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1.
The accumulation of phenolic acids in soil is one of the main problems associated with continuous cropping of peanut. Although laccases secreted by fungi can efficiently transform phenolic acids, there are few reports on the use of these enzymes to bioremediate continuous cropping soil. Food waste and wheat straw are waste products; however, they could be used productively as resources for laccase production by the endophytic fungus Phomopsis liquidambari B3. We cultured Phomopsis liquidambari B3 in medium containing food waste as the main nitrogen source and wheat straw as the main carbon source. In order to study the effects of fermentation liquid on phenolic acid degradation, rhizosphere soil microbial communities and peanut seedling growth, the fermentation product, which had high laccase activity, was added to continuously cropped soil of peanut. The concentration of 4-hydroxybenzoic acid, vanillic acid, and coumaric acid in soil had decreased by 57.4, 52.5, and 49.4%, respectively, compared with no-treatment control during 28 days. Analysis of denaturing gradient gel electrophoresis profiles showed that the bacterial and fungal community structures in rhizosphere soil were affected by changes in the phenolic acids concentration. The biomass of peanut plants and the number of root nodules were increased 68.3% and 5.9-fold, respectively. These results showed that the laccase product reduced the accumulation of phenolic acids in soil, the decrease in phenolic acids concentration and the increase in certain dominant microorganisms promoted peanut seedling growth and nodulation. This technology provides a new strategy for bioremediation of continuous cropping soil, while simultaneously reducing waste and protecting the environment.  相似文献   

2.
The decolorization and degradation of anionic sulphonated azo dye (Reactive orange 16 (RO16)), which is suspected to be carcinogenic, were investigated using ozone. The decolorization process of the reactive dye was carried out by bubbling ozone at the bottom of a bubble column reactor containing the dye solution. The effect of pH, reaction time, dye concentration, ozone concentration, and decolorization time was studied. Also, degradation products and possible degradation mechanism were investigated. The results showed that ozonation was a highly effective way to remove color from wastewater. The color of a synthetic waste solution containing water-soluble reactive dye was reduced to 69.69 % under the basic condition (pH 12), with complete RO16 degradation occurring in 8 min. Ozone consumption continued for a further 16 min after which time most of the degradation reactions were complete. Kinetic studies showed that direct ozonation of the aqueous dyes represented a pseudo-first-order reaction with respect to the dye. The apparent rate constant increased with both the applied ozone dose and higher pH values and declined logarithmically with the initial dye concentration. Intermediates such as 6-acetylamino-3-aminonaphthalene-2-sulfonic acid, 2-(4-nitrosophenyl) sulfonylethyl hydrogen sulfate, and 6-acetamido-4-hydroxy-3-nitroso naphthalene-2-sulfonic acid were detected by gas chromatograph coupled with mass spectrometry in the absence of pH buffer, while nitrate and sulfate ions and formic, acetic, and oxalic acids were detected by ion chromatography.  相似文献   

3.
In this study, in-house isolated laccase isoforms, i.e., Lac-I and Lac-II of the basidiomycete Pycnoporus sanguineus (CS43), were evaluated in relation to their Remazol Brilliant Blue R (RBBR) dye degradation capacity. A modified Dhouib medium additionally supplemented with 3% ethanol as a secondary inducer was used to propagate P. sanguineus CS43 for enhanced production of laccase under liquid state fermentation. The crude laccase extract was purified by passing through ion exchange diethylaminoethanol (DEAE)-Sepharose and gel filtration-based Sephadex G-200 column chromatography. The purified laccase fractions were subjected to the electrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed two laccase isoforms Lac-I and Lac-II with 66 and 68 kDa, respectively. To explore the industrial applicability, for RBBR dye, degradation efficiencies ranged from 82 to 88% after 3 h of incubation for both; Lac-I and Lac-II at both concentrations were recorded. However, with 8 U/mL, the degradation ranged between 70 to 80% during the first 5 min of incubation. Enhanced degradation of RBBR dye was obtained in the presence of violuric acid and N-hydroxypthalamide as laccase mediators. Finally, using RBBR as a substrate kinetic characterization of both Lac-I and Lac-II isoforms was performed that revealed K m (0.243 and 0.117 mM for Lac-I and Lac-II) and V max (1.233 and 1.012 mM/Sec for Lac-I and Lac-II) values, respectively.  相似文献   

4.
为了进一步掌握高效降解黄曲霉毒素B_1的食用菌SJ-1的特性,考察了温度、pH以及金属离子对食用菌SJ-1漆酶酶活的影响,同时研究了该漆酶降解黄曲霉毒素B_1的降解曲线和降解方程。结果表明,食用菌SJ-1漆酶的最适反应温度为35℃,最适pH值为3.0。Cu~(2+)对该漆酶酶活有一定的激活作用,而Fe~(2+)、Ca~(2+)、Mg~(2+)、Na~+、K~+对其酶活都有较强的抑制作用。食用菌SJ-1漆酶降解黄曲霉毒素B_1的方程为y=-0.1712x~(2+)20.107x-76.587,其中x为反应时间,范围为0~48 h,y为黄曲霉毒素B_1降解量(ng),该方程在0~48 h内,能够推算出食用菌SJ-1漆酶降解黄曲霉毒素B_1的降解量。本研究结果为食用菌漆酶降解黄曲霉毒素的应用提供了理论指导和技术支持。  相似文献   

5.
Elucidating the mechanism of laccase and tyrosinase in wheat bread making   总被引：2，自引：0，他引：2  
Cross-linking enzymes generate covalent bonds in and between food biopolymers. These enzymes are interesting tools for tailoring dough and bread structures, as the characteristics of the biopolymers significantly determine the viscoelastic and fracture properties of dough and bread. In this study, the influence of oxidative cross-linking enzymes, tyrosinase from the filamentous fungus Trichoderma reesei and laccase from the white rot fungus Trametes hirsuta, on dough and bread were examined. Oxidation of low molecular weight phenolic model compounds of flour, cross-linking of gluten proteins, dough rheology, and bread making were characterized during or after the enzymatic treatments. In the dough and bread experiments, laccase and tyrosinase were also studied in combination with xylanase. Of the model compounds tyrosine, p-coumaric acid, caffeic acid, ferulic acid, and Gly-Leu-Tyr tripeptide, tyrosinase oxidized all except ferulic acid. Laccase was able to oxidize each of the studied compounds. The phenolic acids were notably better substrates for laccase than l-tyrosine. When the ability of the enzymes to cross-link isolated gliadin and glutenin proteins was studied by the SDS-PAGE analysis, tyrosinase was found to cross-link the gliadin proteins effectively, whereas polymerization of the gliadins by laccase was observed only when a high enzyme dosage and prolonged incubation were used. Examination of large deformation rheology of dough showed that both laccase and tyrosinase made doughs harder and less extensible, and the effects increased as a function of the enzyme dosage. In bread making, interestingly, the pore size of the breads baked with tyrosinase turned out to be remarkably larger and more irregular when compared to that of the other breads. Nevertheless, both of the oxidative enzymes were found to soften the bread crumb and increase the volume of breads, and the best results were achieved in combination with xylanase.  相似文献   

6.
Caffeic acid o-quinone (CQ) was prepared by oxidation of caffeic acid with o-chloranil in organic media. The reaction between the purified CQ and cyanidin 3-glucoside (Cy 3-glc, o-diphenolic anthocyanin) was monitored by HPLC, and quantitative analyses were performed to establish the stoichiometry of the reaction. The results indicate that Cy 3-glc is degraded by a coupled oxidation mechanism with integration of CQ into the degradation products. The ratio of degraded Cy 3-glc to CQ incorporated into the condensation products was approximately 2.0. No brown products could be detected, only a slight orange color. Moreover, the addition of purified polyphenol oxidase to the slightly colored media resulted in the disappearance of the caffeic acid formed from the reaction of coupled oxidation (Cy 3-glc/CQ) and the formation of brown polymers. The degradation products were isolated by gel filtration on Sephadex G-25. The UV-vis spectra and chemical analysis (acidic hydrolysis) of the degradation products suggest that they resulted from the condensation of caffeic acid and Cy 3-glc. HPLC analysis showed that the partial purified fraction contained a mixture of complex condensation products.  相似文献   

7.
When vanillic acid was incubated with an extracellular laccase from the fungus Rhizoctonia praticola, the formation of various oligomeric products was observed ranging from dimers to pentamers. The three dimers isolated by thin-layer chromatography had molecular ions of 304, 304 and 334, respectively, which corresponded to two dimers (methoxy-p-benzoquinone-vanillic acid) with C-O (2-methoxy-6-(2'-methoxy-4'-carboxyphenoxy)-1,4-benzoquinone) and C-C (2-methoxy-6-(2'-hydroxy-3'-methoxy-5'-carboxyphenyl)-1,4-benzoquinone) coupling, and dehydrodivanillic acid (m/z 334) (2,2'-dihydroxy-3, 3'-dimethoxy-5,5'-dicarboxybiphenyl). The identity of these compounds and their methylated derivatives was confirmed by mass and NMR spectrometric analysis. The oxidative coupling of vanillic acid is also catalyzed by peroxidase but not by tyrosinase.  相似文献   

8.
A novel dye degrading bacterium capable of decolorizing and mineralizing four different dyes (Methyl red, Orange II, Direct red 80, and Direct blue 71) was isolated from textile industrial wastewater using the selective enrichment technique. The bacterium was identified as Pseudomonas aeruginosa. More than 80 % decolorization of Direct red 80 was obtained under microaerophilic conditions in 48 h, whereas only 10 % color removal was obtained under oxic conditions at the same time. Subsequent aeration of the decolorized medium resulted in the mineralization of the metabolic intermediates generated after azo bond cleavage by P. aeruginosa as confirmed by total organic carbon content and high-performance liquid chromatography analyses. The degradation products were characterized by Fourier transform infrared spectrometer and nuclear magnetic resonance techniques whereas the biotoxicity profile of the samples were evaluated using the brine shrimp lethality test assay. Data from this study provide evidence of dye mineralization and detoxification by a monoculture of P. aeruginosa in successive microaerophilic/oxic stages.  相似文献   

9.
Due to the production of lignocellulose-degrading enzymes, saprotrophic basidiomycetes can significantly contribute to the turnover of soil organic matter. The production of lignin- and polysaccharide-degrading enzymes and changes of the chemical composition of litter were studied with three isolates from a Quercus petraea forest. These isolates were capable of fresh litter degradation and were identified as Gymnopus sp., Hypholoma fasciculare and Rhodocollybia butyracea. Within 12 weeks of incubation, H. fasciculare decomposed 23%, R. butyracea 32% and Gymnopus sp. 38% of the substrate dry mass. All fungi produced laccase and Mn-peroxidase (MnP) and none of them produced lignin peroxidase or other Mn-independent peroxidases. There was a clear distinction in the enzyme production pattern between R. butyracea or H. fasciculare compared to Gymnopus sp. The two former species caused the fastest mass loss during the initial phase of litter degradation, accompanied by the temporary production of laccase (and MnP in H. fasciculare) and also high production of hydrolytic enzymes that later decreased. In contrast, Gymnopus sp. showed a stable rate of litter mass loss over the whole incubation period with a later onset of ligninolytic enzyme production and a longer lasting production of both lignin and cellulose-degrading enzymes. The activity of endo-cleaving polysaccharide hydrolases in this fungus was relatively low but it produced the most cellobiose hydrolase. All fungi decreased the C/N ratio of the litter from 24 to 15-19 and Gymnopus sp. also caused a substantial decrease in the lignin content. Analytical pyrolysis mass spectrometry of litter decomposed by this fungus showed changes in the litter composition similar to those caused by white-rot fungi during wood decay. These changes were less pronounced in the case of H. fasciculare and R. butyracea. All fungi also changed the mean masses of humic acid and fulvic acid fractions isolated from degraded litter. The humic acid fraction after degradation by all three fungi contained more lignin and less carbohydrates. Compared to the decomposition by saprotrophic basidiomycetes, litter degradation in situ on the site of fungal isolation resulted in the relative enrichment of lignin and differences in lignin composition revealed by analytical pyrolysis. It can most probably be explained by the participation of non-basidiomycetous fungi and bacteria during natural litter decomposition.  相似文献   

10.
The impact of processing on nonenzymatic antioxidant degradation and lipid oxidation leading to off-flavor development in potato flakes during storage was investigated. Lipoxygenase activity measurements in parallel with the analysis of lipid oxidation products (oxylipins) profiles using HPLC showed that the processing conditions used inhibited efficiently enzymatic lipid oxidation. However, nonenzymatic lipid oxidation products were found throughout processing and in fresh potato flakes. Furthermore, these autoxidative processes cannot be inactivated by the main endogenous nonenzymatic antioxidants in potato tubers (ascorbic acid, phenolic compounds and carotenoids), as these antioxidants are degraded during processing. Indeed, leaching and thermal treatments taking place during processing lead to a decrease of about 95%, 82% and 27% in the content of ascorbic acid, phenolic compounds and carotenoids, respectively. Therefore, storage is a critical step to prevent off-flavor development in potato flakes. Specific attention has thus to be paid on the use of efficient exogenous antioxidants as well as on storage conditions.  相似文献   

11.
The decolorization of the recalcitrant dye Remazol Brilliant Blue R (RBBR) by the culture filtrate of Polyporus sp. S133 and the effect of various environmental factors were investigated. Both biodegradation and biosorption were playing an important role in bioremoval mechanisms. The highest biosorption of RBBR in Polyporus sp. S133 was shown by all carbon sources such as sucrose, glucose, fructose, and starch. No biosorption was shown by the addition of aromatic compounds and metal ions; 97.1?% RBBR decolorization was achieved in 120-rpm culture for 96?h, as compared to 49.5?% decolorization in stationary culture. Increasing the shaking rotation of the culture to more than 120?rpm was proven to give a negative effect on decolorization. The highest production of laccase was shown at pH 4 and constantly decreases when the pH level increases. The addition of glucose, ammonium tartrate, Cu2+, and protocatechuic acid was the suitable environmental condition for RBBR decolorization. There was a positive relationship between all environmental conditions and laccase production in the decolorization of RBBR.  相似文献   

12.
Degradation products of ultraviolet (UV-C, 254 nm) treated ascorbic acid (AA) are reported. Analysis by high-performance liquid chromatography-mass spectroscopy (HPLC-MS) conducted in a 0.5% malic acid model juice system (pH 3.3) demonstrated increased degradation of AA above untreated controls with concomitant increases in dehydroascorbic acid (DHA) and 2,3-diketogulonic acid (DKGA) levels. Electron spin resonance (ESR) spectroscopy studies, conducted in phosphate buffer (pH 7.0) to increase detection sensitivity, demonstrated that ascorbyl radical (AA?) formation occurs simultaneously with AA degradation. Consistent with a previous study in which UV treatments were shown to accelerate dark storage degradation, AA? radicals continued to form for up to 200 min after an initial UV treatment. Results from this study suggest that the mechanism for UV-induced degradation is the same as the general mechanism for metal-catalyzed oxidation of AA in juice.  相似文献   

13.
超低浓度马来酸水解玉米芯纤维素   总被引：1，自引：0，他引：1  
为考察超低浓度马来酸对玉米芯纤维素的水解性能,该文采用高温液态水预处理和超低马来酸水解相结合的两步法。3,5-二硝基水杨酸(DNS)比色法和高效液相色谱法(HPLC)分析表明,第一步预处理(200℃,10min,4MPa,500r/min,液固比20:1mL/g)玉米芯可获得12.24g/L还原糖,半纤维素转化率91.76%,损失3.61%的纤维素;其残渣进行第二步酸水解(质量分数0.1%,220℃,20min,4MPa,500r/min,液固比20:1mL/g)可获得9.94g/L还原糖,纤维素转化率达95.17%,约1/3转化为糖。气相色谱-质谱联用(GC-MS)分析表明,第二步水解液中含有多种木质素降解副产物,如苯酚、苯甲酸等,带有多种活泼基团,可能与糖降解物反应,加快葡萄糖降解正反应的进行。改进反应器,使得糖降解物和木质素降解物及时排出,可提升马来酸水解性能,为马来酸在生物质水解领域的应用提供参考。  相似文献   

14.
To study a way to covalently link arabinoxylans and proteins using a fungal laccase from the fungus Pycnoporus cinnabarinus, the effect of cysteinyl caffeic acid on the cross-linking of wheat arabinoxylans was investigated by means of capillary viscometry and RP-HPLC of alkali labile phenolic compounds. Cysteinyl caffeic acid provoked a delay in gelation and in the consumption of the esterified ferulic acid on arabinoxylans. When reacting free ferulic acid and cysteinyl caffeic acid with laccase, the ferulic acid consumption and the dehydrodimers production were also diminished. These results suggest that cysteinyl caffeic acid is oxidized while reducing the semiquinones of ferulic acid produced by laccase. Thus, ferulic acid could not be oxidized into dimers until all cysteinyl caffeic acid was consumed, preventing the cross-linking of feruloylated arabinoxylan chains. A similar mechanism is proposed in the case of caffeic acid and of L-Dopa.  相似文献   

15.
The effects of a laccase from the fungus Pycnoporus cinnabarinus on the mixing of a wheat flour dough with or without added ferulic acid (FA) were studied. Laccase reduced dough time‐to‐peak and accelerated dough breakdown in comparison with the control. Its effect was enhanced with FA. The water extractability of arabinoxylans (AX) increased during mixing of a dough free of added laccase, especially with exogenous FA. At the same time, the extractability of FA decreased during mixing. Added FA may have competed with endogenous AX feruloyl esters, inhibiting partly oxidative gelation. Laccase decreased AX extractability by chain cross‐linking through oxidative dimerization of feruloyl esters. FA and, moreover, FA plus laccase, increased the oxidation of sulfhydryl (SH) groups. FA and, even more, FA in combination with laccase, increased the rate of protein depolymerization during mixing. FA and the products of FA laccase oxidation participated in a redox reaction involving SH groups. A coupling reaction involving enzymatically generated feruloyl radicals and thiol radicals generated through the mechanical breakdown of inter‐chain disulfide bonds might explain these results.  相似文献   

16.
We studied the influence of an immobilized laccase from Trametes versicolor on non-extractable residue (NER) formation of the systemic fungicide 14C-metalaxyl in soil. We added the enzyme (130 mU/g DW) to soil sterilized by gamma irradiation and observed that the amount of NER (6.3 % of applied radioactivity) after 10 days of incubation was enhanced about twofold compared to the sterile soil without laccase addition. Residues formed within samples without enhanced enzyme activity were mainly bound via ester linkages to all fractions of humic matter, i.e., fulvic acids, humic acids, non-humines, and humines, respectively. In contrast, residues formed in presence of immobilized laccase were more strongly bound by covalent linkages such as ether and C-C bonds, especially to humic acids. After chemical degradation of the humic matter, it could be observed that all NER contained the first major transformation product, i.e., metalaxyl acid. The findings underline that the residue formation of metalaxyl in soil may be partly catalyzed by immobilized extracellular oxidative enzymes through oxidative coupling reactions within the humic matter.  相似文献   

17.
Polyporus sp. S133 fungi were selected based on their ability to degrade anthracene in liquid media. The degradation efficiency of anthracene increased by adding 0.5% Tween 80 to reach 71%; agitation at 120 rev/min increased the degradation to 92% after 30?days of incubation. Enzymes such as manganese peroxidase (MnP), lignin peroxidase (LiP), laccase, 1,2-dioxygenase and 2,3-dioxgenase were produced by Polyporus sp. S133 during incubation, and the highest enzyme activity was 182.3 U l?1 by 1,2-dioxygenase after 20?days of incubation. These results indicate that ligninolytic and dioxygenase enzymes secreted from Polyporus sp. S133 could play an important role in anthracene degradation efficiency. The metabolites detected through the degradation pathway were anthraquinone, phthalic acid, benzoic acid and catechol.  相似文献   

18.
低毒病毒是一类存在于板栗疫病菌细胞质中自主复制的无衣壳正链RNA病毒。感染低毒病毒后,板栗疫病菌的致病力显著降低,色素分泌减少,菌丝体由感染病毒前的桔黄色变为白色,产孢量降低或不产孢,漆酶表达水平明显下降。低毒病毒侵染性克隆的获得以及高效转化和转染体系的建立,使得低毒病毒成为目前唯一可以进行全面遗传操作的真菌病毒。利用低毒病毒作为探针来探测板栗疫病菌的致病力组成和毒力调节机制,己获得了一些很有意义的发现。本文介绍近几年低毒病毒及其与真菌相互作用的研究进展,包括低毒病毒的基因组和功能基因研究、低毒病毒和线粒体损害引起的板栗疫病菌低毒力机制、板栗疫病菌的RNA沉默系统以及低毒病毒抗RNA沉默的机制。低毒病毒／板栗疫病菌系统已经成为研究病毒与宿主相互作用以及病原真菌致病机理的很好的模式系统。  相似文献   

19.
Kinetics of chlorophyll degradation and color loss in heated broccoli juice   总被引：2，自引：0，他引：2  
Degradation of chlorophyll in broccoli juice occurred at temperatures exceeding 60 degrees C. Chemical analysis revealed that degradation of chlorophyll a and b to pheophytin a and b, respectively, followed first-order kinetics and that chlorophyll a was more heat sensitive than chlorophyll b. Temperature dependencies of chlorophyll a and b degradation rate constants could be described by Arrhenius equations with activation energies (E(a)) of 71.04 +/- 4.89 and 67.11 +/- 6.82 kJ/mol, respectively. Objective greenness measurements, using the -a value as the physical property, together with a fractional conversion kinetic analysis, indicated that green color degradation followed a two-step process. Kinetic parameters for the first degradation step were in accordance with the kinetic parameters for pheophytinization of the total chlorophyll content, as determined by chemical analysis (E(a) approximately 69 kJ/mol). The second degradation step, that is, the subsequent decomposition of pheophytins, was characterized by an activation energy of 105.49 +/- 4.74 kJ/mol.  相似文献   

20.
Glucovanillin was extracted from green pods and simultaneously transformed to vanillin by a combination of enzyme activities involving cell wall degradation and glucovanillin hydrolysis. The reaction is best carried out with 47.5% v/v aqueous ethanol solution during 8 h at 70 degrees C, in a two-step enzymatic reaction using Viscozyme followed by Celluclast, two commercial enzymatic products containing mainly pectinase and cellulase activities, respectively. The extractive reaction proceeded with high efficiency with an amount of extracted vanillin 3.13 times higher than the one obtained with the Soxhlet method. The classical curing/extraction process results in 1.1-1.8 g of vanillin/100 g of dry pods. It is concluded that the enzymatic reaction may substitute the microbial process involved in tissue fermentation previous to vanillin extraction with the simultaneous hydrolysis of glucovanillin.  相似文献   

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