白腐菌Trametes versicolor产漆酶发酵条件的优化

对白腐菌Trametesversicolor产漆酶的发酵条件进行了研究,结果表明摇瓶培养产漆酶的最佳培养基组成为:可溶性淀粉2g/L,氯化铵1.2g/L,KH2PO42g/L,MgSO4·7H2O0.5g/L,CaCl2·2H2O0.1g/L,VB10.001g/L,Tween802g/L,微量元素混合液7mL/L,愈创木酚0.015mmol/L,CuSO4·5H2O60μmol/L,pH3.5;最佳发酵条件为:在250mL三角瓶装50mL培养基,接种量(Φ8mm)2块,25℃,150r/min振荡培养10d时,漆酶活力达到862U/L,约是优化前的4倍。

白腐菌Trametes hirsuta SYBC-L19液态发酵水葫芦产漆酶

利用单因素法和响应面法对白腐菌Trametes hirsuta SYBC-L19利用水葫芦液态发酵产漆酶的培养基和培养条件进行了优化,确定了最佳培养基:水葫芦41.0 g/L,玉米浆粉50.8 g/L,葡萄糖30.0 g/L,CuSO4.5H2O 0.6 g/L,吐温-80 0.5 g/L,香兰素0.5 mmol/L,无机盐混合液50 mL/L。最佳培养条件为:培养温度30℃、初始pH值4.0、装液量80 mL/250 mL、接种体积分数8%,200r/min摇瓶培养8 d,酶活性达7 784 U/L,是优化之前的17.3倍。该研究为水葫芦等生物质资源的有效利用提供了参考。

菌株Trametes hirsuta zlh237发酵液对污染土壤中多环芳烃的降解及群落结构分析

【目的】利用中国科学院微生物研究所真菌学国家重点实验室分离获得的菌株Trametes hirsuta zlh237,研究生物强化(接种菌株T.hirsuta zlh237发酵液)对污染土壤中3种多环芳烃(PAHs)[Phenanthrene、Pyrene和Benzo[a]pyrene(BaP)]的降解效果,为该菌株在PAHs污染土壤中的应用提供科学依据。【方法】采用高效液相色谱(HPLC)检测7个处理土壤(接种菌株T.hirsuta zlh237发酵液的Phenanthrene、Pyrene和BaP污染土壤分别标记为PheBA、PyrBA和BaPBA,接种灭菌发酵液的污染土壤分别标记为Phe、Pyr和BaP,原始土壤标记为CK)中3种PAHs含量,利用ABTS法检测土壤中漆酶活性,并运用高通量测序技术分析修复后土壤中细菌群落结构的变化。【结果】菌株T.hirsuta zlh237在3种PAHs污染土壤中均能生长,接种菌株发酵液15 d后,3种PAHs均有一定降解,其中BaP降解效果最佳,降解率达33.99%;且菌株T.hirsuta zlh237在3种污染土壤中均能分泌漆酶。高通量测序Alpha多样性指数分析结果表明,污染土壤接种菌株T.hirsuta zlh237发酵液后,Chao1指数和Shannon指数显著增加(P<0.05),不同处理土壤样品的Chao1指数和Shannon指数排序均为:PheBA>PyrBA>BaPBA>CK>Phe>Pyr>BaP。Beta多样性的主成分分析(PCA)结果表明,接种菌株T.hirsuta zlh237发酵液能改变污染土壤细菌群落结构组成;在门分类水平上,污染土壤样品中变形菌门(Proteobacteria)为优势门;在属分类水平上,接种菌株T.hirsuta zlh237发酵液的污染土壤样品中鞘氨醇单胞菌属(Sphingomonas)为优势菌属,接种灭菌发酵液的污染土壤Phe和Pyr样品中苯基杆菌属(Phenylobacterium)为优势菌属,而BaP样品中假单胞菌属(Pseudomonas)为优势菌属。【结论】菌株T.hirsuta zlh237发酵液在PAHs污染土壤中能分泌漆酶,对3种PAHs均有一定的降解效果,且能改变污染土壤细菌群落结构组成,在一定程度上对PAHs污染土壤有较好的修复效果。

白腐菌Trametes sp. Lg-9粗酶液预处理对杨木SCMP性能的影响

研究了白腐菌Trametes sp. Lg-9粗酶液预处理对磺化化学机械浆(SCMP)性能的影响。结果表明:Lg-9粗酶液预处理可改善SCMP的打浆性能,降低打浆能量消耗,预处理后打浆的纤维平均长度增大,细小组分含量降低,宽度稍有增大。Trametes sp. Lg-9粗酶液预处理可提高SCMP白度,改善强度。Trametes sp. Lg-9粗酶液预处理的漂后浆纤维平均长度较长,细小组分含量较少,粗酶液用量大于8IU/g浆时,随着粗酶液用量的增大,纤维平均长度有所降低,细小组分含量有所提高。

Trametes trogii WT-1降解欧美杨107杨木质素的初步研究

通过对受毛栓菌侵染的欧美杨107杨木块木质素含量的测定,结合扫描电镜(SEM)观察和红外光谱(IR)分析方法,对WT-1降解欧美杨107杨木块的过程进行初步研究。结果表明:受WT-1的侵染,杨木块木质素含量降低,其木质素降解过程呈现出慢-快-慢的规律;SEM观察结果表明,WT-1对杨木块木质素的降解由胞腔向胞间层进行,其中射线和薄壁组织最先降解,木纤维及导管次之;IR分析表明,受WT-1的侵染杨木块木质素结构发生了变化,木质素芳香核之间通过碳-碳键(—C—C—)连接的空间网状结构被破坏,木质素苯环间的羰基、CH2结构、紫丁香基和愈疮木基等侧链部分被降解,苯环骨架变化明显。

白腐菌Trametes hirsuta BYBF产漆酶发酵条件的优化

本实验对白腐菌Trametes hirsuta BYBF产漆酶的发酵条件进行了研究。结果表明,Trametes hirsuta BYBF的最佳培养条件为:培养基初始pH为6.0,培养温度为28℃,碳源为蔗糖,氮源为硝酸铵和酒石酸铵的组合,三角瓶装液量为200/500mL,振荡培养14天左右,酶活达到最大值,为1.37IU/mL。

Trametes versicolor预处理棉秆RMP制浆性能的研究

对白腐菌Trametes versicolor预处理棉秆RMP制浆性能进行了研究。结果表明,Trametes ver-sicolor有强的产漆酶能力,能对棉秆有效降解。随Trametes versicolor预处理时间的增大,RMP成浆的得率和白度逐渐降低,强度性能有明显的提高;但过长的预处理时间会降低强度性能。预处理初期,随处理时间增大,纤维平均长度逐渐增大,细小组分含量逐渐降低;随着处理时间的进一步延长,纤维平均长度有所降低。Trametes versicolor处理时间的延长可改善磨浆性能。Trametes versicolor处理不同时间的漂白性能不同,处理时间为6d时,各光学性能指标变化很小,而强度有较大提高;当处理时间达到18d以后,白度明显低于对照样。

Purification and characterization of manganese peroxidases from native and mutant Trametes versicolor IBL-04

Extracellular manganese peroxidases （MnPs） produced by native and mutant strains of Trametes versicolor IBL‐04 （EB‐60, EMS‐90） were purified by ammonium sulphate precipitation and dialysis, followed by ion‐exchange and gel‐permeation chromatography. The purified enzymes elucidated a single band in the 43‐kDa region on sodium dodecyl sulphate‐polyacrylamide gel electrophoresis. The optimum pH and temperature of the purified enzymes were found to be 5.0 and 40 °C, respec‐tively. Mutant strain MnPs exhibited a broader active pH range and higher thermal stability than native MnP. Purified MnPs from selected mutants showed almost identical properties to native MnP in electrophoresis, steady‐state kinetics, and metal ion and endocrine‐disrupting compound （EDC） degradation efficiency. Although the fastest reaction rates occurred with Mn2＋, MnPs displayed the highest affinity for ABTS, methoxyhydroquinone, 4‐aminophenol and reactive dyes. MnP activity was significantly enhanced by Mn2＋and Cu2＋, and inhibited in the presence of Zn2＋, Fe2＋, ethylene‐diaminetetraacetic acid and cysteine to various extents, with Hg2＋ as the most potent inhibitory agent. MnPs from all sources efficiently catalyzed the degradation of the EDCs, nonylphenol and triclosan, removing over 80%after 3 h of treatment, which was further increased up to 90%in the presence of MnP‐mediator system. The properties of T. versicolor MnPs, such as high pH and ther‐mal stability, as well as unique Michaelis‐Menten kinetic parameters and high EDC elimination effi‐ciency, render them promising candidates for industrial exploitation.

The protective effect and crucial biological pathways analysis of Trametes lactinea mycelium polysaccharides on acute alcoholic liver injury in mice based on transcriptomics and metabonomics

Trametes lactinea mycelia polysaccharides(TLMPS)have a wide range of bioactivities.The potential mechanisms of action of TLMPS against acute alcohol-induced liver injury in vivo were investigated by analyzing the physical and chemical properties of TLMPS and its protective effects on a mouse model of alcoholic liver injury.TLMPS protected the liver against alcohol-induced injury,as evidenced by the reduced alcohol-induced elevation of the liver index,serum biochemical indices,and maintenance of hepatic morphology.Potential mechanisms were analyzed using transcriptome and metabolome analyses.The transcriptome data revealed the involvement of many differentially expressed genes in chemical carcinogenesis,drug metabolism,and metabolism of xenobiotics by cytochrome P450.The metabolome analysis revealed that TLMPS significantly regulated specific metabolites in the liver,including organic acids,lipids,nucleosides,and organic oxygen compounds.KEGG enrichment analysis revealed the signifi cant involvement of different metabolites in choline metabolism,ATP-binding cassette transporters,and glycerophospholipid metabolism.Assessment of the changes in gene expression and metabolites revealed the significantly different expression of several genes encoding key enzymes and metabolites in choline metabolism pathway.The collective fi ndings confi rmed that choline metabolism plays an important role in the protective effects of TLMPS against acute alcoholic liver injury.

Properties of crude laccase from Trametes versicolor produced by solid-substrate fermentation

A low-cost process for the production of laccases is necessary for a sustainable enzymatic wastewater treatment. Therefore, it is necessary to establish an easy and low-cost procedure for the production of laccase. In the present study the properties of crude laccase from Trametes versicolor produced by solid-substrate fermentation is investigated. The application of the enzyme for dye decolorization is also studied. Crude laccase from the studied culture established maximal activity at 45&#186;C. The enzyme retained over 90% of its activity in the temperature range 40- 47&#186;C and pH 4.5. The kinetic constants of the crude enzyme was also determined. In the presence of KCl, NaCl, CaCl2, MnSO4 and MgSO4, laccase demonstra- ted high stability—over 50% of its initial activity was still retained after 4-month incubation. Complete loss of enzymatic activity was observed in the presence of CuCl2, FeCl2, FeCl3 and NaN3 after 30 min of incubation. 100% decolorization by investigated crude laccase was completed in the case of Indigo Carmine for 4 h, Remazol Brilliant Blue R—for 6 h, Orange II— for 48 h and Congo Red—for 13 d.

Effect of laccase from Trametes versicolor on the oxidative stability of edible vegetable oils

The increasing demand of natural food from consumers has limited the use of traditional methods to control the oxidation of lipids,such as synthetic antioxidants and hydrogenation.Besides,it has been reported that the use of enzymes is efficient to eliminate dissolved oxygen in foods such as vegetable oils.Laccase is a polyphenol oxidase and the reduction of oxygen to water is accompanied by the oxidation,typically,of a phenolic substrate.Laccase have become important,industrially relevant enzymes that can be used for a number of diverse applications such waste detoxification,textile dye transformation,food technologic uses,biosensor and analytical applications,bioethanol production,among others.The target of this study was to evaluate the effect of laccase enzyme from Trametes versicolor,on the oxidative stability of sesame,chia,peanut and sunflower oils,measured through the peroxide value(PV)and conjugated dienes(K232)and trienes(K270).The samples of oil with laccase showed higher PV,K232 and K270 than their corresponding controls,under the conditions evaluated(room temperature and 60◦C).The results suggest that fungal laccase has an unexpected pro-oxidant effect on vegetable oils,possibly promoted by products derived from the oxidation of phenols by enzymatic action.

Bidirectional solid fermentation using Trametes robiniophila Murr. For enhancing efficacy and reducing toxicity of rhubarb (Rheum palmatum L.)

Objective:To investigate the potential of bidirectional solid fermentation of rhubarb (Rheum palmatum L.) for reducing its toxicity and enhancing its medicinal efficacy.Methods:The fungus Trametes robiniophila Murr.was inoculated into rhubarb.The chemical ingredients as well as antioxidant,antibacterial,and anticancer activities of fermented and unfermented rhubarb extracts were then determined.Results:After fermentation,levels of anthraquinone glycosides (purgative ingredients) decreased significantly,while the level of anthraquinone aglycone increased.The level of gallic acid was also reduced after fermentation.Ethanol extract of rhubarb (0.8 mg/mL) exhibited DPPH-scavenging activity of 7.6% ± 0.8% while the blank control (0.8 mg/mL rhubarb)showed 31.3% ± 2.0% activity.Antibacterial activities in fermented samples were found to be enhanced compared with unfermented samples and anticancer activity was evident at concentrations of 2000 and 5000 μg/mL.Conclusion:Bidirectional solid fermentation appears to be an effective processing method that can be used to improve the efficacy and reduce the toxicity of rhubarb.

Trametes sp.MA-X01漆酶对不同染料的脱色作用

目的:研究真菌Trametes sp.MA-X01所产漆酶对不同种类染料的脱色作用,考察反应体系中不同条件对脱色效率的影响;方法:以3种类型(偶氮类、杂环类、三苯甲烷类)6种染料为研究对象,分别是偶氮类染料伊文思蓝、酸性铬蓝K、杂环类染料酸性红94、碱性红2,三苯甲烷类染料亮绿SF、孔雀石绿。采用单因素实验方法,考察反应体系中添加不同介体、反应温度、给酶量和染料质量浓度对脱色率的影响;结果:真菌Trametes sp.MA-X01所产漆酶对不同结构的染料均有较好的脱色作用。在相同的条件下,反应体系中加入介体ABTS可以提高脱色效率;反应温度40℃时的脱色效果最好。但该酶对不同染料的降解能力不同,反应24h,伊文思蓝的脱色率为80.31%,酸性铬兰K的脱色率为59.77%,酸性红94的脱色率为47.19%,碱性红2的脱色率为30.77%,孔雀石绿的脱色率为91.24%,亮绿SF的脱色率为89.09%;结论:真菌Trametes sp.MA-X01所产漆酶对不同结构染料均有较好的脱色效果,在纺织染料废水以及环境污染物降解等方面有着较好的开发和应用前景。

Purification and characterization of a novel immunomodulatory protein from the medicinal mushroom Trametes versicolor

Bioactive proteins represent an important group of functional agents in medicinal mushrooms.Trametes versicolor(L.) Lloyd is a mushroom frequently used in traditional Chinese medicine for its anti-tumor and immunomodulatory activities.A new immunomodulatory protein from T.versicolor,named TVC,was purified by ammonium sulfate precipitation,ion-exchange chromatography and gel filtration chromatography.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the purified protein revealed a single band with a molecular weight of 15.0 kD.Native polyacrylamide gel analysis revealed a band at 30 kD,indicating that TVC exists in solution as a homodimer.Isoelectric focusing showed that TVC was an acidic protein with an isoelectric point of 4.0.TVC was found to lack carbohydrate modifications(based on periodic acid/Schiff staining) and it does not agglutinate mouse red blood cells,suggesting that TVC is not a lectin-like protein.Biological activity assays dem-onstrated that TVC can enhance the proliferation of splenocytes,while it has no stimulatory effects on CD4+and CD8+T cells. TVC markedly increases the proliferation of human peripheral blood lymphocytes in a dose-dependent manner and enhances the production of both nitric oxide and tumor necrosis factor-alpha by lipopolysaccharide-induced murine macrophages.The results indicate that TVC is an immunostimulant that can boost immune response.Comparison of the N-terminal amino acid residues and mass spectrometry results with the protein database revealed no homologous proteins.

粗毛栓菌Trametes hirsuta Lg-9预处理玉米秸秆皮制机械浆初步探讨

探讨Trametes hirsuta Lg-9预处理玉米秸秆皮机械法制浆,研究Trametes hirsuta Lg-9接种物的菌龄和培养方式,接种后按有无外通空气两种培养方式对秸秆皮机械浆性能的影响。结果显示,按无外通空气方式处理时,13天菌龄的接种物处理样强度性能优于26天菌龄接种物处理得到的秸秆皮浆;固态静置培养接种物处理样强度性能优于液态震荡培养接种物的处理样。与对照样相比,处理样的强度、白度和光散射系数分别降低50%～70%,1%～4%和11%～16%,光吸收系数几乎都高于对照样。处理后的秸秆皮磨浆能耗降低70%～85%。且磨浆能耗与处理后秸秆皮得率间线性相关不显著。

Extractum trametes robiniophila murr protects murine podocyte via actin cytoskeleton stabilization

The Chinese medicine Extractum trametes robiniophila murr(ETRM)is the extract of a type of fungus.Recent studies have suggested that ETRM efficiently improves the effacement of podocyte foot processes in adriamycin(ADR)-induced nephrotic rats.In the present study,we aimed to assess whether ETRM modulated the actin rearrangements of podocytes and involved signaling molecules,includingα-actinin-4 and nephrin.Podocytes were treated with ADR(0.5μmol/L),ADR(0.5μmol/L)+dexamethasone(Dex)(1μmol/L),ADR(0.5μmol/L)+ETRM(10 mg/mL).The F-actin in the podocytes was stained by fluorescent phallotoxins and observed by confocal microscopy.The expression levels ofα-actinin-4 and nephrin were tested by real-time PCR and Western blotting analysis.The administration of ETRM could significantly prevent ADR-treated podocytes from actin rearrangement.Both ETRM and Dex could stabilize podocyte actin cytoskeletons.Moreover,α-actinin-4 might act as a potential target for ETRM functionality in podocyte actin rearrangements.However,pretreatment with ETRM could not inhibit the up-regulation of nephrin as a result of ADR treatment.ETRM could improve the cytoskeleton stability in ADR-induced actin rearrangement of podocytes via regulatingα-actinin-4 expression.

基于ITS序列的栓菌属部分种的分子分类初步研究(英文)

栓菌属 Trametes 的一些近缘种宏观和微观形态学非常相近,传统分类学方法难于对其进行准确分类定位。测定了 34 个分类单元的 ITS(包括 5.8SrDNA)序列,并对得到的 43 个分类单元的 ITS 序列进行系统发生分析,构建了聚类分析树状图。该树状图显示,栓菌属类群与其他属类群明显分开,Trametes versicolor 聚类到一个高支持率的独立分支。形态学上定名为 T. hirsuta 和 T. pubescens 物种聚类到同一高支持率的独立分支,试验分析表明这两个种应视为同一物种。

栓菌420漆酶C基因的克隆、高效表达及重组酶的染料脱色潜能

根据漆酶铜结合保守区氨基酸序列设计简并引物,从新型漆酶合成菌株栓菌420(Trametes sp.420)基因组DNA扩增得到一新的漆酶同工酶基因(lacC)片段,应用长距离反向PCR技术获得其两端侧翼序列。克隆得到的lacC序列长3640bp,包括长2263bp的开放读码框及5′和3′-非编码区。lacC cDNA序列长1560bp,编码一519aa的多肽。推导的LacC蛋白序列内部存在有10个潜在的N-糖基化位点和4个铜原子结合区。将不含自身信号序列的lacC cDNA以pPIC9载体为媒介克隆到表达载体pPIC9K上,转化毕赤酵母(Pichiapastoris)GS115细胞。重组菌在含有0.3mmol/LCuSO4和0.8%丙氨酸的BMM培养基中20℃培养9d,重组漆酶(rLacC)产量达到1.62×104U/L。用发酵粗酶液对终浓度50mg/L的染料进行脱色实验,结果表明,6U/L的rLacC对测试的三甲基类和偶氮类染料具有良好的脱色作用,小分子介体ABTS和HBT能够提高rLacC对染料的脱色效率和脱色速度。

Characterization of recuperating talent of white-rot fungi cells to dye-contaminated soil/water

This study was purposed to explore the decolorization of dyes by fungi on either a soil or in a liquid medium and to determine the application through batch shaking system. Two commercial dyes were decolorized and studied with four fungal strains in three media. Fungal growth is the best in malt extract/glucose medium for all organisms. Decolorization of reactive blue 220 and methyl red was investigated in soil medium by Trametes versicolor. These dyes were removed 91% and 80% for methyl red and reactive blue 220 respectively(dye concentration; 100 mg·L^(-1)) by both organisms. Enzymatic activities were monitored. Laccase(Lac) and manganese peroxidase(Mn P) were detected. MnP enzyme had important role for the dye decolorization. This study demonstrates that it is possible to decolorize some synthetic dyes, which would be highly advanced for dye containing wastewater and soil. These applications could be used for dye bioremediation.

Microbial population in rotten living body of Salix matsudana

The microbial population in rotten living body of Salix matsudana caused by Trametes suaveolens (L.) was researched. 11 bacteria species (1 species of Bacillus, 2 species of Clostridium and 8 species of non-brood-cell bacteria), 1 species of Actinomyces that belongs to Lavendulac, 8 species of fungi and 6 species of Trichoderma were isolated from rotten trunk. The hyphae of Trametes suaveolens mainly existed between rotten sections and discoioration sections. In over-rotten section and healthy section the fungi (Trametes suaveolens) were not isolated. The microbes that lived in the discoloration section were the most in kinds and number and they were the pioneer microbes of wood rotting. Only after they dwelled in wood and eIiminated its rot-resistance,could wood-rotting fungi invade wood and caused wood-rotting.