Decarboxylation of Ferulic Acid to 4-Vinyl Guaiacol by <Emphasis Type="Italic">Streptomyces setonii</Emphasis>

4-Vinyl guaiacol (3-methoxy 4-hydroxystyrene) can be obtained by decarboxylation of ferulic acid by the strain Streptomyces setonii ATCC 39116. The formation of this metabolite was favoured by microaerobic conditions and the culture medium employed, increasing progressively the product concentration from 543.3 up to 885.1 mg/l when aeration level was diminished, reaching a highest volumetric productivity of 70.4 mg/l h and a product yield of 1.11 mol/mol. The identity of the metabolite was confirmed by gas chromatography–mass spectrometry. A metabolic study of ferulic acid and the main degradation products (ferulic acid, 4-vinyl guaiacol, protocatechuic acid, vanillyl alcohol, vanillic acid and vanillin) suggested that ferulic acid was the only substrate capable to be transformed into 4-vinyl guaiacol by this strain of S. setonii.     

Syringic acid metabolism by some white-rot,soft-rot,and brown-rot fungi

Syringic acid metabolism by four white-rot, two soft-rot, and two brown-rot fungi has been studied. The pathways for syringic acid metabolism have been studied in detail for the white-rot fungus,Sporotrichum pulverulentum, and a reaction sequence proposed. According to identified metabolites, the following reactions occur: reduction of the carboxyl group, hydroxylation and simultaneous decarboxylation, demethylation, and methylation of thep-hydroxyl group. In the case of the two soft-rot fungi,Petriellidium boydii andPhialophora mutabilis, rapid metabolism of syringic acid was observed, and the medium was depleted of the acid within 12 h. The formation of trimethoxybenzoic acid indicates an ability within the soft-rot fungi also to methylate the p-hydroxylic group. The two brown-rot fungi,Daedalea quercina andFomes pinicola, were poor metabolizers of syringic acid. However, demethylation was observed withF. pinicola and an unidentified product appeared in the culture solution ofD. quercina.     

Density functional theory study of the mechanism of the acid-catalyzed decarboxylation of pyrrole-2-carboxylic acid and mesitoic acid

The mechanisms of the acid-catalyzed decarboxylation of pyrrole-2-carboxylic acid and mesitoic acid have been investigated based on density functional theory calculations at the B3LYP/6-311G (d,p) level. A polarizable continuum model (PCM) has been established in order to evaluate the effects of solvents on these reactions. The results of the calculations indicate that the first step of the acid-catalyzed decarboxylation of the pyrrole-2-carboxylic acid has two possible pathways,that is,the proton of H3O+ a...     

Improved synthesis of 3-methoxy-4-hydroxymandelic acid by glyoxalic acid method

The most important industrial process for the synthesis of vanillin is performed in two steps involving an condensation reaction of glyoxalic acid with guaiacol followed by an oxidative decarboxylation of the intermediary 3-methoxy-4-hydroxymandelic acid (MHPA) formed, thereby producing not only vanillin, but also byproducts, which have to be eliminated. In the present study, we focused our efforts on the first step of vanillin synthesis, namely, the condensation reaction governing the yield of vanillin. The factors influencing the stability of glyoxalic acid were preliminarily evaluated to provide significant referential value for suppressing the dismutation of glyoxalic acid in the condensation reaction, and the results indicate that the stability of glyoxalic acid largely depends on the pH, temperature, and holding time of the feed solution. Then the process of the condensation reaction was optimized under the factors affecting the stability of glyoxalic acid, as well as the molar ratio of guaiacol to glyoxalic acid. Under the optimized conditions, the maximum yield of the condensation reaction can reach to 88%.     

Rapid Nanomolar Detection of Guaiacol from its Precursors Using a Core-shell Reversed-phase Column Coupled with a Boron-doped Diamond Electrode

The electrooxidation mechanisms of guaiacol (2-methoxyphenol), a food and beverage spoilage metabolite, and its precursors; vanillic acid (VA), vanillin, and ferulic acid (FA) were investigated by cyclic voltammetry (CV) with a boron-doped diamond (BDD) electrode. Reversed-phase liquid chromatography (RPLC) together with a BDD electrode poised at +1.6 V vs. Pd/H₂, was optimized for their sensitive detection. The separation was achieved in 60 s with a core-shell column (HALO C₁₈). The detection limits of these analytes ranged from 10–30 nM. The method was applicable for the analysis of guaiacol and its precursors from a popular commercial drink.     

Influence of culture conditions on lipopeptide production by Bacillus subtilis

Bacillus subtilis produces various families of lipopeptides with different homologous compounds. To produce “new molecules” with improved activities and to select strains that produced a reduced number of homologs or isomers, we studied the effects of different media on the nature of the synthesis of fatty acid chains for each lipopeptide family. This study focused on two B. subtilis strains cultivated in flasks. Optimized medium for lipopeptide production and Landymedium modified by replacing glutamic acid with other α-amino acids were used. We found that the intensity of production of homologous compounds depends on the strain and the culture medium. Analysis of these lipopeptides by high-performance liquid chromatography showed that the strain B. subtilis NT02 yielded various homologous compounds when cultivated in Landy medium (L-Glu), but primarily one homologous product in high relative amounts when cultivated in the optimized medium. Mass spectrometric analysis and determination of the amino acid composition of this molecule enabled us to identify it as Bacillomycine L c15.     

Growth of lignocellulosic-fermenting fungi on different substrates under low oxygenation conditions

Four soil fungi able to grow under low oxygenation conditions were selected and used in studies to determine the production of enzymes that promote the degradation of lignocellulosic materials. The capacity of these fungi to ferment such materials was also investigated. The fungi were grown in sugarcane bagasse and sawdust at final concentrations of 4 and 10%, as the carbon sources. The strains were cultivated under microaerophilic and combined conditions of oxygenation (aerobic followed by microaerophilic conditions). The results obtained with the basidiomycete specie, Trichocladium canadense, Geotrichum sp., and Fusarium sp. suggest that they prefer lower oxygen concentration for growth and enzyme production. Lignocellulolytic activities were detected in all strains but varied with the carbon source used. The highest levels of these activities were produced by the Basidiomycete specie and Fusarium sp. Ethanol and other nongaseous fermentation products were detected following high-performance liquid chromatography analysis using a supelcogel C-610H column, demonstrating the fermentative capability of these strains. In view of their ability to produce enzymes necessary for the breakdown of lignocellulosic materials and to utilize most of the degradation products for growth, these strains have a great potential for biotechnological application.     

Renewable benzoxazine monomer from Vanillin: Synthesis,characterization, and studies on curing behavior

A novel renewable based benzoxazine, 3‐(furan‐2‐ylmethyl)?8‐methoxy‐3,4‐dihydro‐2H‐1,3‐benzoxazine‐6‐formyl (Va‐Bz), has been synthesized from a lignin derived chemical “vanillin” without solvents. Poly (Va‐Bz) has high Tg and excellent thermal and adhesive properties. A mechanism of cross‐linking, due to electrophilic substitution at furan and decarboxylation of carboxylic group of benzene ring, is suggested. © 2013 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2014 , 52, 7–11     

Proteomics of edible mushrooms: A mini‐review

Mushrooms are considered an important food for their traditionally famous nutritional and medicinal values, although much information about their potential at the molecular level is unfortunately unknown. Edible mushrooms include fungi that are either collected wild or cultivated. Many important species are difficult to cultivate but attempts have been made with varying degrees of success, with the results showing unsatisfactory economical cultivation methods. Recently, proteomic analysis has been developed as a powerful tool to study the protein content of fungi, particularly basidiomycetes. This mini‐review article highlights the contribution of proteomics platforms to the study of edible mushrooms, focusing on the molecular mechanisms involved in developmental stages. This includes extracellular and cytoplasmic effector proteins that have potential or are involved in the synthesis of anticancer, antidiabetic, antioxidant, and antibiotic, in blood pressure control, in the supply of vitamins and minerals, and in other responses to environmental changes. The contribution of different proteomics techniques including classical and more advanced techniques is also highlighted.     

Heterotrophic Nitrifying and Oxygen Tolerant Denitrifying Bacteria from Greenwater System of Coastal Aquaculture

In this work, herbivorous fish Mugil cephalus has been cultured to secrete protein rich green slime, which helps nitrifying and oxygen tolerant denitrifying bacteria to grow and colonize. Four strains representing Alcaligenaceae family have been isolated from greenwater system and characterized using biochemical test, fatty acid methyl ester (GC-FAME) analysis, 16S rRNA and functional gene approaches. They were tested for an ability to nitrify ammonia and nitrite aerobically. Two strains showed notable nitrification activity, when grown in a mineral salts medium containing ammonium sulfate and potassium nitrite. Functional gene analysis confirmed the presence of nitrous oxide reductase (nosZ) gene showing that they have an oxygen-tolerant denitrification system. It has been proposed that Alcaligenes faecalis strains heterotrophically nitrify ammonia into nitrite via formation of hydroxyl amine, which is oxidized to nitrous oxide using oxygen or nitrite as electron acceptor. These results provide a possible advantage of having nitrification and denitrification capabilities in the same organism, which plays an important role in biological wastewater system.     

Overtone-induced decarboxylation: a potential sink for atmospheric diacids

Atmospheric photochemistry induced by solar excitation of vibrational overtone transitions has recently been demonstrated to be of importance in cleaving weak bonds (in HO(2)NO(2)) and inducing intramolecular rearrangement followed by reaction (in H(2)SO(4)). Here, we propose another potentially important process: the decarboxylation of organic acids. To demonstrate this possibility, we have calculated the decarboxylation pathways for malonic acid and its monohydrate. The barrier to the gas-phase decarboxylation was calculated to be in the range 26-28 kcal/mol at the B3LYP/6-311++G(3df,3pd) level of theory, in good agreement with previous results. The transition state is a six-membered ring structure which is accessed via concerted O-H and C-C stretches; excitation of v(OH) > or = 3 of either one of the OH stretching modes is sufficient to supply the energy needed for the decarboxylation. A low-energy isomer of the malonic acid-water complex forms an eight-membered, multiply hydrogen bonded structure, bound by 3-6 kcal/mol, somewhat less stable than the lowest energy, six-membered ring isomer. Decarboxylation of such complexes uses water as a catalyst; the water accepts an acidic proton from one malonic acid group and transfers a proton to the carbonyl of the other acid group. The barrier for this process is 20-22 kcal/mol, suggesting that complexes excited to v(OH) > or = 2 possess sufficient energy to react. Using estimated absorption cross sections for the OH overtone transitions, we suggest that the overtone-induced decarboxylation of malonic acid and its water complex is competitive with wet deposition of the acid and with gas-phase reaction with OH for removal of the acid.     

A biotechnological process involving filamentous fungi to produce natural crystalline vanillin from maize bran

A new process involving the filamentous fungi Aspergillus niger and Pycnoporus cinnabarinus has been designed for the release of ferulic acid by enzymic degradation of a cheap and natural agricultural byproduct (autoclaved maize bran) and its biotransformation into vanillic acid and/or vanillin with a limited number of steps. On the one hand, the potentialities of A. niger I-1472 to produce high levels of polysaccharide-degrading enzymes including feruloyl esterases and to transform ferulic acid into vanillic acid were successfully combined for the release of free ferulic acid from autoclaved maize bran. Then vanillic acid was recovered and efficiently transformed into vanillin by P. cinnabarinus MUCL 39533, since 767 mg/L of biotechnologic vanillin could be produced in the presence of cellobiose and XAD-2 resin. On the other hand, 3-d-old high-density cultures of P. cinnabarinus MUCL39533 could be fed with the autoclaved fraction of maize bran as a ferulic acid source and a. niger I-1472 culture filtrate as an extracellular enzyme source. Under these conditions, P. cinnabarinus MUCL39533 was shown to directly biotransform free ferulic acid released from the autoclaved maize bran by A. niger I-1472 enzymes into 584 mg/L of vanillin. These processes, involving physical, enzymic, and fungal treatments, permitted us to produce crystallin vanillin from autoclaved maize bran without any purification step.     

Andrographolide: Solving Chemical Instability and Poor Solubility by Means of Cocrystals

The bioactive agent andrographolide was screened with pharmaceutically acceptable coformers to discover a novel solid form that will solve the chemical instability and poor solubility problems of this herbal medicine. Liquid‐assisted grinding of andrographolide with GRAS (generally regarded as safe) coformers in a fixed stoichiometry resulted in cocrystals with vanillin (1:1), vanillic acid (1:1), salicylic acid (1:1), resorcinol (1:1), and guaiacol (1:1). All the crystalline products were characterized by thermal, spectroscopic, and diffraction methods. Interestingly, even though the cocrystals are isostructural, their physicochemical properties are quite different. The andrographolide–salicylic acid cocrystal completely inhibited the chemical transformation of andrographolide to its inactive sulfate metabolite, and moreover, the cocrystal exhibited a dissolution rate that was three times faster and a drug release that was two times higher than pure andrographolide.     

Lignin degrading ability of selected aspergillus Spp.

Most lignin research has been on wood-rot fungi and not on other lignolytic organisms. Members of the genusAspergillus inhabit lignin-rich environments, and we have studied their relative lignin-degrading potential.Aspergillus fumigatus, A. japonicus, A. niger, andA. terreus were tested for their ability to metabolize¹⁴C-labeled aromatic compounds. The species tested decarboxylated, demethoxylated, and cleaved the rings of coumaric, ferulic, vanillic, veratric, and anisic acids. More than 90% of C-ring-labeled ferulic and vanillic acids disappeared from the medium in 96 h of cultivation. More than half of the above was respired, the rest was incorporated in unknown form into the mycelium. Mycelia were homogenized and about 3% of the initial label was found in TCA precipitate of the cell-free supernatant. Protocatechuic acid 3,4-dioxygenase (EC 1.13.11.3) and catechol 1,2-dioxygenase (EC 1.13.11.1) activities were detected in the mycelial extracts of theAspergillus spp. All theAspergillus spp. were capable of degrading both aromatic and carbohydrate components of water-soluble lignocarbohydrate complexes (LCC) from wheat straw. The degradation of the aromatic moiety of soluble LCC with apparent molecular mass more than 100,000 daltons was far more active in theAspergillus spp. than in the whiterot fungi tested; i.e.Polyporus versicolor, Pleurotus ostreatus, andForties annosus. The aromatics present in the soluble LCC, as well as a variety of lignin-related compounds tested, did not affect the production of hemicellulases byA. japonicus. Aspergillus spp. degraded¹⁴C-dehydrogenative polymerizates, converting carbon from the ring as well as from the -O¹⁴CH₃ groups to¹⁴CCO₂.¹⁴CO₂ release after 21 d did not exceed 10% of the total¹⁴C input. This situation is comparable to some white-rot fungi. Lignosulfonate was poorly degraded byA. japonicus, but clearly modified.Fomes annosus was able to grow much better on lignosulfonate whenA. japonicus had previously grown on it.Aspergillus spp. grew efficiently on wheat straw, utilizing lignin and some carbohydrates, and rendering the remaining carbohydrates more available to attack of carbohydrases.     

Acrylamide formation in food: a mechanistic perspective

Earliest reports on the origin of acrylamide in food have confirmed asparagine as the main amino acid responsible for its formation. Available evidence suggests that sugars and other carbonyl compounds play a specific role in the decarboxylation process of asparagine, a necessary step in the generation of acrylamide. It has been proposed that Schiff base intermediate formed between asparagine and the sugar provides a low energy alternative to the decarboxylation from the intact Amadori product through generation and decomposition of oxazolidin-5-one intermediate, leading to the formation of a relatively stable azomethine ylide. Literature data indicate the propensity of such protonated ylides to undergo irreversible 1,2-prototropic shift and produce, in this case, decarboxylated Schiff bases which can easily rearrange into corresponding Amadori products. Decarboxylated Amadori products can either undergo the well known beta-elimination process initiated by the sugar moiety to produce 3-aminopropanamide and 1-deoxyglucosone or undergo 1,2-elimination initiated by the amino acid moiety to directly generate acrylamide. On the other hand, the Schiff intermediate can either hydrolyze and release 3-aminopropanamide or similarly undergo amino acid initiated 1,2-elimination to directly form acrylamide. Other thermolytic pathways to acrylamide--considered marginal at this stage--via the Strecker aldehyde, acrolein, and acrylic acid, are also addressed. Despite significant progress in the understanding of the mechanistic aspects of acrylamide formation, concrete evidence for the role of the different proposed intermediates in foods is still lacking.     

Mechanistic study of palladium‐catalyzed decarboxylative coupling of phenylpropiolic acid and aryl iodide

The mechanism of decarboxylative coupling of phenylpropiolic acid and methyl 4‐iodobenzoate was investigated by gas chromatographic analysis and monitoring the decarboxylation reaction by FT‐IR spectroscopy. Among many possible pathways, we found that the mechanism involves two possible pathways, namely the decarboxylation of phenylpropiolic acid and that proceeding through the carboxylated palladium complex. Copyright © 2012 John Wiley & Sons, Ltd.     

The influence of modified EK2010 catalyst to the decarboxylation of oleic acid

It is known that the zeolite type catalytic system is the main catalyst of decarboxylation process. However, the influence of modification of acid–base properties of this type of catalyst to the decarboxylation reaction of organic acids is not investigated widely. Therefore, the modification of the bifunctional “EK2010” catalyst with MgO is conducted in the submitted paper. The influence of this modification to the route of decarboxylation process of organic acids and yield of target products have been investigated in the example of oleic acid. It is found out that, when the basicity is increasing, the modification occurs on the surface of catalyst and the activity of the catalyst is rising and decarboxylation reaction of oleic acid is going very well. The catalysts was investigated with XRD, XRF, TGA and IR techniques.     

营养缺陷型酿酒酵母AY生长代谢的热动力学研究

用微量热法研究了尿嘧啶缺陷型酿酒酵母AY,和该菌株分别经穿梭质粒pYLZ-2、重组质粒pYLZ-2/f27、pYLZ-2/622转化的3种菌株的生长代谢热动力学.尿嘧啶缺陷型酿酒酵母AY在基本培养基中不生长,没有代谢热效应产生,而在葡萄糖蛋白胨培养基和丰富营养培养基(YPD)中生长,并且在YPD中生长最好;向基本培养基中加入尿嘧啶后,AY能够生长,而且随着尿嘧啶浓度的增加,其生长速率常数增大;经质粒转化的3个菌株均能在基本培养基中生长,代谢产热曲线各不相同,与转入质粒的结构、功能密切相关.研究结果表明了各菌株遗传特征的差异.     

Black Liquor Decolorization by Selected White-Rot Fungi

Five different strains of white-rot fungi have been tested for their ability to decolorize black liquor on plates and on solid-state fermentation using vermiculite as the solid inert support. Since the high salt concentration inhibited the growth of all fungi, the black liquor was dialyzed against distilled water prior to use. A preliminary step on plates was carried out to qualitatively determine the capacity of the fungal strains for black liquor decolorization. Out of the five fungi studied, Phanerochaete sordida, Pycnoporus sanguineus, and Trametes elegans exhibited the more conspicuous decolorization halos in malt extract medium, while the decolorization by all the strains was not evident when a defined culture medium was used. Cultures on solid-state fermentation using vermiculite as solid support were also tested, the liquid phase was malt extract or glucose-based medium and supplemented with different black liquor concentrations. Decolorization of black liquor was largely affected by the fungal strain, the concentration of black liquor, and the carbon source. The percentage of color removal ranged from 6.14% to 91.86% depending on the fungal strain and culture conditions. Maximal decolorization was observed in malt extract cultures after 60 cultivation days. Interestingly, decolorization in malt extract medium increased with increasing black liquor concentration. The highest decolorization value was achieved by Steccherinum sp. which reduced up to 91.86% the color of the black liquor in malt extract medium; this percentage is equivalent to 5.2 g L⁻¹ of decolorized black liquor, the highest value reported to date. Traditional technologies used for the treatment of black liquor are not always effective and may not to be an environmentally friendly process. Vermiculite–white-rot fungi systems are presented in this work as a promising efficient alternative for the treatment of black liquor.