-bis(4-hydroxyphenyl)propane) and lignin model compound (LMC, guaiacylglycerol-β-guaiacylether) were observed in a suspension cell culture of liverwort Heteroscyphus planus. When BPA was co-incubated in a suspension cell culture of liverwort for 5 days, it was depleted by approximately 63%. Oxidation of BPA was observed in culture filtrates of liverwort, and most of the oxidation products were insoluble higher molecular-weight compounds (30%). The oxidative degradation products of BPA and LMC were analyzed by GC-MS and were identified by comparing their retention time and MS spectra with those of the authentic compounds. BPA was degraded to 4-isopropenylphenol and p-hydroxyacetophenone. The formation of these products was examined using [2H|4]-BPA. The lignin model compound was degraded to guaiacol, vanillin, coniferyl alcohol and ferulic acid. Biphenyl dehydrodimer was detected in both the reaction mixtures of the suspension cell culture and the culture filtrates incubated with the LMC. The dimer was identified as l,144,4'-dihydroxy-3,3'-dimethoxy-5,5'-biphenylene)- bis[8-(2"- methoxyphenoxy)-7,9- propanediol] by ID and 21) NMR analysis. The activity of secreted peroxidase in the suspension cell culture (0.045 U/mL) was slightly enhanced by addition of LMC (0.059 U/mL), p-cresol (0.064 U/mL), and 2,6-dimethoxyphenol (0.082 U/mL) 7 days after the beginning of incubation.
... 18 Therefore, white-rot fungi should be explored for their untapped potential in degradation of aromatic pollutants. Several non-white rot fungal strains such as Paecilomyces variotii 19 and Fusarium solani 20 can transform ferulic acid to vanillin along with several other metabolites over a period of 24 and 48 h of incubation, respectively. Although several non-white rot fungal strains were reported for bioconversion of ferulic acid, a limited number of white-rot fungi have been reported for degradation of ferulic acid. ...
... Although several non-white rot fungal strains were reported for bioconversion of ferulic acid, a limited number of white-rot fungi have been reported for degradation of ferulic acid. 19 Schizophyllum commune, a white-rot fungi, has degraded ferulic acid (initial concentration of 97.09 mg L −1 in 16 days of incubation) into 4-vinylguaiacol, which was further oxidized to vanillin and vanillic acid. 21 In another investigation, trametes sp. ...
Biodegradation of ferulic acid, by two white-rot fungal strains (Trametes hirsuta MTCC-1171 and Phanerochaete chrysosporium NCIM-1106) was investigated in this study. Both strains could use ferulic acid as a sole carbon source when provided with basal mineral salt medium. T. hirsuta achieved complete degradation of ferulic acid (350 mg L⁻¹) in 20 h, whereas P. chrysosporium degraded it (250 mg L⁻¹) in 28 h. The metabolites produced during degradation were distinguished by gas chromatography-mass spectrometry. Bioconversion of ferulic acid to vanillin by P. chrysosporium was also investigated. The optimum experimental conditions for bioconversion to vanillin can be summarized as follows: ferulic acid concentration 250 mg L⁻¹, temperature 35 °C, initial pH 5.0, mycelial inoculum 0.32 ± 0.01 g L⁻¹ dry weight, and shaking speed 150 rpm. At optimized conditions, the maximum molar yield obtained was 3.4 ± 0.1%, after 20 h of bioconversion. Considering that the degradation of ferulic acid was determined by laccase and lignin peroxidase to some extent, the possible role of ligninolytic enzymes in overall bioconversion process was also studied. These results illustrate that both strains have the potential of utilizing ferulic acid as a sole carbon source. Moreover, P. chrysosporium can also be explored for its ability to transform ferulic acid into value-added products.
... Gas chromatography (GC) was carried out on Thermo Scientific GCMS model ISQ at the Regional Center for Mycology and Biotechnology (RCMB), Al-Azhar University, Nasr City, Cairo, using the method described by Rahouti, et al. (1989). Thermo scientific (Trace 1310) column (TG-SQC, 15 m by 0.25mm) the temperature of injection was 250 ˚C. the column was heated at 100 ˚C for 1 min and then programmed to rise 10˚C/min to 280˚C. the carrier gas was He at flow rate of 1.5 ml/min. ...
... Furthermore, the cascade synthesis of vanillin from ferulic acid via 4-vinylguaiacol was also achieved by the immobilized phenolic acid decarboxylase and Cso2 [21]. In fact, many organisms such as Paecilomyces variotii, Pestalotia palmarum [22], Bacillus coagulans [23] and Enterobacter sp. Px6-4 [24] had been reported to metabolize ferulic acid to vanillin via 4-vinylguaiacol. ...
A 9-cis-epoxycarotenoid dioxygenase gene from Serratia sp. ATCC 39,006 (SeNCED) was overexpressed in soluble form in E.coli. SeNCED showed the maximum activity at 30 °C and pH 8.0, and it was stable relatively at range of pH 5–10 and temperature of 20 °C to 30 °C. SeNCED effectively catalyzes the side chain double bond cleavage of isoeugenol and 4-vinylguaiacol to vanillin. The kinetic constant Km values toward isoeugenol and 4-vinylguaiacol were 18.92 mM and 6.31 mM and Vmax values were 50.73 IU/g and 4.77 IU/g, respectively. Moreover, the SeNCED exhibited an excellent organic solvent tolerance and the enzyme activity was substantially improved at presence of 10% of trichloromethane. The produced vanillin was achieved at an around 0.53 g/L (3.47 mM) and 0.33 g/L (2.17 mM) after 8 h reaction at 4 mM of isoeugenol and 4-vinylguaiacol, respectively, using transformed Escherichia coli cells harboring SeNCED in the presence of trichloromethane.
... Se colocaran en cajas petri estériles sobre una cama de algodón humedecida con agua destilada estéril como soporte, cubierta por un papel filtro estéril; las semillas de Z. mays como control absoluto: se irrigaron con agua destilada y las semillas Z. mays como control relativo se alimentaron con solución mineral. Todas las semillas de control absoluto, control relativo y las tratadas con los transformados por R. etli se colocaron en obscuridad por 3 días, al germinar se dejaron en solárium otros 4 días, luego se midió: fenología: altura de planta, longitud de raíz; biomasa: peso fresco aéreo, peso fresco radical, luego se secó la parte aérea y radical (80C°/24h) en horno, para obtener el peso seco aéreo y radical; los datos experimentales se analizarón por Tukey con un nivel de significancia α de 0.05 mediante el programa Statgraphics Centurion 16.103 ® (Nishida & Fukuzumi, 1978;Rahouti et al., 1989). ...
Triticum aestivum (trigo) es un cultivo de alta demanda en consecuencia elevados niveles de paja se generan los que son normalmente incinerados con la consecuente generacion de gases efecto invernadero una oopcion de disponsicion inteligente es tratarla para obtener lignina residual de paja de trigo (LIREPATO) la que se degrada por hongos filtamentosos en aromaticos, lo que a su vez pueden ser convertido en sustancias promotoras de crecimiento vegetal (SUPROCEVE) por generos de bacterias promotoras de crecimiento vegetal,l (BAPROCEVE) Los resultados de esta investigacion demuestran que la degradacion de la LIREPATO en aromaticos por hongos y de los aromaticos en SUOROCEVE por generos de BAPROCEVE
... Por ejemplo con Trametes versicolor a partir de ácido ferúlico se generó alcohol veratrílico y veratrilaldehido (23), mientras que con Pycnoporus cinnabarinus se liberó vainillina con un rendimiento de 0,064 mg mL -1 en 7 días (9). Por otro lado, con el HML Paecilomyces variotii de ácido ferúlico se produjo ácido vainillínico, vainillina, alcohol vainillínico y 4 vinil-guayacol (24). ...
Background: Wheat straw is an agricultural waste, which contains 17% of lignin, a recalcitrant polymer with biotechnological potential provided it is depolymerized. Lignin depolymerization has attracted interest because it yields aromatics of industrial interest; chemical and physical methods are available but entail economic and environmental constraints. An alternative is to exploit the ligninolytic capacity of mitosporic fungi, such as Aspergillus and Penicillium spp. There are few reports on the use of these funguses in the generation of aromatics by lignin depolymerization. Objetives: To use Aspergillus and Penicillium spp in the biological generation of aromatics from semipurified residual wheat straw lignin. Methods: Funguses were grown in semipurified residual wheat straw lignin for 28 days; produced aromatics were followed using gas chromatography. Results: Obtained results indicate a range of aromatics produced, i.e. 3,5 mg mL-1 guaiacol, 3,3 vanillin, 3,2 hydroxybenzoic acid, 3,3 vanillinic, 10,1 syringic and 21,9 ferulic. Conclusions: Aspergillus and Penicillium represent an ecological option in the exploit of semi-purified residual lignin from wheat straw to generate aromatics in a shorter period from an abundant and cheap residue.
