南极土壤嗜冷真菌Chrysosporium sp.C_(3438)活性代谢产物C_(3438)A的分离及结构鉴别

以对精原细胞法是否有活性作为筛选模型 ,对南极土壤嗜冷真菌 Chrysosporium sp. C3 4 3 8经低温发酵的活性产物进行了深入的化学研究 ,首次从南极土壤微生物代谢产物中分离得到

Toxicity and bioaccumulation of heavy metals in Phanerochaete chrysosporium

The responses of the growth and metabolism activity of Phanerochaete chrysosporium （P. chrysosporium） to cadmium （Cd）, lead （Pb） and their combined pollution stress, were investigated in plate and liquid culture conditions. The diameter of colony, biomass ofP. chrysosporium, ligninolytic enzyme activities and bioaccumulation quantity of heavy metals were detected. The results indicated that Cd was more toxic than Pb to P. chrysosporium and the toxicity of Cd and Pb to P. chrysosporium was further strengthened under Cd＋Pb combined pollution in different culture conditions. Heavy metals Cd and Pb had indirect influence on the production of ligninolytic enzymes by directly affecting the fungal growth and metabolic activity, and by another way in liquid culture. In addition, the results provided an evidence of the accumulation of Cd and Pb on the mycelia ofP. chrysosporium.

Influence of Phanerochaete chrysosporium on degradation and preg-robbing capacity of activated carbon

The effect of Phanerochaete chrysosporium on degradation and preg-robbing capacity of activated carbon,which was used as a substitute of carbonaceous matter in carbonaceous gold ores,was studied.After 14 d treatment with Phanerochaete chrysosporium,the degradation rate of activated carbon reached 27.59%.The XRD and FTIR analyses indicate that Phanerochaete chrysosporium can distort the micro-crystalline structure of activated carbon,increase the number of oxygen-containing groups and aliphatics and make the aromatic structures be oxidized and exfoliated.The gold-adsorption tests show that Phanerochaete chrysosporium can reduce the preg-robbing capacity of activated carbon by 12.88%.This indicates that Phanerochaete chrysosporium is an available microorganism,and it can be employed to reduce the preg-robbing capacity of carbonaceous matter and improve the gold leaching rate.The combined effect of passivation,alkalization and oxidation of biological enzymes-free radicals of Phanerochaete chrysosporium on carbonaceous matter was also discussed.

气液环境下白腐真菌Phanerochaete chrysosporium在载体表面的附着性能研究

以白腐真菌Phanerochaetechrysosporium为研究对象,考察了该菌种在液相和气相环境中在陶粒、颗粒炭、玻璃珠、竹子、聚丙稀多面空心球、沸石上的附着和生长情况.研究结果表明,在液相环境中,白腐真菌Phanerochaetechrysosporium对竹子、沸石、陶粒、聚丙稀多面空心球有较好的附着能力,但菌体的生长量和形态特征受载体的影响而产生明显差异.在有玻璃珠和颗粒炭存在的液相环境中,均出现一定数量的菌丝小球,但在载体表面无附着现象.将附有菌体的载体置于气相环境中,发现白腐真菌在载体表面能够形成气生菌丝层.同时,显微镜下观察发现,菌体在沸石、陶粒和多面空心球表面呈绒毛状,质地松散,在竹子表面的菌体有明显的交织结构.

黄孢原毛平革菌(Phanerochaete chrysosporium)对Pb^(2+)的生物吸附特性及吸附机理

研究了非活性黄孢原毛平革菌(Phanerochaete chrysosporium)菌丝球对Pb2+的吸附特性及吸附机理.结果表明,菌丝球对Pb2+的吸附是单分子层吸附,Langmuir模型比Freundlich模型更适合于描述Pb2+在菌丝球上的吸附平衡.菌丝球吸附Pb2+的过程可以分为初始的快速吸附和随后的缓慢吸收2个阶段,并可以用二级动力学模型来描述.在菌丝球吸附Pb2+的过程中虽然存在粒内扩散,但它不是吸附过程的限速步骤.Pb2+的生物吸附过程是放热过程,其吸附活化能为-35.78kJ/mol,Pb2+的吸附热与化学反应热相近,为-44.45 kJ/mol.该生物吸附至少存在2种机理:菌体表面一些含氧基团对Pb2+的络合作用;Pb2+与细胞表面H+的离子交换.

黄孢原毛平革菌(Phanerochaete chrysosporium)对3种酚酸物质的降解作用

研究黄孢原毛平革菌(P.chrysosporium)对香草酸、阿魏酸、对羟基苯甲酸的降解作用。结果表明,300μg/mL的上述3种酚酸物质与黄孢原毛平革菌共培养2d后,3种酚酸物质的降解率分别达到73.48%、99.09%和54.74%。黄孢原毛平革菌发酵上清液对3种酚酸物质的降解能力较弱,作用2d后,降解率分别为13.28%,4.47%和5.43%,而该菌菌丝体降解能力很强,作用2d后3种酚酸物质的降解率分别达到65%,98.37%和48.69%。研究表明,黄孢原毛平革菌对这3种酚酸的降解主要在菌体表面或体内进行。

Phanerochaete chrysosporium降解碳质金矿中元素碳高关联度变量的筛选

采用Plackett-Burman设计法对Phanerochaete chrysosporium降解碳质金矿中元素碳的高关联度变量进行筛选。通过前期的单因素试验确定影响真菌降解碳质物的8个变量,分别为愈创木酚浓度、糊精浓度、吐温-80浓度、草酸浓度、过氧化氢浓度、矿浆浓度、真菌浓度和作用时间。通过两水平的Plackett-Burman设计法确定影响真菌降解元素碳的高关联度变量,分别为愈创木酚浓度、草酸浓度和矿浆浓度。愈创木酚是一种诱导剂。低浓度时,愈创木酚通过提高酶活性和诱导一些酶产生,促进元素碳的降解。高浓度时,酶产生和酶活性受抑制,不利于元素碳的降解。草酸通过调节降解体系的p H值影响菌体生长及酶活性。矿浆浓度会影响元素碳和真菌的有效接触面积以及降解体系的剪切力和传质效率。

NaCl对黄孢原毛平革菌(Phanerochaete chrysosporium)的损伤效应

染料中含有大最NaCl是影响黄孢原毛平革菌脱色效率的重要因素。为研究NaCl对黄孢原毛平革菌处理功能的影响，分别采用孢子和菌丝球与不同浓度的NaCl混合培养10d，以观察孢子生长和菌丝球的损伤效应，并利用透射电子显微镜和AFLP法对其进行细胞结构分析与DNA扩增，通过分析不同浓度NaCl对其生长及微观结构的影响、NaCl浓度与DNA相似性关系以及构建UPGMA相似性树状图等方法，评价NaCl对P.chrysosporium结构与功能的损伤效应。结果显示，3％NaCl对黄孢原毛平革菌影响较小，细胞结构保持完整，异常细胞量为14．2％，DNA变异率小，与空白的相似度达90％以上，表明黄孢原毛平革菌在3％的浓度范围内结构功能基本不受影响；5％NaCl使DNA相似度下降为71．4％，下降幅度最为显著，并且细胞内含物松散和出现胞浆空泡化趋势，异常细胞占有量为71．1％，说明3％～5％的浓度范围最易对P．chrysosporium的结构与功能产生不良影响；NaCl浓度≥8％可对黄孢原毛平革菌产生严重损伤，细胞变形严重，空泡化，DNA相似度降至67％以下，异常细胞量约90％，表明此浓度范围可使黄孢原毛平革菌基本丧失了原有的结构与功能。

Effect of nitrogen concentration in culture mediums on growth and enzyme production of Phanerochaete chrysosporium

Effect of different nitrogen concentration in the mediums on growth and enzyme production of Phanerochaete chrysosporium was studied when glucose concentration was 10 g/L. The results showed that the medium contained 0.8 g/L ammonium tartrate is the best. It not only supply abundant nutrients for the growth of Phanerochaete chrysosporium, which make mycelia the best grow compared with the other medium, but also produce higher manganese-dependent peroxidase(Mnp) and laccase(Lac) activity. In addition, it is observed that the variation of mycelia surface is related to ligninolytic enzyme secreted by Phanerochaete chrysosporium. When the surface of mycelium pellets appeared burs, it predicts secondary metabolism begin. This experimentation demonstrated that when the ratio of carbon and nitrogen in nitrogen limited medium is equal to 100∶8, growth and enzyme production of Phanerochaete chrysosporium is the best, it could achieve the maximum Mnp and Lac activity.

Polycyclic aromatic hydrocarbon biodegradation and extracellular enzyme secretion in agitated and stationary cultures of Phanerochaete chrysosporium

The extracellular enzyme secretion and biodegradation of polycyclic aromatic hydrocarbons （PAHs） were studied in agitated and shallow stationary liquid cultures of Phanerochaete chrysosporium. Veratryl alcohol and Tween80 were added to cultures as lignin peroxidase （LIP） and manganese peroxidase （MnP） inducer, respectively. Shallow stationary cultures were suitable for the production of enzyme, whereas agitated cultures enhanced overall biodegradation by facilitating interphase mass transfer of PAHs into aqueous phases. The use of a LiP stimulator, veratryl alcohol, did not increase PAH degradation but significantly enhanced LiP activity. In contrast, Tween80 increased both MnP secretion and PAH degradation in shallow stationary cultures. On the other hand, high PAH degradation was observed in agitated cultures in the absence of detectable LiP and MnP activities. The results suggested that extracellular peroxidase activities are not directly related to the PAH degradation, and the increased solubility rather than enzyme activity may be more important in the promotion of PAH degradation.

Biodegradation of Gaseous Chlorobenzene by White-rot Fungus Phanerochaete chrysosporium

Objective To evaluate the effect of white rot fungus Phanerochaete chrysosporium on removal of gaseous chlorobenzene. Methods Fungal mycelium mixed with a liquid medium was placed into airtight bottles. A certain amount of chlorobenzene was injected into the headspace of the bottles under different conditions. At a certain interval, the concentrations in the headspace were analyzed to evaluate the degradation of chlorobenzene by P. chrysosporium. Results The degradation effects of P. chrysosporium on chlorobenzene under different conditions were investigated. The difference in the optimum temperature for the growth of the fungi and chlorobenzene degradation was observed. The data indicated that a lower temperature （28℃） would promote the degradation of chlorobenzene than the optimum temperature for the growth of the fungi （37℃）. A low nitrogen source concentration （30 mg N/L） had a better effect on degrading chlorobenzene than a high nitrogen source concentration （higher than 100 mg N/L）. A high initial concentration （over 1100 mg/m3） of chlorobenzene showed an inhibiting effect on degradation by P chrysosporium. A maximum removal efficiency of 95% was achieved at the initial concentration of 550 mg/m3. Conclusion P. chrysosporium has a rather good ability to remove gaseous chlorobeuzene. A low nitrogen source concentration and a low temperature promote the removal of chlorobenzene by P. chrysosporium. However, a high initial chlorobenzene concentration can inhibit chlorobenzene degradation.

黄孢原毛平革菌(Phanerochaete chrysosporium)厚垣孢子降解阿特拉津的探究

阿特拉津(atrazine)是一类普遍存在于环境中且难降解的污染物.本文探究了黄孢原毛平革菌(Phanerochaete chrysosporium)厚垣孢子对阿特拉津降解的最佳条件,包括温度、摇床转速、初始培养基pH及接种量.并在大田土壤盆栽实验中,研究P.chrysosporium厚垣孢子和土壤土著微生物对土壤中阿特拉津的降解情况.结果表明:P.chrysosporium厚垣孢子可以有效去除阿特拉津,在33℃、转速为180r·min^(-1)、pH值为7.0、接种量是4g·L^(-1)时,去除效果最好,去除率达90.77%.土壤盆栽实验结果表明:施用P.chrysosporium厚垣孢子28d后,非灭菌土壤中阿特拉津去除率为97.8%,其中P.chrysosporium的降解贡献最为突出,去除能力为59.3%.而土著土壤微生物的去除率仅为20.7%,表明P.chrysosporium厚垣孢子对AT降解效果明显.

在大肠杆菌中直接克隆白腐丝状真菌-黄孢平革菌(Phanerochaete chrysosporium)基因启动子

利用启动子探钍型载体ｐＳＵＰＶ４ 直接在大肠杆菌细胞中克隆到多个来自白腐丝状真菌－黄孢平革菌（Ｐｈａｎｅｒｏｃｈａｅｔｅ ｃｈｒｙｓｏｓｐｏｒｉｕｍ ）的基因启动子片段．这些基因启动子片段能在大肠杆菌细胞中启动重组卡那霉素抗性基因（ｒｋａｎｒ）的表达，不同片段赋予宿主细胞以不同的卡那霉素抗性水平，最高的可达１０００μｇ／ｍ Ｌ，最低的则为５０μｇ／ｍ Ｌ．琼脂糖凝胶电泳显示这些重组质粒ＤＮＡ 中均有不同大小的插入片段，其范围是０．５～６．０ｋｂ．选取一个插入３．９ｋｂ 的重组质粒ｐＰＣ３３ 所作的Ｓｏｕｔｈｅｒｎ 杂交结果表明，插入片段以单拷贝形式存在于Ｐ．ｃｈｒｙｓｏｓｐｏｒｉｕｍ 的基因组中．当此片段５′－端被除去１．８ｋｂ 后，余下的２．１ｋｂ 片段可使融合的Ｋａｎｒ 基因的表达效率提高６０％ ．

P.chrysosporium降解处理多卤芳香污染物及染料污染物的研究进展

讨论了Phnerochaetechrysosporium木质素降解酶系降解木质素及其模型物的主要特性，催化降解多卤芳香污染物如多卤酚、多卤联苯二喔英以及各种染料的机制和特点。这种处理可以达到理想的降解和矿物化效果，但培养条件和污染物的具体结构对降解有明显影响；与细菌处理相比不产生新的有毒物质，有重要现实意义。P.chrysosporium处理多卤芳香污染物及染料污染物，具不可多得的优势，是解决上述污染潜在重要途径之一。

Manganese peroxidase production from cassava residue by Phanerochaete chrysosporium in solid state fermentation and its decolorization of indigo carmine

Bioconversion of lignocellulosic wastes to higher value products through fungal fermentation has economic and ecological benefits. In this study, to develop an effective strategy for production of manganese peroxidase(Mn P)from cassava residue by Phanerochaete chrysosporium in solid state fermentation, the stimulators of Mn P production were screened and their concentrations were optimized by one-at-a-time experiment and Box–Behnken design. The maximum Mn P activity of 186.38 nkat·g-1dry mass of the sample was achieved after 6 days of fermentation with the supplement of 79.5 mmol·L-1·kg-1acetic acid, 3.21 ml·kg-1soybean oil, and 28.5 g·kg-1alkaline lignin, indicating that cassava residue is a promising substrate for Mn P production in solid state fermentation. Meanwhile, in vitro decolorization of indigo carmine by the crude Mn P was also carried out, attaining the ratio of 90.18% after 6 h of incubation. An oxidative mechanism of indigo carmine decolorization by Mn P was proposed based on the analysis of intermediate metabolites with ultra-high performance liquid chromatography and gas chromatography tandem mass spectrometry. Using the crude Mn P produced from cassava residue for indigo carmine decolorization gives an effective approach to treat dyeing effluents.

Experimental research on Phanerochaete chrysosporium as coal microbial flocculant

The flocculation effect of the Phanerochaete chrysosporium on the coal slurry were studied by orthogonal experimental method in this study.The results of research show that the P.chrysosporium has a good effect on flocculating coal slurry.The optimum combination of the experimental is the P.chrysosporium,which is cultured in 2 days,mixed with 2 mL coagulant and 2 mL broth.The flocculant is a broken liquid with pH value of 6.The hightest flocculation ratio is 93.5%.The result of FTIR shows that the biological extraction of P.chrysosporium contains a lot of acidic polysaccharides that has the effect on flocculation.Microbial flocculant molecules and particles of coal slurry could be flocculated by‘‘absorption bridge’’.

Bio-dissolution of pyrite by Phanerochaete chrysosporium

The dissolution of pyrite was studied with Phanerochaete chrysosporium(P.chrysosporium).This fungus resulted in the dissolution of 18%iron and 33%sulfur.The oxidization layer was formed on the pyrite surface,which probably consisted of iron oxide,iron oxy-hydroxide,iron sulfate,elemental sulfur and mycelia.The electrochemical characteristics of pyrite were studied in the systems without and with P.chrysosporium.P.chrysosporium could accelerate the dissolution of pyrite by decreasing pitting potential and polarization resistance plus improving polarization current,corrosion potential and corrosion current density.The dissolution of pyrite is the combined effect of enzymes,hydrogen peroxide,ferric iron and organic acids.Enzymes attack the chemical bonds by free radicals.Organic acids dissolve pyrite by acidolysis and complexolysis.Enzymes and hydrogen peroxide play an essential role in this process.

Variation of peroxidase isoenzyme and biofilm of Phanerochaete chrysosporium in continuous membrane bioreactor for Reactive Brilliant Red X3-B treatment

The influence of a Reactive Brilliant Red X-3B （RBR X-3B） dye on the peroxidase isoenzyme ofPhanerochaete chrysosporium was determined, and the biofilm structure in a white rot fungal continuous membrane bioreactor （MBR） was also investigated by scanning electron microscope （SEM）. The variation of peroxidase isoenzyme and the decolorization rate in the continuous MBR were evaluated. The results showed that the 100 mg/L RBR X-3B could stimulate the production of the peroxidase isoenzyme in the shaking-flask culture. In addition, two new peroxidase isoenzyme bands with relative mobility （Rf） value of 0.27 and 0.28 appeared, but the activity was lower than the blank control of 11 d. In the continuous MBR, the system worked stably during the first 60 d, the main peroxidase isoenzyme bands existed and three new bands with Rf value of 0.10, 0.27, and 0.28 appeared. Meanwhile, the biofilm grew well and the average decolorization rate could reach 90.6%. But the bands of peroxidase isoenzyme decreased rapidly at day 65, only two bands with Rf value 0.24 and 0.26 existed, and the decolorization rate decreased to 78.3%. Therefore, 5 bottles of P. chrysosporium mycelial pellet were added into the MBR, and then the activity of the peroxidase isoenzyme and the decolorization rate had a slight recovery. Finally, the decolorization rate finally decreased to 75.2%. These results contribute to a comprehensive understanding of the variation of peroxidase isoenzyme and biofilm in continuous MBR by white rot fungi.

Construction of hybrid cell with Phanerochaete chrysosporium to degrade terephthalic acid wastewater——(I) phenotype evidence

The fungi Phanerochaete chrysosporium (PC) and Saccharomyces cerevisiae Y99 and the native bacterium YZ1 were the three parental strains for construction of hybrid cells through protoplast fusion to degrade terephthalic acid (TPA) wastewater. The results showed that the native bacterium YZ1 protoplasm could integrate with that of PC to form the hybrid cell Fhh and the fungus Y99 protoplasm also could integrate with that of Fhh to form the hybrid cell Fhhh. The protoplasts of YZ1 and Y99 could change the morphology of PC spore and mycelium for two times. The hybrid cell Fhhh got the best growth and degradation abilities in the wastewater. It suggested that the hybrid strains obtained from the inter\|kingdom protoplast fusion of the three parental strains could create potential for the purification of TPA wastewater.

Biosorption of lead by Phanerochaete chrysosporium in the form of pellets

The growth of Phanerochaete chrysosporium (ATCC 24725) in pellets was influenced by culture time, medium pH, C/N, surfactant concentration, spore number in inoculum, and shaking rate. The removal of Pb 2+ from aqueous solution by this kind of mycelial pellets was studied. The results indicated that many factors affected biosorption. These factors included pH, Pb 2+ concentration, co ion, adsorption time, and chemical pretreatments of biomass. Under optimum biosorption conditions(pH 4 5, 27℃, 16h), the highest lead uptake of 108 mg/g, was observed with mycelial pellets of 1 5-1 7 mm in diameter which were treated with 0 1 mol/L NaOH solution before adsorption. Pretreatment of biomass with NaOH further increased its biosorption capacity.