不同共代谢基质对白腐菌降解吲哚的作用研究

选用氨氮、苯酚和喹啉作为吲哚的共代谢基质,通过白腐菌BP对共基质体系的降解研究了白腐菌在秸秆滤出液培养基中对不同共基质体系的代谢过程,并进行了动力学分析,考察了不同共代谢基质物质对白腐菌漆酶分泌和吲哚降解的影响.结果显示,不同降解体系中的白腐菌都可去除99%以上的吲哚.充分的氮源可提高白腐菌的活性和漆酶酶活的峰值;共基质苯酚和喹啉可以增加白腐菌漆酶产量,为吲哚的降解提供较多的电子,同时苯酚和喹啉也能得到较高的去除.在秸秆滤出液中,白腐菌在pH为6～8之间对吲哚都具有较强的降解能力.吲哚在白腐菌的代谢过程中,可能首先在吡啶环的2和3位发生一步羰基化.     

白腐菌对焦化废水中喹啉的降解及机理研究

选用白腐菌BP降解喹啉,研究了白腐菌在不同培养基中对喹啉的降解过程和机理,以及喹啉的降解与白腐菌漆酶活力、生物量比增长速率、培养基pH值的关系。结果显示秸秆滤出液培养基中生长的白腐菌对喹啉具有最高的去除率89%;喹啉的相对去除率与白腐菌漆酶活力、生物量比增长速率具有较好的相关性;白腐菌BP最适pH为6~7;2-羟基喹啉是喹啉降解过程中首先出现的中间产物。     

稻草固体介质中白腐菌对喹啉的降解研究

研究了稻草固体介质中侧耳属白腐菌BP对喹啉的降解。实验结果表明,浓度分别为250mg/L和150mg/L喹啉的降解率能达到93.47%和97.40%。白腐菌BP可以有效对喹啉进行降解。漆酶和锰过氧化物酶在降解过程中起着较重要的作用,但酶活的高低与降解率大小并不完全成正比。高效液相色谱分析表明降解过程中没有中间代谢物的积累,喹啉能得到彻底矿化。     

重金属对白腐菌降解十溴联苯醚的影响

考察了重金属(Cu、Cd、Pb)对白腐菌生长及其降解十溴联苯醚(BDE-209)的影响及其机制.结果表明,低浓度重金属(≤1 mg.L-1)可促进白腐菌的生长,高浓度则表现为抑制作用,1 mg.L-1时促进作用大小为:Cd>Pb>Cu;白腐菌可高效降解BDE-209,7 d内对BDE-209(1 mg.L-1)降解率可达69.7%;重金属可显著影响白腐菌对BDE-209的降解(P<0.05),低浓度的Cu(≤1 mg.L-1)和Cd(≤0.5 mg.L-1)对BDE-209的降解表现为促进作用,其中Cu为1 mg.L-1时促进作用最明显,降解率为84.4%,而Pb则为抑制作用;高浓度(>1 mg.L-1)的重金属均会抑制BDE-209的降解,抑制作用大小为Cd>Pb>Cu,且随浓度增大抑制作用增强;降解率与生物量变化不完全正相关.白腐菌对BDE-209的降解过程符合一级反应动力学方程,Cu、Cd存在条件下,随着其浓度的增加,降解速率常数k表现为先增加后减少,Cu为1 mg.L-1时k值达到最大,为0.321 2;Pb存在条件下,k值表现为逐渐减小.进一步研究了重金属对白腐菌胞外酶降解BDE-209的影响,并运用SPSS 17.0对胞外酶降解与菌体降解进行距离相关性分析,结果表明,未添加重金属时胞外酶降解与菌体降解差异不大,其降解率分别为63.7%、69.7%;3种重金属存在条件下其相关系数R值均大于0.9,由此推断胞外酶是起降解作用的主要部分,重金属主要通过影响白腐菌胞外酶的方式作用于BDE-209的降解.     

非灭菌条件下酸性蓝45在白腐真菌反应器中的降解特性

在非灭菌条件下维持白腐真菌反应器连续运行是在水处理领域实际应用白腐真菌的前提.采用臭氧控制白腐真菌反应器中的污染细菌,以连续运行方式考察了自由悬浮培养的白腐真菌体系中控菌、产酶、pH变化及对酸性蓝45的降解特性.结果表明,在白腐真菌反应器系统的控菌单元,以0.0144 mg/min臭氧投加速率,可成功将白腐真菌反应器内污染细菌控制在1×106 CFU/mL以下,其去除率接近99%;成功实现了白腐真菌反应器连续合成锰过氧化物酶（最大剩余酶活50 U/L）和对酸性蓝45 的持续降解（40％～80％）;发现pH值在6左右时,白腐真菌培养体系仍然具有合成锰过氧化物酶和对酸性蓝45的持续降解的能力;提出在控制污染细菌的前提下,如何维持白腐真菌的持续生长及高产酶是进一步实现白腐真菌反应器连续运行的主要问题.     

蜜环菌漆酶对氯酚类污染物催化降解条件优化

利用蜜环菌发酵所得的漆酶粗酶液直接对2种氯酚类污染物2,4-二氯酚(2,4-DCP)和2-氯酚(2-CP)进行催化降解实验,探讨了反应时间、pH值、反应温度、氯酚浓度、以及漆酶酶量对其降解效果的影响,得出了最适降解条件并对其降解动力学进行分析.结果表明,在适宜的条件下,漆酶粗酶液可有效降解2,4-DCP和2-CP且蜜环菌漆酶催化降解2,4-DCP的能力较强,2,4-DCP最适降解温度为40℃,最适浓度为75 mg.L-1,最适酶量为0.1 U.mL-1,最适pH值为6.5,在最优条件下反应10h后,2,4-DCP最高降解率可达97%以上.2-CP最适降解温度为50℃,最适浓度为100 mg.L-1,最适酶量为0.1 U.mL-1,最适pH值为6,在最优条件下反应10 h后,2-CP最高降解率高达93%以上.蜜环菌漆酶粗酶液对2,4-DCP和2-CP的降解反应符合一级动力学特征.结果表明蜜环菌粗漆酶液能有效转化氯酚类化合物,说明该酶在酚类污染物治理和环境保护等方面有潜在应用价值.     

不同白腐真菌复配方式对产酶的影响

通过对1株和由多株白腐真菌复配共同参与下的复合菌产酶量和达到最高酶活所需时间进行比较,筛选出白腐真菌最佳产酶的复配方式.结果表明,青顶拟多孔菌和糙皮侧耳菌两株菌相互发生协同作用,促进漆酶的分泌,产酶量为50.45U·mL-1;青顶拟多孔菌、糙皮侧耳菌和偏肿拟栓菌这3株菌在分泌过程中促进作用大于抑制作用,产酶量最高为75.98U·mL-1;青顶拟多孔菌、糙皮侧耳菌和血红密孔菌三者相互之间出现拮抗作用,产酶量最低为2.91 U·mL-1;实验证明,在白腐真菌之间既存在拮抗作用,也存在协同作用,因此达到最高酶活所用时间也不同.     

处理秸秆纤维素乙醇废水的东北土著白腐真菌筛选及特性研究

为了提高秸秆纤维素乙醇废水的处理效果,选择6种东北土著白腐真菌,对2%的秸秆纤维素乙醇废水中木质素进行降解处理.采用正交试验法对筛选出的高效降解菌进行产漆酶培养基的优化.结果表明:6种白腐真菌最高酶活大小顺序为青顶拟多孔菌>血红密孔菌>糙皮侧耳菌>彩绒革盖菌>烟色烟管菌>灵芝;血红密孔菌、糙皮侧耳菌、彩绒革盖菌、青顶拟多孔菌、灵芝、烟色烟管菌在第0天起始质量浓度为640.9~716.6 mg/L,在第14天木质素的质量浓度分别为434.0、411.2、441.8、441.7、533.3、503.5 mg/L,对木质素的去除率分别为37.1%、37.0%、31.8%、31.7%、25.6%、21.4%,并分别在第12、12、4、4、2、6天木质素降解趋于平稳,表明降解效果最好的菌种为血红密孔菌.血红密孔菌产漆酶培养基最优组合方案:最佳碳源为锯末,质量浓度为35 g/L;最佳氮源为蛋白胨,质量浓度为4 g/L;最佳pH为5.极差分析表明,各因素对血红密孔菌产漆酶的影响顺序为碳源>氮源> pH >氮源质量浓度>碳源质量浓度.在最佳培养基条件下,废水中木质素降解率达41.1%.研究显示,血红密孔菌可以作为生物法处理秸秆纤维素乙醇废水的菌种资源,也可为今后的进一步应用研究提供科学依据.      

白腐菌对芳香化合物的降解及其机制

本文讨论了白腐菌（Ｗｈｉｔｅｒｏｔｆｕｎｇｉ）对芳香化合物的降解及其机制。白腐菌是目前发现的对芳香化合物有很强降解能力的真菌，它通过分泌木素过氧化物酶（Ｌｉｇｎｉｎｐｅｒｏｘｉｄａｓｅ）、锰过氧化物酶（Ｍａｎｇａｎｅｓｅ－ｄｅ－ｐｅｎｄｅｎｔｐｅｒｏｘｉｄａｓｅ）以单电子氧化、共代谢及脂肪过氧化等途径进行降解活动。白腐菌对芳香化合物的降解在污染的原位修复（ｉｎｓｉｔｕｂｉｏｒｅｍｅｄｉａｔｉｏｎ）上有重要的意义     

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.     

铸造废砂的环境毒性研究

采用毒性特性浸出程序、顶空气相色谱技术检测了5种铸造废砂中的重金属与有机污染物,采用发光细菌毒性检测技术和土壤酶活性实验研究了废砂的生物效应.结果表明,由于浇注金属与造型材料的不同,5种铸造废砂中的金属与有机污染物成分和浓度均有较大差异.其中,铸铁与铸钢废砂浸出液中的铁离子,铸铝废砂中的As离子超出了《地表水环境标准》规定的最大允许检出浓度.5种废砂中的有机污染物种类十分复杂,对发光细菌表现出不同程度的抑菌活性,发光强度的抑制率在30%～95%之间.此外,土壤酶活性实验结果表明,废砂中的金属污染物可能会抑制土壤中微生物的活性,而有机物则对土壤微生物的活性有一定的促进作用.因此,铸造废砂的随意排放将会对环境造成较严重的威胁.     

A win-win technique of stabilizing sand dune and purifying paper mill black-liquor

The principle and technique were reported here to produce lignin-based sand stabilizing material (LSSM) using extracted lignin from black liquor of straw paper mills. Field tests using LSSM to stabilize and green sand dunes were started in 2002. The field experiment was carried out in August 2005 when the newly formed plant community was 3 years old. The results from the comprehensive field experiment demonstrated that unlike polyvinyl acetate or foamed asphalt commonly used for dune stabilization, LSSM was plantfriendly material and could be used in combination with seeding and planting of desert species. With the help of LSSM, the desert species (i.e., Agriophyllum squarrosum (L.) Moq. and Artemisia desertorum Spreng. etc.) could be used to form community in 2–3 yeas and to stabilize sand dune e ectively. The newly formed community was sustainable under an extremely dry climate condition. The organic matter and total nitrogen in the soil increased significantly as the community were formed, while the change in P and K contents in the soil was negligible.     

Isolation of a salt tolerant laccase secreting strain of Trichoderma sp. NFCCI-2745 and optimization of culture conditions and assessing its effectiveness in treating saline phenolic effluents

Most of the hazardous pollutants are phenolic in nature and persists in the environment. The ability of laccases to oxidize phenolic compounds and reduce molecular oxygen to water has led to intensive studies of these enzymes. Therefore the fungal strains with high laccase activity and substrate affinity that can tolerate harsh environmental conditions have a potential for biotechnological applications. Salt tolerant laccase secreting fungi can be utilized in treatment of saline and phenolic rich industrial effluents such as coir effluent and textile effluent that needed to be diluted several fold before microbial treatment. This is the first study describing the isolation and optimization of a salt tolerant strain of Trichoderma sp. potential for industrial applications. The fungus was identified based on morphological characteristics and was subsequently confirmed with molecular techniques and deposited at National Fungal Culture Collections of India （NFCCI） under the Accession No. Trichoderma viride NFCCI 2745. In contrast to other laccase secreting fungi, light conditions did not exert much influence on laccase production of this strain and salinity enhanced its laccase secretion. The fungus effectively removed the phenolic content of the textile effluent, coir-ret liquor and wood processing effluent within 96 hr of incubation. The tolerance of the fungus to high salinity and phenolic compounds makes this strain ideal for treating saline and phenolic rich industrial effluents.     

Cagelike mesoporous silica encapsulated with microcapsules for immobilized laccase and 2, 4-DCP degradation

In this study, cage-like mesoporous silica was used as the carrier to immobilize laccase by a physical approach, followed by encapsulating with chitosan/alginate microcapsule membranes to form microcapsules of immobilized laccase based on layer-by-layer technology. The relationship between laccase activity recovery/leakage rate and the coating thickness was simultaneously investigated. Because the microcapsule layers have a substantial network of pores, they act as semipermeable membranes, while the laccase immobilized inside the microcapsules acts as a processing plant for degradation of 2,4-dichlorophenol. The microcapsules of immobilized laccase were able to degrade 2,4-dichlorophenol within a wide range of 2,4-dichlorophenol concentration, temperature and pH, with mean degradation rate around 62%. Under the optimal conditions, the thermal stability and reusability of immobilized laccase were shown to be improved significantly, as the removal rate and degradation rate remained over 40.2% and 33.8% respectively after 6 cycles of operation. Using mass spectrometry (MS) and nuclear magnetic resonance (NMR), diisobutyl phthalate and dibutyl phthalate were identified as the products of 2,4-dichlorophenol degradation by the microcapsules of immobilized laccase and laccase immobilized by a physical approach, respectively, further demonstrating the degradation mechanism of 2,4-dichlorophenol by microcapsule-immobilized laccase.     

静电纺丝纤维膜固定化漆酶对水中双酚A的降解性能

采用乳液静电纺丝技术原位固定漆酶,利用扫描电子显微镜(SEM)和激光共聚焦扫描显微镜(LCSM)对载漆酶电纺纤维膜的形貌结构进行表征.载酶电纺纤维呈壳-核结构,平均直径为(650±30)nm,表面有许多纳米级孔道;漆酶被成功包埋固定于纤维内部,且保留了79.8%的酶催化活性.以双酚A(Bisphenol A,BPA)为目标污染物,对比电纺纤维膜固定化漆酶和游离漆酶对BPA的降解性能,考察p H值和温度等因素对BPA降解效率的影响.研究结果表明:在优化条件下,电纺纤维膜固定化漆酶对BPA的降解率达80%左右;与游离漆酶相比,固定化漆酶对p H值和温度均表现出更好的耐受性.这主要是因为漆酶被包埋在纤维内部,纤维的聚合物外壳结构可以保护漆酶,减少外界环境对漆酶的影响,而纤维表面的多孔结构可以为漆酶与BPA的接触反应提供通道.     

Production of laccase by Coriolus versicolor and its application in decolorization of dyestuffs : (Ⅰ) Production of laccase by batch and repeated-batch processes

The production of laccase by Coriolus versicolor was studied.The effect of cultivation on laccase production by Coriolus versicolor was examined to obtain optimal medium and cultivation conditions.Both batch and repeated-batch processes were performed for laccase production.In repeated-batch fermentation with self-immobilized mycelia, total of 14 cycles were performed with laccase activity in the range between 3.4 and 14.8U/ml.     

Production of laccase by Coriolus versicolor and its application in decolorization of dyestuffs: (Ⅰ) Production of laccase by batch and repeated-batch processes

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活性滤料滤池气提系统影响因素研究

为了研究气量、水深和提砂管规格对活性滤料滤池提砂系统性能的影响,本实验以石英砂作为活性滤料滤池的提升介质,在恒定水头状态下,测量了不同管径下被提升的水和砂的体积和质量随压缩空气流量的变化.结果表明:随着气量的增大,提升系统的提砂量和提砂效率均呈先增大后减小的趋势;提砂管管径是最大提砂量的限制因素,但提升的砂水体积比与管径无关;相同气量条件下,单位空气提砂量与水深呈线性关系.因此,在活性滤料滤池提砂系统的设计应用过程中应选择合适的提砂管径,尽量增加浸水率,并注意控制气量以提升系统能效.     

Effects of nonionic surfactant Triton X-100 on the laccase-catalyzed conversion of bisphenol A

The laccase-catalyzed conversion of bisphenol A (BPA) in aqueous solutions was studied in the absence and presence of nonionic surfactant Triton X-100. It was found that the addition of Triton X-100 into the reaction system increased the conversion of BPA, especially near the critical micelle concentration of Triton X-100. Also it was found that the stability of laccase was greatly improved in the presence of TritonX-100. Studies on the endogenous fluorescence emission of laccase indicated that there existed an interaction between Triton X-100 and laccase, which was beneficial to folding and stabilizating of laccase. The binding of Triton X-100 to the laccase surface also mitigated the inactivation e ect caused by the free radicals and polymerization products. Under otherwise identical conditions, a lower dosage of laccase was needed for the higher conversion of BPA in the presence of Triton X-100.     

白腐菌漆酶催化聚合造纸废水中木素的研究

为了证实漆酶对从造纸厂二沉池出水提取出的木素的催化聚合作用,研究了白腐菌采绒革盖菌Coriolus versicolor漆酶对木素聚合的影响。在有氧条件下,通过添加漆酶和少量ABTS介体到水样中,用紫外分光光度计测定了其中木素浓度变化,利用凝胶色谱法分析了酶催化聚合木素前后的分子量的变化。结果表明:酶处理6h以后,废水中木素浓度从93.1mg/L下降到17.2mg/L。酶处理2h以后,从造纸厂污水分离的木素的分子量从31251上升到58610。造纸废水中木素及其衍生物被聚合后通过絮凝沉淀除去,从而实现废水色度与COD降低,进而为造纸废水回用提供可能。