菌根真菌对环境污染物的降解、转化能力概述

菌根是高等植物与真菌形成的共生体，菌根真菌与许多微生物一样，也能有效的降解，转化的环境污染物，且有其自身的特点和优势。该文综合叙述了国内外在菌根真菌外在苗条奶真菌降解和转化木质素，多氯联苯，除草剂，有毒金属等环境污染物方面的研究进展，显示出菌根真菌是一大类具有较大应用潜力的，不容忽视的微生物群。     

白腐真菌F2的生长及产木质素降解酶特性的研究

考察了从我国生物资源中筛分出的白腐真菌F2的生长特性,采用多因素试验法研究了F2菌产木质素降解酶的特性;用统计学方法对试验数据进行了误差分析,并分析了多种影响因素对F2菌产木质素降解酶的影响.实验结果表明:F2菌在PDA培养基平板上和Tien＆Kirk培养基中都生长良好且生长速率较高;在自由悬浮振荡培养(不通氧气)条件下,F2菌产LiP的最高酶活可达60 5U·L-1,MnP的最高酶活可达263 4U·L-1,其产酶能力超过了黄孢原毛平革菌在自由悬浮振荡培养(不另外通入氧气)条件下的产酶能力.本研究还发现向培养基中添加的Cu2+浓度超过1μmol·L-1时会抑制F2菌产LiP,以往的文献未报道过Cu2+的这种抑制作用.     

几种纯培养外生菌根真菌对矿物油的降解效果

用紫外分光光度法对4种外生菌根真菌降解矿物油的能力进行了研究.结果表明,这4种外生菌根真菌都具有降解矿物油的能力.但在不同的培养基中其生物量和降解矿物油的能力显著不同,以有机氮作为氮源及提供少量葡萄糖作为启动碳源将大大提高其生物量及矿物油降解率.结果为应用菌根技术修复矿物油污染土壤及优化降解条件提供了依据.     

高效脱色菌群的脱色、产酶及群落分析

富集筛选到2组能够在开放条件下对多种染料和造纸废水高效脱色的混合菌群,以该混合菌群构建的生物膜反应器在连续运行条件下,对COD为750～1175mg/L、色度为200～320的染料废水进行脱色、降解,脱色率为80%～90%、COD去除率为69%～90%.该混合菌群能够在不同染料培养基中、造纸废水或染料废水中产生木质素降解酶系统中的1种或2种酶.分析表明,2组菌群中真菌与细菌的比例为6.8:1～51.8:1,优势微生物种类为真菌,从中分离出16株真菌,其中8株对活性红M-3BE具有明显脱色效果.     

白腐真菌酶学与分子生物学研究进展

白腐真菌对木质素降解的特异性生物学机制显示出其在复杂有机污染物降解中具有潜在应用前景。对白腐真菌酶学、自由基化学及分子生物学的国内外研究情况进行了较全面的介绍,以供国内同行参考。     

铝胁迫下外源钙对外生菌根真菌抗氧化保护酶活性的影响

采用液体培养方法,研究了4个不同菌株(Bo 02、Bo 15、Pt 715和Sl 08)的抗铝性及其在铝胁迫下加入0、0.25、0.5、1.0 mmol.-1L-1Ca2+处理后抗氧化保护酶活性的变化,旨在了解钙在缓解外生菌根真菌铝毒方面的作用.结果表明,不同菌株抗铝性不同,Pt 715和Sl 08的抗铝性强于Bo 02和Bo 15.铝胁迫可显著提高外生菌根真菌Bo 02 CAT和SOD活性、Bo 15SOD活性、Sl 08 CAT和POD活性,说明外生菌根真菌中这几种酶活性的提高与铝毒胁迫密切相关.4个菌株中,Bo 02酶活性对外源钙最敏感,外源钙对Bo 02铝胁迫的缓解作用最好.较高浓度的钙(≥0.5 mmol.L-1)可缓解或消除Sl 08铝胁迫造成的抗氧化酶活性上升.     

土壤微生物群落对多环芳烃污染土壤生物修复过程的响应

采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)方法,研究了土壤微生物群落多样性对生物表面活性剂强化的植物-微生物联合修复多环芳烃(PAHs)污染土壤的响应.结果表明,细菌群落的Shannon-Weaver指数修复前为3.17,修复后为3.24～3.45,多样性整体呈上升趋势,其中以植物-菌根真菌-降解菌处理最高,但各处理间无显著差异(P>0.05).聚类分析结果显示,植物、植物-鼠李糖脂、植物-菌根真菌和植物-菌根真菌-鼠李糖脂这4个处理的群落相似度在90%以上,植物-降解菌处理与这4个处理群落结构最近,此外,植物-降解菌-鼠李糖脂、植物-降解菌-菌根真菌-鼠李糖脂群落相似度在80%以上.通过测序比对,DGGE图谱上优势及特征性条带分别为Bacillus、Pseudomonas、Acidobacteria、Sphingmonas、Rhodopseudomonas、Firmicutes和Methylocytaceae等,可能是与PAHs降解密切相关的种属.生物表面活性剂强化的植物-微生物联合修复污染土壤过程中,在提高PAHs生物有效性基础上,改变了土壤微生物群落结构和丰度,从而可以有效提高PAHs的降解率.     

丛枝菌根真菌-植物联合修复石油污染土壤研究进展

丛枝菌根(Arbuscular mycorrhiza,AM)是由丛枝菌根真菌与植物根系所建立的一种互惠共生体,在自然界分布广泛,具有降解复杂有机污染物的能力.石油污染土壤中,利用土著的或外接的丛枝菌根真菌可以明显改善植物根际环境,提高土壤生物活性,从而加速土壤中石油污染物的降解.现有研究已证明丛枝菌根真菌修复石油污染土...     

植物内生真菌强化还田秸杆降解的研究

通过形态学和分子生物学相结合的方法 ,将内生真菌B3初步定为半知菌中的拟茎点霉属 (Phomopsissp .) .通过盆栽试验和扫描电镜观察表明 ,内生真菌B3有很好的降解秸秆的能力 ,尤其是对木质素的降解 ,而且对后季作物生长没有危害 .进一步研究木质素酶的性质表明 ,菌株B3分泌的漆酶有很好的耐热性 ,粗酶液在pH为 6 0— 8 0时 ,酶活较为稳定 .     

白腐真菌培养条件对其分泌木质素降解酶的影响

应用摇瓶试验研究了不同白腐真菌培养条件对其产酶的影响.通过控制不同转速、投加载体等方式考察了白腐真菌的产酶情况.结果表明,在悬浮培养方式下,转速为160r/min时获得的锰过氧化物酶最高(481.6U/L),其酶活是静置、120r/min条件下培养的65倍和43倍;投加载体后,载体固定化培养获得的木质素过氧化物酶酶活是悬浮培养的71倍,载体投加量对其产酶影响很大,只有当载体处于非浸没状态时才有利于酶活产生,否则酶活极低.因此,选择载体固定化—非浸没—振荡培养方式培养白腐真菌可获得更高的产酶量和酶活.     

Effects of culture conditions on ligninolytic enzymes and protease production by Phanerochaete chrysosporium in air

The production of ligninolytic enzymes and protease by Phanerochaete chrysosporium was investigated under different culture conditions. Different amounts of medium were employed in free and immobilized culture, together with two kinds of medium with different C/N ratios. Little lignin peroxidase （LIP） （〈 2 U/L） was detected in free culture with nitrogen-limited medium （C/N ratio： 56/2.2, in mmol/L）, while manganese peroxidase （MnP） maximum activity was 231 and 240 U/L in 50 and 100 ml medium culture, respectively. Immobilized culture with 50 ml nitrogen-limited medium gave the highest MnP and LiP production with the maximum values of 410 and 721 U/L separately on the day 5; however, flasks containing 100 ml nitrogen-limited medium only produced less MnP with a peak value of 290 U/L. Comparatively, carbon-limited medium （C/N ratio： 28/44, in mmol/L） was adopted in culture but produced little MnP and LiE Medium type had the greatest impact on protease production. Large amount of protease was produced due to glucose limitation. Culture type and medium volume influence protease activity corporately by affecting oxygen supply. The results implied shallow immobilized culture was a possible way to gain high production of ligninolytic enzymes.     

黄孢原毛平革菌在多种氨氮浓度下木质素降解酶的产生

在氨氮浓度0．0308-0．924g／L，下对黄孢原毛平革菌(Phanerochaete chrysosporium)在空气环境中进行自由悬浮振荡培养，在氨氮浓度0．0308-0．308g／L下可检测到木质素过氧化物酶活性，最高酶活为42．8U/L；在氨氮浓度0．0308g／L和0．154g/L下可检测到漆酶活性，最高酶活为35．0U／L，在氨氮浓度不低于0．154g/L时，葡萄糖消耗速率大致相当，明显高于在氨氮浓度0．0308g／L下的葡萄糖消耗速率；氨氮在葡萄糖耗尽时达到最低值，然后开始增大；木质素过氧化物酶和漆酶的活性峰与葡萄糖和氨氮的最大消耗基本对应。实验结果对木质素降解酶发酵和白腐真菌在环境工程中的直接应用具有启发意义。     

Cooperation between ligninolytic enzymes produced by superior mixed flora

Since the ability to degrade lignin with one kind of white-rot fungi or bacteria was very limited, superior mixed flora‘s ability to degrade lignin was investigated by an orthogonal experiment in this paper. The results showed that superior mixed flora reinforced the ability to degrade lignin, the degradation rates of both sample 9 and 10 were beyond 80% on the day 9. The cooperation between lignin peroxidase(LiP), Mn-dependent peroxidase(MnP) and laccase (Lac) for lignin degradation was also studied. By examining the activities of three enzymes produced by superior mixed flora, it was found that Lac was a key enzyme in the process of biological degradation of lignin but Lip was not; the enzyme activity ratios of Lac/MnP and Lac/LiP were significantly correlative with the degradation rate of lignin at the 0.01 level; and the ratio of MnP/LiP was an important factor affecting the degradation rate of lignin.     

白腐真菌木质素降解酶的反应器发酵

为了获得更高的白腐真菌木质素降解酶的产量和相应的控制策略,应用5L搅拌罐生物反应器对氮限制下(C/N=56/2.2)P.chrysosporium产木质素降解酶进行了放大研究.结果表明,在分批发酵试验中,锰过氧化物酶(MnP)和漆酶(Lac)分别在培养第6d和第7d达到峰值,其酶活随时间的变化规律与摇瓶试验基本相同;而采用氮限制液体培养基进行补料没有获得更高的酶活,因此,可以得出采用发酵液体培养基作为补料液不利于白腐真菌持续产酶.另外,在分批发酵和分批补料发酵过程中,均发现体系pH值变化与白腐真菌生长和次生代谢产酶具有较好的相关性,当白腐真菌进入次生代谢阶段开始产酶时,体系pH值开始下降,随着发酵后期酶活的降低,pH值下降的幅度也逐渐变小,当体系酶活接近为0.0U/L时,pH值基本不再变化.因此,在实际发酵过程中,可以依据体系pH的变化间接了解白腐真菌的生长和产酶情况,而分批补料策略还有待于进一步研究.     

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 l0 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 chrysospodum, 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.     

Biodegradation of polycyclic aromatic hydrocarbons (PAHs) by fungal enzymes: A review

Polycyclic aromatic hydrocarbons (PAHs) are a large group of chemicals. They represent an important concern due to their widespread distribution in the environment, their resistance to biodegradation, their potential to bioaccumulate and their harmful effects. Several pilot treatments have been implemented to prevent economic consequences and deterioration of soil and water quality. As a promising option, fungal enzymes are regarded as a powerful choice for degradation of PAHs. Phanerochaete chrysosporium, Pleurotus ostreatus and Bjerkandera adusta are most commonly used for the degradation of such compounds due to their production of ligninolytic enzymes such as lignin peroxidase, manganese peroxidase and laccase. The rate of biodegradation depends on many culture conditions, such as temperature, oxygen, accessibility of nutrients and agitated or shallow culture. Moreover, the addition of biosurfactants can strongly modify the enzyme activity. The removal of PAHs is dependent on the ionization potential. The study of the kinetics is not completely comprehended, and it becomes morem hallenging when fungi are applied for bioremediation. Degradation studies in soil are much more complicated than liquid cultures because of the heterogeneity of soil, thus, many factors should be considered when studying soil bioremediation, such as desorption and bioavailability of PAHs. Different degradation pathways can be suggested. The peroxidases are heme-containing enzymes having common catalytic cycles. One molecule of hydrogen peroxide oxidizes the resting enzyme withdrawing two electrons. Subsequently, the peroxidase is reduced back in two steps of one electron oxidation. Laccases are copper-containing oxidases. They reduce molecular oxygen to water and oxidize phenolic compounds.     

葡萄糖补料对白腐真菌P.chrysosporium产木质素降解酶的影响

为了提高木质素降解酶的产量,采用氮限制培养基(C/N=56.0/8.8),研究了葡萄糖补料对P.chrysosporium产木质素降解酶的影响.结果表明,在培养第1d即接种24h后进行葡萄糖补料,可以刺激菌丝体的生长和产酶,补料浓度达到2g·L-1时,补料体系中的MnP酶活可提高至空白对照样(不进行补料)的2.5倍.同时,对葡萄糖分批补料方式进行的试验结果显示,培养过程中每24h进行1次补料,葡萄糖补料终浓度为1.5g·L-1的补料方式,与每48h的补料方式相比,产酶效果更好.MnP酶活峰值和达到峰值时酶的总产量可提高至空白对照样(不进行补料)的2.7倍和3.0倍,且酶活能够在200U·L-1以上稳定4d.实验结果说明,葡萄糖补料可以有效刺激白腐真菌产酶,且连续低浓度葡萄糖补料可获得较优的产酶效果.     

原生质体紫外诱变提高简青霉的木质素降解性能

以简青霉Penicillium simplicissimum (Oudem.) Thom BGA为出发菌株,对其进行原生质体紫外诱变.经过筛选获得2株诱变菌株(J12与J18),它们的木质素降解酶酶活要明显高于出发菌株J.其中诱变菌株J12产生的漆酶和锰过氧化物酶酶活较高,相对于出发菌株J分别提高了145%和47%,达到44.0,50.9 U/g.诱变菌株J18产生的木质素过氧化物酶酶活较高,达到67.1 U/g,相对于出发菌株J提高了40%.且木质素降解率也最高,相对于出发菌株J分别提高了198.1%.经过7次传代, J12和J18的木质素降解酶活性保持稳定,没有降低.