Lignin peroxidase is involved in the biobleaching of manganese-less oxygen-delignified hardwood kraft pulp by white-rot fungi in the solid-fermentation system

Biobleaching of manganese-less oxygen-delignified hardwood kraft pulp (E-OKP) by the white-rot fungi Phanerochaete sordida YK-624 and P. chrysosporium was examined in the solid-state fermentation system. P. sordida YK-624 possessed a higher brightening activity than P. chrysosporium, increasing pulp brightness by 13.4 points after seven days of treatment. In these fermentation systems, lignin peroxidase (LiP) activity was detected as the principle ligninolytic enzyme, and manganese peroxidase and laccase activities were scarcely detected over the course of treatment of E-OKP by either fungus. Moreover, a linear relationship between brightness increase and cumulative LiP activity was observed under all tested culture conditions with P. sordida YK-624 and P. chrysosporium. These results indicated that LiP is involved in the brightening of E-OKP by both white-rot fungi.     

糙皮侧耳和鞘脂菌NS7对多环芳烃污染土壤的生物强化与协同作用

【背景】真菌和细菌被认为在多环芳烃污染土壤生物修复过程中发挥协同作用,目前在真实土壤体系中开展真菌-细菌协同降解研究较少。【目的】研究真菌和细菌对不同种类多环芳烃降解的差异及对蒽和苯并[a]蒽的生物强化与协同作用。【方法】选用多环芳烃降解真菌和细菌各一株,在液体纯培养体系下分析它们对不同种类多环芳烃降解的差异,在土壤体系中采用放射性同位素示踪技术研究2种微生物对蒽和苯并[a]蒽的生物强化与协同作用。【结果】供试细菌鞘脂菌NS7能够很好地降解低环种类多环芳烃,以蒽作为唯一碳源时可以将其完全降解,在复合污染条件下对菲、蒽、荧蒽、芘等降解效果突出(>90%),对苯并[a]芘降解效果较差(9.76%)。相比而言,供试真菌糙皮侧耳菌对苯并[a]芘具有更好的降解效果(21.18%),对低环多环芳烃降解效果明显不如降解菌NS7。在自然土壤中,蒽和苯并[a]蒽具有明显不同的矿化效率,分别为18.61%和4.28%,在蒽污染土壤中加入鞘脂菌NS7并未显著提高蒽的矿化率(P>0.05),相比而言,苯并[a]蒽污染土壤中加入糙皮侧耳显著提高了污染物矿化效率(2.24倍),表明真菌和细菌在土壤环境...     

Fractionation of subcellular membranes of the secretory pathway from the peroxidase-producing white-rot fungus Phanerochaete chrysosporium

Abstract Mycelia from the basidiomycete Phanerochaete chrysosporium , producing lignin and manganese peroxidases, were homogenized and fractionated on a sucrose gradient. The main subcellular fungal membrane fractions were successfully separated. Lipid composition analyses of the isolated membranes as well as associated marker enzymes distribution gave evidence to similarities with membranes originating from plants. Lignin and manganese peroxidases were investigated by immunodetection in subcellular fractions. Our results show that lignin and manganese peroxidases are mainly associated with Golgi apparatus vesicles and, to a lesser extent, with endoplasmic reticulum and light density vesicles, but not with plasma membranes.     

Isolation and characterization of a white rot fungus <Emphasis Type="Italic">Bjerkandera</Emphasis> sp. strain capable of oxidizing phenanthrene

Strain BOL13 was selected from 18 fungal strains isolated from an oil-spill contaminated site in Oruro, Bolivia. It was identified as a basidiomycete with high homology to Bjerkandera. The fungus degraded 100 mg phenanthrene l⁻¹ at 0.17 mg l⁻¹ d⁻¹ at 30 °C at pH 7. During phenanthrene degradation, a maximum manganese peroxidase activity of 100–120 U l⁻¹ was measured after 10 days of incubation. The ability of Bjerkandera sp. to produce lignin-modifying enzymes and to oxidize phenanthrene under various pH and temperature conditions was confirmed.     

Simultaneous biodegradation ofp-cresol and phenol by the basidiomycetePhanerochaete chrysosporium

Summary The fungusPhanerochaete chrysoporium BKM-F-1767 was able to degrade high concentrations ofp-cresol (up to 150 mg L^–1) provided that glucose was added as a carbon and energy source and conditions favourable to ligninolytic enzyme activities were used, i.e. a nitrogen-limited medium. The fungus also simultaneously degradedp-cresol (50 mg L^–1) and phenol (50 mg L^–1) in a mixture at similar rates. Kinetics ofp-cresol biodegradation were almost identical whether the compound was tested individually or in a mixture with phenol.     

Lignin-modifying enzymes from selected white-rot fungi: production and role from in lignin degradation

Abstract: White-rot fungi produce extracellular lignin-modifying enzymes, the best characterized of which are laccase (EC 1.10.3.2), lignin peroxidases (EC 1.11.1.7) and manganese peroxidases (EC 1.11.1.7). Lignin biodegradation studies have been carried out mostly using the white-rot fungus Phanerochaete chrysosporium which produces multiple isoenzymes of lignin peroxidase and manganese peroxidase but does not produce laccase. Many other white-rot fungi produce laccase in addition to lignin and manganese peroxidases and in varying combinations. Based on the enzyme production patterns of an array of white-rot fungi, three categories of fungi are suggested: (i) lignin-manganese peroxidase group (e.g. P. chrysosporium and Phlebia radiata ), (ii) manganese peroxidase-laccase group (e.g. Dichomitus squalens and Rigidoporus lignosus ), and (iii) lignin peroxidase-laccase group (e.g. Phlebia ochraceofulva and Junghuhnia separabilima ). The most efficient lignin degraders, estimated by ¹⁴CO₂ evolution from ¹⁴C-[Ring]-labelled synthetic lignin (DHP), belong to the first group, whereas many of the most selective lignin-degrading fungi belong to the second, although only moderate to good [¹⁴C]DHP mineralization is obtained using fungi from this group. The lignin peroxidase-laccase fungi only poorly degrade [¹⁴C]DHP.     

Effect of growth substrate,method of fermentation,and nitrogen source on lignocellulose-degrading enzymes production by white-rot basidiomycetes

The exploration of seven physiologically different white rot fungi potential to produce cellulase, xylanase, laccase, and manganese peroxidase (MnP) showed that the enzyme yield and their ratio in enzyme preparations significantly depends on the fungus species, lignocellulosic growth substrate, and cultivation method. The fruit residues were appropriate growth substrates for the production of hydrolytic enzymes and laccase. The highest endoglucanase (111 U ml⁻¹) and xylanase (135 U ml⁻¹) activities were revealed in submerged fermentation (SF) of banana peels by Pycnoporus coccineus. In the same cultivation conditions Cerrena maxima accumulated the highest level of laccase activity (7,620 U l⁻¹). The lignified materials (wheat straw and tree leaves) appeared to be appropriate for the MnP secretion by majority basidiomycetes. With few exceptions, SF favored to hydrolases and laccase production by fungi tested whereas SSF was appropriate for the MnP accumulation. Thus, the Coriolopsis polyzona hydrolases activity increased more than threefold, while laccase yield increased 15-fold when tree leaves were undergone to SF instead SSF. The supplementation of nitrogen to the control medium seemed to have a negative effect on all enzyme production in SSF of wheat straw and tree leaves by Pleurotus ostreatus. In SF peptone and ammonium containing salts significantly increased C. polyzona and Trametes versicolor hydrolases and laccase yields. However, in most cases the supplementation of media with additional nitrogen lowered the fungi specific enzyme activities. Especially strong repression of T. versicolor MnP production was revealed.     

Biodegradation of polycyclic aromatic hydrocarbons in soil around Mathura oil refinery,India

Six polycyclic aromatic hydrocarbons [naphthalene, anthracene, phenanthrene, pyrene, chrysene and benzo(a)-pyrene] were detected in soil receiving effluents from an oil refinery. Biodegradation studies revealed a time-dependent disappearance of these polycyclic aromatic hydrocarbons when they were added to soil samples: naphthalene disappeared completely in 60 days, whereas phenanthrene, anthracene, pyrene, chrysene and benzo(a)pyrene decreased by 87%, 34%, 21%, 5% and 40%, respectively, in 120 days.B.T. Ashok and J. Musarrat were and S. Saxena is with the Interdisciplinary Biotechnology Unit, A.M.U., Aligarh-202002, Uttar Pradesh, India. K.P. Singh is with the Environmental Chemistry Section, Industrial Toxicology Research Centre, M.G. Road, Lucknow-226001, Uttar Pradesh, India. B.T. Ashok is now with the Department of Biochemistry, J.N. Medical College, A.M.U., Aligarh-202002, Uttar Pradesh, India. J. Musarrat is now with the Department of Radiology and Blochemistry Program. The Ohio State University, Columbus, OH 43210, USA.     

Phenanthrene biodegradation and the interaction of <Emphasis Type="Italic">Pseudomonas putida</Emphasis> BS3701 and <Emphasis Type="Italic">Burkholderia</Emphasis> sp. BS3702 in plant rhizosphere

The interaction between the strains degrading polycyclic aromatic hydrocarbons, Pseudomonas putida BS3701 and Burkholderia sp. BS3702, was studied in the course of phenanthrene degradation in plant rhizosphere. Strain BS3702 was shown to accumulate 1-hydroxy-2-naphthoic acid (which is toxic for plants); it was then utilized by strain BS3701, which thereby increased the resistance of plants to the pollutant and to the toxic intermediate. With this type of interaction (cooperation), the efficiency of phenanthrene degradation was noted to decrease.     

Methanogenic bacteria as a key factor involved in changes of town gas stored in an underground reservoir

Growth of Phanerochaete chrysosporium in a nitrogen-limited medium buffered with sodium acetate, instead of the commonly used 2,2-dimethylsuccinate (DMS), resulted in quantitative and qualitative differences in the production of various extracellular lignin peroxidases (LIPs) and manganese-dependent peroxidases (MNPs) involved in lignin degradation. The results indicate that production of LIPs and MNPs can be selectively enhanced by manipulation of culture conditions. Partial N-terminal analyses of the major LIPs and MNPs have made it possible to assign a specific protein to the specific genes and cDNAs that have been reported recently. The LIPs and MNPs differed widely in their ability to decolorize various dyes that are known to be degraded by the lignin degrading enzyme system of P. chrysosporium.     

Potential of Trametes trogii culture fluids and its purified laccase for the decolorization of different types of recalcitrant dyes without the addition of redox mediators

Trametes trogii BAFC 463 culture fluids (containing 110 U ml⁻¹ laccase; 0.94 U ml⁻¹ manganese peroxidase), as well as its purified laccase were capable of decolorizing azoic, indigoid, triphenylmethane, anthraquinonic and heterocyclic dyes, in the absence of redox mediators. Six dyes: RBBR, Indigo Carmine, Xylidine, Malachite Green, Gentian Violet and Bromophenol Blue were almost completely degraded (more than 85% decolorization after 1 d) by either laccase or T. trogii itself in culture, proving the role of the enzyme in dye decolorization. The purified laccase also decolorized 65% of Fast Blue RR and 30% of Azure B and Methylene Blue after 24 h. The use of redox mediators significantly increased the decolorization rates (90% decolorization of Azure B after 1 h). 1-hydroxybenzotriazole resulted the best redox mediator, but the natural mediator p-hydroxybenzoic acid also demonstrated its efficiency for dye decolorization. Due to their ability to decolorize recalcitrant dyes without addition of redox mediators, high laccase activities, high thermostability and efficient decolorization at 70 °C and pH 7.0, even in the presence of high concentrations of heavy metals (100 mM Cu⁺², Pb⁺² or Cd⁺²) or in a synthetic dyebath, T. trogii culture fluids could be effectively used to decolorize synthetic dyes from effluents.     

Enzymatic degradation of mould stains on paper analysed by colorimetry and DRIFT-IR spectroscopy

Melanin is chemically and by physical characteristics very similar to lignin, a major constituent of wood, and therefore ligninolytic enzymes of white-rot fungi were tested for their ability to selectively degrade melanin. Melanin degradation was studied both in liquid suspensions of melanin and on melaninised paper samples. Liquid suspension samples were tested for changes in their chemical composition (appearance and relative representation of functional groups and chemical bonds) with FTIR spectrometry. Changes in colour of melaninised paper samples were investigated with a colorimeter. Effectiveness of the treatment (bleaching) was determined as a change in lightness (ΔL). Melanin was oxidised in the liquid suspensions, and the intensity of modification varied depending on the procedure employed. The most pronounced changes in melanin were observed in laccase-1-hydroxybenzotriazole (HBT) treatment at heightened air pressure. The most prominent discoloration of the melaninised paper samples (and no visually detectable damage to the integrity of the paper) was, like in the case of the liquid suspensions, observed after laccase-HBT treatment.     

Novel heme-containing enzyme possibly involved in lignin degradation by the white-rot fungus Junghuhnia separabilima

Abstract The white-rot fungus Junghuhnia separabilima (Pouz.)Ryv, showed high levels of laccase production in cultures supplemented with veratric acid. Laccase, lignin peroxidase and an unknown peroxidase were separated from the extracellular culture fluid using anion-exchange FPLC. Three laccase species, three lignin peroxidases and a novel heme-containing protein were characterized by gel electrophoresis and isoelectric focusing. The new hemoprotein has a molecular mass of 44 kDa, isoelectric point of 3,4 and pH optimum of 5.5 for oxidation of o -dianisidine in the presence of H₂O₂. However it oxidised diaminobenzidine and guaiacol in the absence of H₂O₂. Veratryl alcohol and phenol red were not substratesfor this enzyme with or without addition of H₂O₂ and Mn(II). In addition the enzyme did not produce H₂O₂.     

微生物降解多环芳烃的研究进展

多环芳烃(PAHs)是具有严重危害的环境污染物质。介绍PAHs的降解菌,降解机理和PAHs的生物修复方面的研究进展。土壤中PAHs的生物修复被认为是解决污染的有效方法,目前,菲的生物降解途径已经比较清楚,但对结构更为复杂的多环芳烃研究较少。文章还对消除环境中多环芳烃的相关生物技术提出展望。     

Remediation of benzo[a]pyrene and chrysene-contaminated soil with industrial hemp (Cannabis sativa)

The phytoremediation, with industrial hemp (Cannabis sativa), of a Hawaiian silty clay soil contaminated with two polycyclic aromatic hydrocarbons (PAHs), chrysene and benzo[a]pyrene, was studied. Hemp showed a very high tolerance to the contaminants. The growth rates of hemp, compared with control, in soils fortified with chrysene and benzo[a]pyrene at concentrations of each varying from 25 to 200 micrograms/g were consistently above 100%. The plants grew from seed for 45 days in soil fortified with PAHs at concentrations of 25, 50, and 75 micrograms/g. Controls were pots with contaminated soil but no plant. PAHs levels were significantly reduced in all pots (control and seeded pots), expect for one set at a high concentration of chrysene, which may be due to uneven spiking. A time course study over 28 days was done to monitor changes of microbial count and levels of chrysene. Little changes were observed for the total microbial count in the soil, and the concentration of chrysene in the soil decreased slightly in the pots containing plants. However, the chrysene levels in those pots were consistently lower than those in the pots without plants.     

污染土壤中多环芳烃的共代谢降解过程

1　前　言多环芳烃是一类普遍存在于环境中的重要有机污染物 ,因其致癌性、致畸性、致突变性而被认为是危险物质。由于其水溶性低 ,辛醇 水分配系数高 ,因此 ,该类化合物易于从水中分配到生物体内、沉积层中。土壤成为多环芳烃的重要载体 ,多环芳烃污染土壤的生物修复也因此倍受关注。多环芳烃在土壤中有较高的稳定性 ,其苯环数与其生物可降解性明显呈负相关关系。很少有能直接降解高环数多环芳烃的微生物。研究表明 ,高分子量的多环芳烃的生物降解一般均以共代谢方式开始[1 3] 。共代谢作用可以提高微生物降解多环芳烃的效率 ,改变微生物碳…     

Biodegradation of polycyclic aromatic hydrocarbons by Pichia anomala

Pichia anomala 2.2540, isolated from soil contaminated by crude oil, degraded naphthalene, dibenzothiophene, phenanthrene and chrysene, both singly and in combination. The yeast degraded 4.5 mg naphthalene l(-1) within 24 h. Phenanthrene was degraded after a lag of 24 h. When a mixture of all four polycyclic aromatic hydrocarbons was treated at either 0.1-1.6 mg l(-1) or 3.1-5.3 mg l(-1), naphthalene was completely degraded first within 24 h, followed by phenanthrene and dibenzothiophene after 48 h. Chrysene, which remained in the mixture even after 96 h, could be degraded along with naphthalene. Chrysene at 0.7 and 1 mg l(-1), in the presence of 4.3 and 65 mg naphthalene l(-1), respectively, was removed within 96 h.