Electrochemical Studies of Intramolecular Electron Transfer in Laccase from Trametes versicolor

This paper describes electrochemical behavior of laccase from the fungus Trametes versicolor. The issues related to discrimination of the redox potentials corresponding to copper centers T1 and T2/T3 in the active site and possible mechanism of intramolecular electron transfer have been discussed. The electron‐transfer rate constant for laccase immobilized on carbon electrode is 3.4 s^?1. The bioelectrocatalytic activity of the enzyme was studied in the presence of 1,4‐hydroquinone (HQ). The kinetics of HQ oxidation is very fast (K_M=3.8 μM). However, the catalytic activity of laccase in the presence of high concentration of HQ decreases drastically. It is suggested that the T2/T3 copper center is able to accept electrons from HQ molecules directly via intramolecular channel.     

从野生与突变株云芝IBL-04提纯锰过氧化物酶及其表征

由野生及突变株云芝IBL-04制得细胞外锰过氧化物酶(MnPs),并经过硫酸铵沉淀、透析、离子交换和凝胶渗透层析法等步骤提纯.纯化的酶在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)上于43 kDa区域呈现单一谱带,它适宜的pH值和温度分别为5.0和40°C.突变株MnPs表现出比野生株MnPs更宽的活性pH值范围和更高的热稳定性.从所选突变株所得纯化的MnPs表现出与野生株MnP几乎相同的电泳性质、稳态动力学、金属离子和EDCs降解效率.该生物酶与Mn2+一起催化的反应速率最快,但最高的亲和性对应于ABTS、甲氧基羟苯基乙二醇、4-氨基苯酚和活性染料. Mn2+和Cu2+可显著提高MnPs的活性,但Zn2+, Fe2+, EDTA和半胱氨酸则会不同程度地抑制其活性, Hg2+是最强的活性抑制剂.所有来源的MnPs均可有效催化EDCs、壬基苯酚和二氯苯氧氯酚降解,处理3 h可除去80%以上,在MnPs-介质体系中可进一步提高到90%.综上,云芝MnPs生物酶具有较高的pH适用性和热稳定性、独特的Michaelis-Menten动力学参数和高的EDCs去除效率等特点,因而有望工业化应用.     

Laccase from Trametes versicolor

The enzyme laccase was produced by the white-rot fungus Trametes versicolor in repeated batches cultures with immobilized mycelium. Two different culture conditions were used. Enzymes produced were evaluated regarding their stability a thigh temperatures (55°C and 65°C) and at alkaline conditions (pH 7.0 and pH 8.0) having in view the application of these enzymes in biobleaching of hardwood Kraft pulp. Biobleaching experiments were divided in two parts, enzymatic prebleaching followed by chemical bleaching. In the enzymatic prebleaching the enzyme laccase was used at two conditions of pH and temperature, whereas the reaction time was fixed at 1h in all pretreatments. In the chemical bleaching the DEDED and DEpDED sequences were used. The enzyme action was evaluated by Kappa number, viscosity, and brightness at the end of bleaching sequences. There were obtained values of Kappa numbers lower than control assays, viscosities compatible with industrial pulps, and brightness higher than controls, when pulps were pretreated for 1 h with laccase at pH 8.0 and 55°C.     

Constituents from the Fruiting Bodies of Trametes cubensis and Trametes suaveolens in Vietnam and Their Anti-Inflammatory Bioactivity

It is reported that various fungi have been used for medicine and edible foods. The tropical Trametes genus is popular and well-known in Vietnam for its health effects and bioactivities. In this study, the fruiting bodies of the edible fungi T. cubensis and T. suaveolens were collected in Vietnam. The preliminary bioactivity screening data indicated that the methanol extracts of the fruiting bodies of T. cubensis and T. suaveolens displayed significant inhibition of superoxide anion generation and elastase release in human neutrophils. Therefore, the isolation and characterization were performed on these two species by a combination of chromatographic methods and spectrometric analysis. In total, twenty-four compounds were identified, and among these (1–3) were characterized by 1D-, 2D-NMR, and HRMS analytical data. In addition, the anti-inflammatory potentials of some purified compounds were examined by the cellular model for the inhibition of superoxide anion generation and elastase release in human neutrophils. Among the isolated compounds, (5,14), and (19) displayed significant anti-inflammatory potential. As the results suggest, the extracts and isolated compounds from T. cubensis and T. suaveolens are potential candidates for the further development of new anti-inflammatory lead drugs or natural healthy foods.     

Transformation of textile dyes by white-rot fungus Trametes versicolor

We have investigated transformation of eight industrial dyes by a whiterot fungus, Trametes versicolor. The fungus was found to decolorize Reactive Golden Yellow R, Procion Red, Reactive Violet 5, Reactive Blue 28, and Ponceau Red 4R at an initial dye concentration of 80 ppm within 72 h of incubation, whereas it took 5 d to completely decolorize Reactive Black 5 (40 ppm). However, it did not significantly decolorize Reactive Red 152 and Novatic Blue BC S/D. During decolorization in liquid medium, laccase and manganese-independent peroxidase (MiP) activities were detected in culture filtrate of T. versicolor. Dye-decolorizing activity of the culture was found to be associated with H₂O₂-dependent activity of the culture filtrate. Furthermore, dye-decolorizing activity of the culture filtrate was not influenced by Mn²⁺ or veratryl alcohol, thus suggesting a role of extracellular MiP in decolorization of synthetic dyes by T. versicolor.     

Anti-inflammatory and free radial scavenging activities of the constituents isolated from Machilus zuihoensis

A new biflavonol glycoside, quercetin-3-O-β-D-glucopyranoside-(3'→O-3')-quercetin-3-O-β-D-galactopyranoside (9), together with eight known compounds was isolated for the first time from the leaves of Machilus zuihoensis Hayata (Lauraceae). The structure of compound 9 was elucidated by various types of spectroscopic data analysis. Analysis of the biological activity assay found that compound 9 showed significant superoxide anion scavenging activity (IC?? is 30.4 μM) and markedly suppressed LPS-induced high mobility group box 1 (HMGB-1) protein secretion in RAW264.7 cells. In addition, the HMGB-1 protein secretion was also inhibited by quercitrin (3), ethyl caffeate (6), and ethyl 3-O-caffeoylquinate (7) treatment. In the LPS-stimulated inducible nitric oxide synthase (iNOS) activation analysis, two known compounds, quercetin (1) and ethyl caffeate (6), were found to markedly suppress nitric oxide (NO) production (IC?? value, 27.6 and 42.9 μM, respectively) in RAW264.7 cells. Additionally, it was determined that ethyl caffeate (6) down-regulated mRNA expressions of iNOS, IL-1β, and IL-10 in the LPS-treatment of RAW264.7 cells via a suppressed NF-kB pathway. These results suggested for the first time that the new compound 9 and other constituents isolated from M. zuihoensis have potential anti-inflammatory and superoxide anion scavenging effects. These constituents may be useful for treating various inflammatory diseases.     

Anti-inflammatory and anticancer activities of erythrodiol-3-acetate and 2,4-di-tert-butylphenol isolated from Humboldtia unijuga

Abstract

Humboldtia unijuga Bedd., endemic to Agasthyamala in Western Ghats in India, is traditionally used by local Kani tribes for chicken pox, head ache and snake bite. This study reports the isolation of erythrodiol-3-acetate (HU-1) and 2,4-di-tert-butylphenol (HU-2) from H. unijuga roots and their anti-inflammatory and anticancer activities in macrophage, skin and breast cancer cell lines. Effects of HU-1 and HU-2 treatments (50, 100?µg/mL) on gene expression profiles of pro-inflammatory cytokines TNFα, IL-6 and IL-1β, and apoptosis genes p53 and caspase 7 were studied. HU-2 exerted a significantly superior anti-inflammatory effect compared to HU-1 in all three pro-inflammatory genes. HU-2 showed a superior dose dependent anticancer effect through activation of p53 gene over HU-1 in MCF-7 cells. HU-1 exhibited a dose dependent effect on caspase 7 gene in both cell lines while HU-2 was more effective in A431. HU-2 has potential for development as a novel anti-inflammatory and anticancer agent.     

Novel Chromone Derivatives from Marine Fungus Aspergillus versicolor Isolated from the Sponge Xestospongia exigua

In the continuation of our research for the bioactive secondary metabolites from the sponge-associated fungi, the bioassay guiding fractionation led to the isolation of six new secondary metabolites with unusual skeleton based on chromone ring system from the inoculated fungus Aspergillus versicolor, that had been isolated from fresh samples of marine sponge Xestospongia exigua, collected along coast line of Bali, Indonesia in 1997. The basic skeleton of those compounds possessed an oxohexacy…     

Pulp bleaching using laccase from trametes versicolor under high temperature and alkaline conditions

Kraft pulp was delignified using laccase produced by the white rot fungusTrametes versicolor immobilized in solid support under specific conditions. The stability tests showed that this enzyme was stable for 6 h at 55°C and pH 8.0, allowing its use under pH and temperature conditions very close to those used in industrial bleaching. In this work, unbleached hardwood Kraft pulp was submitted to prebleaching using 2 U laccase/g pulp basis. Reaction time, temperature, and pH of the enzymatic treatment were investigated. Good results regarding Kappa number reduction, selectivities, and high viscosities were obtained when prebleaching was performed for 1 h at temperature of 55©C and pH 8.0 followed by alkaline extraction and ECF bleaching sequences.

     

Biotransformation of Aspen Lignin by the Fungus Trametes villosus

Quantitative ¹ H and ¹³ C NMR spectroscopies demonstrate that biotransformation of aspen wood by the fungusTrametes villosusresults in oxidation and destruction of lignin with cleavage of C-C alkyl-alkyl bonds in side chains and partial demethoxylation in addition to cleavage of lignocarbohydrate bonds. New C _ar -O-C bonds form while lignin is being destroyed at alkyl-alkyl bonds. Cleavage of rings and destruction of C _ar -C bonds was not observed.     

A Steroidal Glycoside from Cynanchum versicolor Bunge

Cynanchum versicolor Bunge, one species of the well known Chinese herbal medicine “paiwei”, has been used as an antifebrile and diuretic agent for a long time in China1. Previous phytochemical studies in this species have reported the occurrence of some…     

New chemical constituents from the fruits of Zanthoxylum armatum and its in vitro anti-inflammatory profile

Chemical investigations on the fruits of Zanthoxylum armatum Roxb. (Rutaceae) led to the isolation of two new constituents characterised as 2α-methyl-2β-ethylene-3β-isopropyl-cyclohexan-1β, 3α-diol (1) and phenol-O-β-D-arabinopyranosyl-4′-(3″, 7″, 11″, 15″-tetramethyl)-hexadecan-1″-oate (2) along with known compounds m-methoxy palmityloxy benzene (3), acetyl phenyl acetate (4), linoleiyl-O-α-D-xylopyranoside (5), m-hydroxyphenoxy benzene (6) and palmitic acid (7). The chemical structures were established with the help of physical, chemical and spectroscopic methods. The anti-inflammatory potential of isolated compounds 1 and 2 was evaluated using in vitro target-based anti-inflammatory activity in LPS-stimulated primary peritoneal macrophages isolated from mice. Production of pro-inflammatory cytokines (TNF-α and IL-6) was significantly inhibited by the treatment of isolated compounds 1 and 2 in a dose-dependent manner.     

Four Novel Hydropyranoindeno-Derivatives from Marine Fungus Aspergillus versicolor

In the previous report on bioactive secondary metabolites from the sponge-associated fungus Aspergillus versicolor (Vuill) Tirab, the bioassay guiding fractionation led to isolation of six new compounds with unusual skeleton based on chromone ring system from the inculated fungus which was isolated from fresh samples of marine sponge Xestospongia exigua, collected along coast line of Bali, Indonesia in 19971. In the continuation of our chemical investigation on the marine fungus, four compou…     

A new pregnane steroid from cultures of Aspergillus versicolor

A new pregnane steroid, named aspergillon A (1), together with two known compounds, (22E,24R)-ergosta-5,7,22-trien-3β-ol (2) and (22E, 24R)-ergosta-4,6,8(14),22-tetraen-3-one (3) were isolated from cultures of the tin mine tailings-associated fungus Aspergillus versicolor. The new structure and absolute configuration were determined with the help of extensive spectroscopic analyses and quantum chemical calculations of the electronic circular dichroism (ECD) spectra.     

Composite magnetic particles as carriers for laccase from Trametes versicolor

In this paper we report a study of laccase immobilisation on different kinds of carrier particles. The immobilisation of enzyme on the particle surface with respect to the immobilisation efficiency and the properties of the immobilised enzymes is discussed. The immobilisation of laccase on polystyrene particles bearing reactive beta-diketone groups is characterised by high efficiency, but grafting of the enzyme increases the stability of the colloidal system, which makes the separation/purification procedure difficult. Additionally, the extreme colloidal stability of the immobilisates hinders the application of such particles with immobilised enzymes in some applications where the recycling of the enzyme should be performed. It has been found that hybrid PS-AAEM particles equipped with maghemite show similar immobilisation efficiency to that of their analogues without maghemite and can additionally be manipulated in magnetic fields. The activity of the immobilised laccase is much higher in the pH region 5-7 and the temperature range 50-70 degrees C as compared with that of the free enzyme. Immobilised enzymes also exhibit much better storage stability.     

New polyhydroxylated metabolite derived from biotransformation of diosgenin by the white-rot fungus Coriolus versicolor

<正>Microbial transformation of diosgenin(1) was carried out with the white-rot fungus,Coriolus versicolor.A new polyhydroxyl metabolite,(25R)-spirost-5-ene-3β,7β,21-triol(2),was obtained as a result of hydroxylation.Its structure was elucidated on the basis of 1D and 2D NMR as well as HR-ESI-MS spectroscopic analysis.     

Antimicrobial activity and molecular docking studies of a novel anthraquinone from a marine-derived fungus Aspergillus versicolor

A novel anthraquinone, 2-(dimethoxymethyl)-1-hydroxyanthracene-9,10-dione (1), together with nine known compounds (2–10), were isolated from the fermentation of Aspergillus versicolor derived from deep sea sediment. Their structures were established through spectroscopic methods. Compound 1 exhibited strong inhibitory activities against MRSA ATCC 43300 and MRSA CGMCC 1.12409 (with MIC values of 3.9 and 7.8 μg/mL respectively) and moderate activities against tested strains of Vibrio (with MIC values ranging from 15.6 to 62.5 μg/mL). Compound 1 was subjected to molecular docking studies for inhibition of topoisomerase IV and AmpC β-lactamase enzymes indicating its usefulness as antimicrobial agent.     

Versicorin,a new lovastatin analogue from the fungus Aspergillus versicolor SC0156

A new lovastatin analogue versicorin (1), together with three related compounds, decumbenones A (2) and B (3) and versiol (4), were isolated from mycelial solid cultures of Aspergillus versicolor SC0156. Their structures were elucidated on the basis of MS data and NMR spectroscopic analysis. The new compound versicorin (1) possesses a hexahydro-2H-naphtho[1,8-bc]furan moiety, which is a rare type of the lovastatin-analogous compounds. A hypothetical biosynthetic pathway for compounds 1–4 was proposed.