The role of Mn-dependent peroxidase in dye decolorization by static and agitated cultures ofIrpex lacteus

Dye decolorization capacity of two white-rot fungi, Irpex lacteus and Phanerochaete chrysosporium, was compared in N-limited liquid cultures. The agitated cultures showed lower ability to decolorize azo dyes Reactive Orange 16 and Naphthol Blue Black than static cultures. Similar effect was also observed with other structurally different synthetic dyes. The effect of surfactants on the decolorization process is discussed. A significant increase in the Reactive Orange 16 decolorization by the agitated I. lacteus cultures was observed after adding 0.1% Tween 80, following a higher Mn-dependent peroxidase production. The in vitro dye decolorization using the purified enzyme proved its decolorization ability.     

Evaluation of a new fungus <Emphasis Type="Italic">Ceriporia lacerate</Emphasis> strain P2—its ability to decolorize Alizarin Red and Methyl Orange

A new fungus Ceriporia lacerate P2 which belongs to family Polyporaceae was evaluated for its ability to decolorize two different dyes Alizarin Red and Methyl Orange. Different parameters such as incubation time, pH, carbon source, nitrogen source and carbon/nitrogen regimes were used to find out the optimum medium for Ceriporia lacerate P2 on its ability of decolorization. The results show that the fungus had different ability to decolorize the two tested dyes. For Alizarin Red, the most suitable medium was at pH 3 and the best carbon and nitrogen source were sucrose and ammonium nitrate. While for Methyl Orange, the optimum medium was at pH 7–9 and the best carbon and nitrogen source were sucrose and urea.     

Evaluation of synthetic dye decolorization capacity in Ischnoderma resinosum

The little studied white rot fungus Ischnoderma resinosum was tested for its ability to decolorize seven different synthetic dyes. The strain efficiently decolorized Orange G, Amaranth, Remazol Brilliant Blue R, Cu-phthalocyanin and Poly R-478 on agar plates and in liquid culture at a relatively high concentration of 2–4 and 0.5–1 g l⁻¹, respectively. Malachite Green and Crystal Violet were decolorized to a lower extent up to the concentration of 0.1 g l⁻¹. Decolorization capacity of I. resinosum was higher than that in Phanerochaete chrysosporium, Pleurotus ostreatus or Trametes versicolor. In contrast with these thoroughly examined fungi, I. resinosum was able to degrade a wide spectrum of chemically and structurally different synthetic dyes. I. resinosum also efficiently decolorized dye mixtures. In liquid culture, Orange G and Remazol Brilliant Blue R were decolorized most rapidly; the process was not affected by different nitrogen content in the media. Shaken cultivation strongly inhibited the decolorization of Orange G.     

White-rot fungus <Emphasis Type="Italic">Ganoderma</Emphasis> sp.En3 had a strong ability to decolorize and tolerate the anthraquinone,indigo and triphenylmethane dye with high concentrations

The ability of the white-rot fungus Ganoderma sp.En3 to decolorize different kinds of dyes widely applied in the textile and dyeing industry, including the anthraquinone dye Remazol Brilliant Blue R (RBBR), indigo dye indigo carmine and triphenylmethane dye methyl green, was evaluated in this study. Ganoderma sp.En3 had a strong capability of decolorizing high concentrations of RBBR, indigo carmine and methyl green. Obvious reduction of Chemical Oxygen Demand was observed after decolorization of different dyes. Ganoderma sp.En3 had a strong ability to tolerate RBBR, indigo carmine and methyl green with high concentrations. High concentrations of RBBR, indigo carmine and methyl green could also be efficiently decolorized by the crude enzyme of Ganoderma sp.En3. Different redox mediators such as syringaldehyde, acetosyringone and acetovanillone could enhance the decolorization capability for higher concentration of indigo carmine and methyl green. Different metal ions had little effect on the ability of the crude enzyme to decolorize indigo carmine and methyl green. Our study suggested that Ganoderma sp.En3 had a strong capability for decolorizing and tolerating high concentrations of different types of dyes such as RBBR, indigo carmine and methyl green.     

Uptake of reactive textile dyes by Aspergillus foetidus

An isolated fungus, Aspergillus foetidus was found to effectively decolorize media containing azo reactive dyes namely, Drimarene dyes. The extent of color removal was greater than 95% within 48 h of growth of the fungus. The entire color was found to be strongly bioadsorbed to the rapidly settling fungal biomass pellets without undergoing significant biotransformation. Our investigations reveal that the process of decolorization is concomitant with the exponential growth phase of the fungus and has requirement for a biodegradable substrate such as glucose. The fungus was also able to decolorize media containing mixture of dyes to an extent of 85% within 72 h of growth. Kinetic analyses of fungal decolorization indicate that the process is time dependent and follows first order kinetics with respect to initial concentration of dye. The rates of color uptake (k values) decrease to a significant extent with increasing initial concentrations of dye. The fungus was able to grow and decolorize media in the presence of 5 ppm of chromium and 1% sodium chloride. An alternate and cheaper carbon source such as starch supported the growth and decolorization process. These results suggest that dye uptake process mediated by A. foetidus has a potential for large-scale treatment of textile mill discharges.     

The implication of Dichomitus squalens laccase isoenzymes in dye decolorization by immobilized fungal cultures

The study focuses on the production of ligninolytic enzymes and dye degradation capacity of Dichomitus squalens immobilized on polyurethane foam (PUF) or pine wood (PW) in a fixed bed reactor at a laboratory scale (working volume of 0.6l). Immobilization of fungal cultures on pine wood improved eminently laccase production in comparison to the liquid cultures. Immobilized D. squalens was able to decolorize an anthraquinone dye Remazol Brilliant Blue R and an azo dye Reactive Orange 16, however, only a limited decolorization of Copper(II)phthalocyanine dye was observed in both types of reactor cultures. The involvement of a laccase activity in dye decolorization was suggested. Further, two different chromatographical forms of laccases, Lc1 and Lc2, were isolated from PW cultures of D. squalens using a fast, two step FPLC method. Enzymes revealed identical molecular masses of 68 kDa (estimated by SDS-PAGE) and similar pI's, however, they differed in their catalytic properties such as pH dependence of the activity and ABTS oxidation rates. In this study, we demonstrated different dye decolorization capacities of Lc1 and Lc2 as well.     

Decolorization of simulated textile dye baths by crude laccases from Trametes hirsuta and Cerrena unicolor

In this study crude laccases from the white‐rot fungi Cerrena unicolor and Trametes hirsuta were tested for their ability to decolorize simulated textile dye baths. The dyes used were Remazol Brilliant Blue R (RBBR) (100 mg/L), Congo Red (12.5 mg/L), Lanaset Grey (75 mg/L) and Poly R‐478 (50 mg/L). The effect of redox mediators on dye decolorization by laccases was also assessed. C. unicolor laccase was able to decolorize all the dyes tested. It was especially effective towards Congo Red and RBBR with 91 and 80% of color removal in 19.5 h despite the fact that simulated textile dye baths were used. Also Poly R‐478 and Lanaset Grey were partially decolorized (69 and 48%, respectively). C. unicolor laccase did not need any mediators for removing the dyes. However, T. hirsuta laccase was only able to decolorize simulated Congo Red and RBBR dye baths (91 and 45%, respectively) in 19.5 h without mediators. When using mediators the decolorization capability was enhanced substantially, e.g. Poly R‐478 was decolorized by 78% in 25.5 h. On the whole, both laccases showed potential to be used in industrial applications.     

Decolorization of Victoria blue by the white rot fungus, <Emphasis Type="Italic">Phanerochaete chrysosporium</Emphasis>

Synthetic textile dyes are among the most dangerous chemical pollutants released in industrial wastewater streams. Recognizing the importance of reducing the environmental impact of these dyes, the ability of the white rot fungus Phanerochaete chrysosporium to decolorize various textile dyes was investigated. This fungus decolorized 6 of the 14 structurally diverse dyes with varying efficiency (between 14% and 52%). There was no discernable pattern of decolorization even among dyes of the same chemical class, suggesting that attack on the dyes is relatively non-specific. Among the three dyes which showed >40% decolorization, Victoria Blue B (VB) was chosen for further analysis because the ability of the fungus to decolorize VB was nearly independent over a relatively broad concentration range. Blocking lignin peroxidase (LiP) and manganese peroxidase (MnP) production by the fungus did not substantially affect VB decolorization. Inhibition of laccase production by adding various inhibitors to shaken cultures reduced VB decolorization significantly suggesting a role for laccase in VB decolorization. When sodium azide and aminotriazole were used to inhibit endogenous catalase and cytochrome P-450 oxygenase activities, there was 100% and 70% reduction in VB decolorization, respectively. Adding benzoate to trap hydrogen peroxide-derived hydroxyl radicals resulted in 50% decolorization of VB. Boiling the extracellular fluid (ECF) for 30 min resulted in approximately 50% reduction in VB decolorization. Collectively, these data suggest that laccase, and/or oxygenase/oxidase and a heat-stable non-enzymatic factor, but not Lip and MnP, play a role in VB decolorization by P. chrysosporium.     

多孔菌Trichaptum abietinum 1302BG自然条件下对合成染料刚果红和酸性品红的高效降解

选用杭州竹林土壤分离并筛选能够降解多种类型染料的真菌。经大量筛选发现一株编号为1302BG的真菌能够在固体培养基上分解所测试的全部9种染料(苯胺蓝、刚果红、橙黄G、甲基红、甲基橙、结晶紫、酸性品红、番红花红、碱性品红、甲基紫)。经形态学和分子生物学方法鉴定, 该菌1302BG为冷杉附毛孔菌(Trichaptum abietinum)。在液体培养基中研究了pH、温度、碳源、氮源、碳氮源组合、碳氮源浓度等参数对该菌脱色效果的影响, 以寻找最适最经济的脱色条件。在液体培养基中研究表明, 冷杉附毛孔菌1302BG既能在酸性又能在碱性条件下有效分解2种测试染料(酸性品红和刚果红)。该真菌能以仅含有0.5 g/L淀粉和0.05 g/L硫酸铵的经济、环境友好的培养基为底物, 能在灭菌和非灭菌(自然)的条件下高效脱色, 在24 h内对2种染料的脱色率均在90%以上。紫外/可见光谱及微核试验分析显示, 该菌脱色主要是以生物降解为主, 2种染料经该菌分解后的毒性也同时大大降低。这些优异特点显示了该菌具有非常广阔的工业染料废水处理应用潜力。     

Capacity of Irpex lacteus and Pleurotus ostreatus for decolorization of chemically different dyes

The rate and efficiency of decolorization of poly R-478- or Remazol Brilliant Blue R (RBBR)-containing agar plates (200 μg g⁻¹) were tested to evaluate the dye degradation activity in a total of 103 wood-rotting fungal strains. Best strains were able to completely decolorize plates within 10 days at 28 °C. Irpex lacteus and Pleurotus ostreatus were selected and used for degradation of six different groups of dyes (azo, diazo, anthraquinone-based, heterocyclic, triphenylmethane, phthalocyanine) on agar plates. Both fungi efficiently degraded dyes from all groups. Removal of RBBR, Bromophenol blue, Cu-phthalocyanine, Methyl red and Congo red was studied with I. lacteus also in liquid medium. Within 14 days, the following color reductions were attained: RBBR 93%, Bromophenol blue 100%, Cu-phthalocyanine 98%, Methyl red 56%, Congo red 58%. The ability of I. lacteus to degrade RBBR spiked into sterile soil was checked, the removal being 77% of the dye added within 6 weeks. The capacity of selected white rot fungal species to remove efficiently diverse synthetic dyes from water and soil environments is documented.     

裂褶菌菌株G18对孔雀石绿染料的脱色优化

通过对兔眼蓝莓幼果组织中分离得到的内生真菌G18进行形态特征、ITS序列和系统进化分析鉴定菌株G18为裂褶菌Schizophyllum commune。同时,对菌株G18产生的3种木质素降解酶进行监测,发现G18菌株可以分泌漆酶、木质素过氧化物酶和锰过氧化物酶。为明确裂褶菌G18对染料的脱色能力,利用裂褶菌G18对固体条件下8种染料进行脱色能力的检测,筛选出较易脱色的染料后,对该染料的脱色条件进行优化。结果表明,裂褶菌G18对8种染料均可以脱色,对孔雀石绿染料的脱色效果最好。裂褶菌G18对孔雀石绿的脱色优化结果为pH 7.0、20.0g/L淀粉、1.0g/L尿素、1.0g/L硫酸锌、接菌量9片（d=5.0mm）。     

藜芦醇和吐温80对白腐菌产木质素降解酶的影响及在偶氮染料脱色中的作用

担子菌PM2在限氮液体培养下,分泌木质素过氧化物酶和锰过氧化物酶;藜芦醇、吐温 80的补充,提高了该菌锰过氧化物酶的产生,获得的最大锰过氧化物酶Mnp酶活为254.2u/L、190.2 u/L,分别是对照的3.4倍和2.5倍。选择三种偶氮染料,在染料体系下,进一步分析藜芦醇、吐温 80对担子菌PM2产过氧化物酶及染料脱色的影响。结果表明,担子菌PM2分泌的锰过氧化物酶Mnp与染料脱色有关,脱色程度受其分子结构特征影响;吐温80的补充,更有利于染料的脱色降解,48h后三种染料均可达到80%以上的脱色率。     

烟管孔菌G11对活性染料的脱色

本研究从未成熟的有机蓝莓表皮分离、纯化得到一株白腐真菌G11,通过对菌株G11的形态特征、ITS序列同源性比对以及系统发育分析,鉴定菌株G11为一株烟管孔菌Bjerkandera adusta。菌株G11可以产生木质素过氧化物酶、漆酶和锰过氧化物酶等木质素降解酶。菌株G11对8种不同染料的脱色效果显示其对活性染料的脱色效果最好,脱色率达到90%所需时间最短。以菌株G11为研究对象,研究其对不同浓度的活性黑和活性红的脱色能力,结果表明：菌株G11对活性红和活性黑具有显著的脱色能力。在脱色15d时,菌株G11对浓度为10、50、100、250、500mg/L活性红的脱色率分别为99%、98%、95%、94%和92%;对浓度为10、50、100、250、500mg/L活性黑的脱色率分别为98%、97%、95%、93%和90%。     

Implication of mycelium-associated laccase from Irpex lacteus in the decolorization of synthetic dyes

The white rot fungus Irpex lacteus is able to decolorize such synthetic dyes as Reactive Orange 16 and Remazol Brilliant Blue R. Here, we demonstrate that this type of dye decolorization is mainly related to a laccase-like enzyme activity associated with fungal mycelium. In its bound form, the enzyme detected showed a pH optimum of 3.0 for the oxidation of ABTS, DMP and guaiacol, and a pH of 7.0 for syringaldazine. The highest enzymatic activity was obtained with ABTS as substrate. Enzyme activity was fully inhibited with 50mM NaN(3). Depending on the chemical structure of dyes, redox mediators had a positive effect on the dye decolorization by fungal mycelium. Enzyme isolated from fungal mycelium was able to decolorize synthetic dyes in vitro.     

Decolorization of synthetic dyes by<Emphasis Type="Italic">Pleurotus ostreatus</Emphasis> isolates differing in ligninolytic properties

The ability to decolorize four synthetic dyes (Phenol Red, Evans Blue, Eosin Yellowish and Poly B411) in fivePleurotus ostreatus strains (a parental strain and four isolates derived from it) was determined. Two of the isolates had markedly higher and other two substantially lower production of ligninolytic enzymes and hydrogen peroxide that the parental strain. Like the parental strain, the higher-producing isolates were able to decolorize all the tested dyes, but not to a higher extent than the parental strain. In contrast, two lower-producing isolates exhibited slow decolorization, which was incomplete even at the end of cultivation. Evans Blue and Eosin Yellowish strongly suppressed the growth of the strains, while Phenol Red and Poly B411 induced none or only a very slight growth reduction.     

Biodegradation and detoxification of dyes and industrial effluents by Ganoderma weberianum B-18 immobilized in a lab-scale packed-bed bioreactor

White-rot fungi are considered to be promising biotechnological tools to complement or replace the current technologies for the treatment of effluents from textile production plants. The aim of this work was to investigate the decolorization capacity of Ganoderma weberianum B-18 in solid state fermentation with sugarcane bagasse as a substrate and ligninolytic inducer as well as to decolorize and detoxify industrial effluents by this strain in a laboratory scale packed-bed bio-reactor. The results demonstrated that G. weberianum B-18 indeed showed to possess decolorization capacity in solid state fermentation with sugarcane bagasse supplemented with synthetic dyes. Moreover, fungal biomass of G. weberianum B-18 immobilized in sugarcane bagasse in a packed-bed bioreactor was shown to efficiently decolorize and detoxify different dyes and authentic industrial effluents in semi-continuous conditions. In this decolorization process, laccase enzymes secreted by the fungus played the main role. Hence, a packed-bed reactor with G. weberianum B-18 immobilized in sugarcane bagasse seems to be a suitable system for the further development of an efficient bioprocess for large-scale treatment of dye-containing wastewaters.     

Decolorization of an anthraquinone-type dye using a laccase formulation

Decolorization of the dye Remazol Brilliant Blue R (RBBR) was studied, as it is representative of an important class of recalcitrant anthraquinone-type dyes. For this purpose a commercial laccase formulation (CLF) containing laccase, a redox mediator and a non-ionic surfactant was used. Small molecular weight components were removed from the CLF by gel filtration, which made it possible to compare the effect of its laccase alone. Apart from slightly better thermostability of the CLF as compared with the laccase alone, the pH and temperature profiles were similar regardless of the presence of the small molecular weight components. The laccase alone did not decolorize RBBR. A small molecular weight redox mediator (HBT) was necessary for decolorization to occur. A comparison of the kinetics of RBBR decolorization using the CLF and its laccase alone is reported. Provided that a redox mediator is included, it is suggested that laccase may be suitable for the wastewater treatment of similar anthraquinone dyes.     

黄孢原毛平革菌对RBBR脱色降解及其降解机制

为研究白腐真菌对蒽醌染料的生物降解机制,以白腐真菌黄孢原毛平革菌为脱色降解菌株,分析了蒽醌染料活性艳蓝KN-R(RBBR)的浓度、金属离子及脱色参数对染料脱色的影响;采用紫外-可见光谱、红外光谱、气相色谱-质谱(GC-MS)分析和植物种子毒性实验进行降解产物分析,以揭示RBBR可能的降解路径及其产物的毒性结果表明:在p...     

Efficient decolorization and detoxification of the sulfonated azo dye Reactive Orange 16 and simulated textile wastewater containing Reactive Orange 16 by the white-rot fungus Ganoderma sp. En3 isolated from the forest of Tzu-chin Mountain in China

The sulfonated azo dye Reactive Orange 16 is the commonly used representative of reactive dyes, but is hard to be degraded by some conventional treatment methods. In order to develop more efficient and more cost-effective treatment methods for degrading this recalcitrant dye, the capability of the white-rot fungus Ganoderma sp. En3 isolated by our laboratory to decolorize and detoxify Reactive Orange 16 was investigated in this study. Ganoderma sp. En3 had a strong ability to decolorize high concentrations of Reactive Orange 16 and simulated textile wastewater containing Reactive Orange 16 in submerged cultures. Decolorization of Reactive Orange 16 and its simulated dye effluents by this fungus resulted in the decrease of phytotoxicity. Ganoderma sp. En3 had strong adaptability and tolerance to high concentrations of Reactive Orange 16. Compared with some previous research, Ganoderma sp. En3 was superior to some other fungal strains reported previously in the rate and extent of decolorizing Reactive Orange 16. It was also found that the real textile wastewater could be efficiently decolorized by Ganoderma sp. En3 in submerged cultures. The crude enzyme produced by Ganoderma sp. En3 could also efficiently decolorize Reactive Orange 16 and simulated textile wastewater under in vitro conditions.     

Stock dynamics of Cleisthenes herzensteini in the central and southern Yellow Sea

Anthropogenic activities and environmental changes have had a significant effect on the fishery ecosystem, biological characteristics, and population dynamics of marine fishes. Overfishing threatens the sustainability of many populations. We evaluated changes in the biological characteristics, distribution, and abundance of Cleisthenes herzensteini using bottom trawl survey data collected from 1985 to 2010 in the central and southern Yellow Sea. The dominant body length of C. herzensteini during spring was 80–160 mm in 1986, 60–160 mm in 1998, and 41–80 mm and 111–170 mm in 2010. During summer, the dominant body length was 80–180 mm and 130–169 mm in 2000 and 2007, respectively. During autumn, the dominant body length was 60–160 mm, 100–180 mm, and 90–149 mm in 1985, 2000, and 2009, respectively. During winter, the dominant body length was 80–200 mm, 120–220 mm, and 100–200 mm in 1985, 1999, and 2010, respectively. The dominant body length decreased gradually from 1985 to 2010 (excluding spring, 2010), illustrating the “miniaturization” of the C. herzensteini population. Growth was significantly different between male and female individuals, with male individuals forming a “smaller-size type”. The sex ratio of C. herzensteini was relatively stable during spring and summer, but significantly different during autumn and winter. The diet of C. herzensteini also changed significantly from 1985 to 2010. During 1985–1986, the diet consisted primarily of Crangon affinis, Eualus sinensis and Gammaridae species. C. affinis, Engraulis japonicus, and Ammodytes personatus were dominant during 1998–2000, whereas C. affinis was the dominant prey species during 2009–2010. Thus, there was a clear decrease in dietary diversity, with a shift to benthos shrimp, particularly C. affinis, which accounted for 82.58% of the total diet (by weight) in 2010. The gastric vacuous rate also decreased in every season and the gonad developmental stage changed with each season. The distribution of C. herzensteini shifted northward and offshore and became more concentrated. The average catch per haul of C. herzensteini decreased in spring and autumn. The average catch per haul ranged from 1.44 kg h^-1 to 0.14 kg h^-1 in spring and the percentage by weight ranged from 6.53% to 1.28%. The average catch per haul ranged from 3.03 kg h^-1 to 0.26 kg h^-1 in autumn and the percentage by weight ranged from 8.00% to 0.60%. The average catch per haul increased significantly during summer, ranging from 0.18 kg h^-1 to 0.58 kg h^-1, with a percentage by weight of 0.03–0.80%. The average catch per haul was relatively stable in winter (around 1.00 kg h^-1), but the percentage by weight gradually increased during 1985–2010. Taken together, our results suggested that the population structure, diet composition, and distribution of C. herzensteini had been altered during the last three decades. To address this, it is essential to initiate measures to conserve the C. herzensteini resource.