降解2,4-二氯酚微生物的分离及其2,4-二氯酚羟化酶基因的克隆和表达

从土壤中分离到一株降解2,4-二氯酚能力较强的细菌菌株GT241-1,经鉴定该菌株属于假单胞菌属。菌株GT241-1在最适条件下能在48h内将90mg/L的2,4-DCP降解91%,能利用2,4-二氯酚、2,4-二氯苯氧乙酸、苯甲酸和儿茶酚为唯一碳源生长。采用Southern杂交对2,4-二氯酚羟化酶基因（dcpA）定位后构建基因组文库,再用斑点杂交筛选目的转化子,克隆了该菌株的dcpA。序列测定得知含dcpA的亚克隆片段全长2389bp,其中dcpA基因编码区1797bp。核苷酸和氨基酸序列分析表明,dcpA与已在GenBank登记的相关基因有一定的差异。dcpA基因能够在大肠杆菌转化子中成功地表达有生物活性的酶。     

白腐真菌生物技术降解氯酚污染物

生物降解是降解氯酚污染物的一条重要的转化途径,白腐真菌是一种高效的生物降解菌种,应用白腐真菌生物技术降解具有毒性和抗降解性的氯酚具有重要意义。本文阐述了白腐菌降解氯酚类污染物的途径,阐述了白腐真菌技术,主要包括酶技术、固定化技术、真菌强化技术、堆肥化和生物反应器等在氯酚污染环境治理中的应用,并概述了近几年白腐菌降解氯酚的研究热点和白腐真菌生物技术的应用趋势。     

固定化白腐真菌对多种染料脱色的研究

白腐真菌F17 Schizophyllum sp．对不同结构的多种染料具有较强的脱色能力。聚酯无纺布是该菌固定化的最佳载体。利用正交实验,确定了该菌株固定化细胞制备的最优操作条件。与游离菌相比,固定化菌不仅提高其脱色能力,而且在pH值和染料浓度的变化情况下,仍保持稳定的脱色率。固定化菌对染料脱色后的紫外可见光谱分析表明,可见光区吸收峰消失,紫外区的光吸收峰有所增加,染料结构发生了变化。     

对污染物有独特降解作用的白腐真菌

简述了白腐真菌对木质素的特殊降解过程,进而讨论了该真菌在降解污染物及造纸制浆方面可能的潜在用途。     

白腐真菌对酸性橙7的脱色降解

偶氮染料广泛应用于纺织、造纸和包装等行业,因其具有三致性、结构稳定且难降解,已成为染料废水处理的研究热点之一。本研究以白腐真菌作为脱色菌株,考察了不同白腐真菌对偶氮类染料酸性橙7(acid orange 7,AO7)的脱色降解,探讨了AO7染料的浓度、pH、温度以及脱色时间对染料脱色率的影响,同时应用紫外-可见光谱吸收法、红外光谱吸收法、高效液相色谱法和气相色谱-质谱法对AO7的降解产物进行分析,并对其产物进行植物毒性实验,以推断AO7可能的降解途径及其降解产物的毒性。结果表明:在pH 4.5、28℃条件下,刺芹侧耳(Pleurotus eryngii)和杂色云芝(Trametes versicolor)的混合菌丝脱色降解100 mg/L AO7,24 h脱色率可达93.46%。推测AO7可能的生物降解途径:AO7偶氮键断裂生成对氨基苯磺酸和1-氨基-2-萘酚;接着对氨基苯磺酸脱去磺酸基,生成对苯二酚;同时1-氨基-2-萘酚开环生成邻苯二甲酸和对羟基苯甲醛,之后进一步降解生成苯甲酸;最后对苯二酚和苯甲酸继续氧化成其他小分子中间体、H2O和CO_(2)。植物毒性实验表明,P.eryngii和T.versicolor混合菌丝对AO7脱毒效果较好。以上研究为探究白腐真菌在工业废水中降解偶氮类染料的应用奠定基础。     

白腐真菌及其技术的潜在工业应用

本文概述了白腐真菌对木质素和各种结构不同的化学物的独特降解能力及其在降解机制,以及白腐真菌技术在石油化学工业,煤炭工业,纸浆造纸工业,纺织印染工业,酶制品生产和环境工程等方面潜在的工业应用前景。     

固定化白腐真菌对多种染料脱色的研究*

白腐真菌F17Schizophyllumsp.对不同结构的多种染料具有较强的脱色能力。聚酯无纺布是该菌固定化的最佳载体。利用正交实验,确定了该菌株固定化细胞制备的最优操作条件。与游离菌相比,固定化菌不仅提高其脱色能力,而且在pH值和染料浓度的变化情况下,仍保持稳定的脱色率。固定化菌对染料脱色后的紫外可见光谱分析表明,可见光区吸收峰消失,紫外区的光吸收峰有所增加,染料结构发生了变化。     

白腐真菌对染料废水脱色及降解的研究

染料废水是最难处理的工业废水之一,近年来许多学者就白腐真菌对染料废水的脱色进行了广泛的研究,系统介绍了白腐真菌对染料脱色和降解作用的研究进展,脱色机理及其影响因素,旨在为以后真菌对染料废水的脱色及降解提供参考和依据。     

混合白腐真菌的固定化及其在治理铅污染废水中的应用

【目的】为缓解重金属废水污染对全球食品安全和人类健康的威胁,降低铅(plumbum, Pb)在土壤及动植物体内的积累,借助固定化技术提高菌株的重金属去除效率。【方法】以白腐真菌(white rot fungi)为实验材料,通过混菌兼容性及铅离子(Pb²⁺)去除能力筛选出吸附效果好且兼容性优的复合菌种,探究最优混菌类型及其比例,优化菌球最佳固定化助剂配方,在此基础上深入探究菌球在实际应用中的最优吸附条件。【结果】黄孢原毛平革菌(Phanerochaete chrysosporium)、云芝(Coriolus versicolor)、凤尾菇(Lentinus sajor-caju)和平菇(Pleurotus ostreatus) 4种菌株兼容效果佳,可进行后续实验;其中云芝和凤尾菇以体积1:1混合后对Pb²⁺去除效果显著优于各单菌作用;固定化条件优化实验中,20.0 g/L海藻酸钠、15.0 g/L生物炭和2.0×10⁶个/mL白腐真菌组成混菌体系,辅以二氧化硅及沸石制得的固定化菌球在96 h Pb²⁺...     

白腐真菌的木质素降解酶

简述了白腐真菌木质素降解酶的概念、催化反应机理及在纸浆的生物漂白和染料脱色中的应用。     

Biodegradation of polycyclic hydrocarbons by Phanerochaete chrysosporium

The ability of the white rot fungus Phanerochaete chrysosporium to degrade polycyclic aromatic hydrocarbons (PAHs) that are present in anthracene oil (a distillation product obtained from coal tar) was demonstrated. Analysis by capillary gas chromatography and high-performance liquid chromatography showed that at least 22 PAHs, including all of the most abundant PAH components present in anthracene oil, underwent 70 to 100% disappearance during 27 days of incubation with nutrient nitrogen-limited cultures of this fungus. Because phenanthrene is the most abundant PAH present in anthracene oil, this PAH was selected for further study. In experiments in which [14C]phenanthrene was incubated with cultures of P. chrysosporium containing anthracene oil for 27 days, it was shown that 7.7% of the recovered radiolabeled carbon originally present in [14C]phenanthrene was metabolized to 14CO2 and 25.2% was recovered from the aqueous fraction, while 56.1 and 11.0% were recovered from the methylene chloride and particulate fractions, respectively. High-performance liquid chromatography of the 14C-labeled material present in the methylene chloride fraction revealed that most (91.9%) of this material was composed of polar metabolites of [14C]phenanthrene. These results suggest that this microorganism may be useful for the decontamination of sites in the environment contaminated with PAHs.     

Biodegradation of Rose Bengal by Phanerochaete chrysosporium

Rose Bengal (tetrachloro-tetraiodo-fluorescein) was not able to limit the spreading growth of the ligninolytic fungus, Phanerochaete chrysosporium in the presence of Tween 80, and when added to the 5 d old liquid cultures of this organism it was almost completely degraded in 5 h. Thin layer chromatography analysis showed the formation of a single degradation product.     

Biodegradation of polycyclic hydrocarbons by Phanerochaete chrysosporium.

The ability of the white rot fungus Phanerochaete chrysosporium to degrade polycyclic aromatic hydrocarbons (PAHs) that are present in anthracene oil (a distillation product obtained from coal tar) was demonstrated. Analysis by capillary gas chromatography and high-performance liquid chromatography showed that at least 22 PAHs, including all of the most abundant PAH components present in anthracene oil, underwent 70 to 100% disappearance during 27 days of incubation with nutrient nitrogen-limited cultures of this fungus. Because phenanthrene is the most abundant PAH present in anthracene oil, this PAH was selected for further study. In experiments in which [14C]phenanthrene was incubated with cultures of P. chrysosporium containing anthracene oil for 27 days, it was shown that 7.7% of the recovered radiolabeled carbon originally present in [14C]phenanthrene was metabolized to 14CO2 and 25.2% was recovered from the aqueous fraction, while 56.1 and 11.0% were recovered from the methylene chloride and particulate fractions, respectively. High-performance liquid chromatography of the 14C-labeled material present in the methylene chloride fraction revealed that most (91.9%) of this material was composed of polar metabolites of [14C]phenanthrene. These results suggest that this microorganism may be useful for the decontamination of sites in the environment contaminated with PAHs.     

Biodegradation of bioaccessible textile azo dyes by Phanerochaete chrysosporium

Azo dyes are important chemical pollutants of industrial origin. Textile azo dyes with bioaccessible groups for lignin degrading fungi, such as 2-methoxyphenol (guaiacol) and 2,6-dimethoxyphenol (syringol), were synthesised using different aminobenzoic and aminosulphonic acids as diazo components. The inocula of the best biodegradation assays were obtained from a pre-growth medium (PAM), containing one of the synthesised dyes. The results of the dye biodegradation assays were evaluated every 7 days, by the decrease of the absorbance at the maximum wavelength of the dye, by the decrease of the sucrose concentration in the culture medium and by the increase of the biomass during the 28 days of assay. It was observed that the extent of dye biodegradation depended on the sucrose concentration, on the degraded dye structure and, on the dye present in the PAM medium.     

Biodegradation of azo and heterocyclic dyes by Phanerochaete chrysosporium

Biodegradation of Orange II, Tropaeolin O, Congo Red, and Azure B in cultures of the white rot fungus, Phanerochaete chrysosporium, was demonstrated by decolarization of the culture medium, the extent of which was determined by monitoring the decrease in absorbance at or near the wavelength maximum for each dye. Metabolite formation was also monitored. Decolorization of these dyes was most extensive in ligninolytic cultures, but substantial decolorization also occurred in nonligninolytic cultures. Incubation with crude lignin peroxidase resulted in decolorization of Azure B, Orange II, and Tropaeolin O but not Congo Red, indicating that lignin peroxidase is not required in the initial step of Congo Red degradation.     

Biodegradation and sorption of polyaromatic hydrocarbons by Phanerochaete chrysosporium

The ability of the white-rot fungus Phanerochaete chrysosporium (INA-12) to degrade various polynuclear aromatic hydrocarbons (PAH) was investigated. Under static, non-nitrogen-limiting conditions, P. chrysosporium mineralized both phenanthrene and benzo[a]pyrene. Total mineralization, based on radioactive tracing, was limited to 1.8%–3% for phenanthrene and benzo[a]pyrene respectively. In both cases the pattern of mineralization did not correlate temporally with the production of lignin peroxidase activity. Sorption of radiolabelled material to the biomass was very significant with 22% and 40% of the total radioactivity being sorbed for benzo[a]pyrene and phenanthrene respectively. A number of models were examined to predict the sorption isotherms, the best performance being obtained with a three-parameter empirical model. It is apparent that lignin peroxidase is not necessarily involved in the biodegradation of all PAH and that a significant factor in PAH biodegradation and/or disappearance in cultures with the intact fungus may be attributed to sorption phenomena.     

Biodegradation of TNT (2,4,6-trinitrotoluene) by Phanerochaete chrysosporium

Extensive biodegradation of TNT (2,4,6-trinitrotoluene) by the white rot fungus Phanerochaete chrysosporium was observed. At an initial concentration of 1.3 mg/liter, 35.4 +/- 3.6% of the [14C]TNT was degraded to 14CO2 in 18 days. The addition of glucose 12 days after the addition of TNT did not stimulate mineralization, and, after 18 days of incubation with TNT only, about 3.3% of the initial TNT could be recovered. Mineralization of [14C]TNT adsorbed on soil was also examined. Ground corncobs served as the nutrient for slow but sustained degradation of [14C]TNT to 14CO2 such that 6.3 +/- 0.6% of the [14C]TNT initially present was converted to 14CO2 during the 30-day incubation period. Mass balance analysis of liquid cultures and of soil-corncob cultures revealed that polar [14C]TNT metabolites are formed in both systems, and high-performance liquid chromatography analyses revealed that less than 5% of the radioactivity remained as undegraded [14C]TNT following incubation with the fungus in soil or liquid cultures. When the concentration of TNT in cultures (both liquid and soil) was adjusted to contamination levels that might be found in the environment, i.e., 10,000 mg/kg in soil and 100 mg/liter in water, mineralization studies showed that 18.4 +/- 2.9% and 19.6 +/- 3.5% of the initial TNT was converted to 14CO2 in 90 days in soil and liquid cultures, respectively. In both cases (90 days in water at 100 mg/liter and in soil at 10,000 mg/kg) approximately 85% of the TNT was degraded. These results suggest that this fungus may be useful for the decontamination of sites in the environment contaminated with TNT.     

Biodegradation of pentachlorophenol by the white rot fungus Phanerochaete chrysosporium

Extensive biodegradation of pentachlorophenol (PCP) by the white rot fungus Phanerochaete chrysosporium was demonstrated by the disappearance and mineralization of [14C]PCP in nutrient nitrogen-limited culture. Mass balance analyses demonstrated the formation of water-soluble metabolites of [14C]PCP during degradation. Involvement of the lignin-degrading system of this fungus was suggested by the fact the time of onset, time course, and eventual decline in the rate of PCP mineralization were similar to those observed for [14C]lignin degradation. Also, a purified ligninase was shown to be able to catalyze the initial oxidation of PCP. Although biodegradation of PCP was decreased in nutrient nitrogen-sufficient (i.e., nonligninolytic) cultures of P. chrysosporium, substantial biodegradation of PCP did occur, suggesting that in addition to the lignin-degrading system, another degradation system may also be responsible for some of the PCP degradation observed. Toxicity studies showed that PCP concentrations above 4 mg/liter (15 microM) prevented growth when fungal cultures were initiated by inoculation with spores. The lethal effects of PCP could, however, be circumvented by allowing the fungus to establish a mycelial mat before adding PCP. With this procedure, the fungus was able to grow and mineralize [14C]PCP at concentrations as high as 500 mg/liter (1.9 mM).     

Biodegradation of azo and heterocyclic dyes by Phanerochaete chrysosporium.

Biodegradation of Orange II, Tropaeolin O, Congo Red, and Azure B in cultures of the white rot fungus, Phanerochaete chrysosporium, was demonstrated by decolarization of the culture medium, the extent of which was determined by monitoring the decrease in absorbance at or near the wavelength maximum for each dye. Metabolite formation was also monitored. Decolorization of these dyes was most extensive in ligninolytic cultures, but substantial decolorization also occurred in nonligninolytic cultures. Incubation with crude lignin peroxidase resulted in decolorization of Azure B, Orange II, and Tropaeolin O but not Congo Red, indicating that lignin peroxidase is not required in the initial step of Congo Red degradation.