粗毛栓菌菌丝球非灭菌条件下对12种染料的脱色研究

以新型白腐真菌——粗毛栓菌Trametes gallica为材料,研究了优化后的该菌菌丝球在非灭菌条件下对直接染料、中性染料、三苯甲烷类染料以及蒽醌类染料共12种染料的脱色能力、脱色机制,以及pH、温度、染料初始浓度等参数对该菌菌丝球脱色效果的影响。结果表明,优化条件下制备的粗毛栓菌菌丝球脱色活力良好,4℃下保存20d后仍保持有原脱色活力的95%;活菌丝球比死菌丝球对染料具有更强的耐受性和更好的脱色效果;非灭菌条件下活菌丝球对12种染料的适宜脱色条件为pH 3.0–5.0、25℃、染料浓度为50mg/L、处理36–60h,该条件下粗毛栓菌菌丝球在60h内脱色率均在55%以上,其中粗毛栓菌菌丝球对亚甲基蓝脱色率最高可达到96.40%。紫外可见光谱分析和显微观察结果表明,48h内粗毛栓菌菌丝球在非灭菌条件下对12种染料的脱色是由吸附引起,无二次污染物的产生。粗毛栓菌的这些优良特性显示了其在工业染料废水处理中的广阔应用前景。     

新月弯孢霉菌丝球对染料脱色作用的研究

研究了新月弯孢霉Curvularia lunata JQH-100液体培养时产生的菌丝球对多种染料的脱色能力。结果表明,多种染料在24h内的脱色率均达到80%以上,且菌丝球稳定性良好,可重复使用6次;以菌丝球对孔雀绿脱色效果为优化指标,正交实验优化获得制备菌丝球的最佳条件为:葡萄糖20g/L、硫酸铵5g/L、马铃薯200g/L、KH2PO43g/L、MgSO45mg/L、CuSO40.5mg/L、VB15mg/L及pH5、摇床转速120r/min。在上述优化后的基础培养基(不含MgSO4、CuSO4)中分别添加微量元素Cu2+、Mn2+、Mg2+或Ca2+制备的菌丝球,对孔雀绿脱色能力增强;添加Fe2+制备的菌丝球,对孔雀绿脱色能力下降;分别添加Zn2+、Al3+或Na+制备的菌丝球对孔雀绿脱色能力与对照相近。应用优化后培养条件制备的菌丝球处理含多种染料的混合废水,也获得了较好的脱色效果。     

The evaluation of pre-grown mycelial pellets in decolorization of textile dyes during repeated batch process

This study was undertaken for the possibility of application of pre-grown pellets for biotechnological treatment of dyes and textile industry waste waters. Mycelial pellets of five different white rot fungi were tested for their dye decolorization activity. The pellets of Funalia trogii, Phanerochaete chrysosporium and Trametes versicolor were determined as the most effective ones. The decolorization ability of viable pellets was compared with the decolorization (adsorption) ability of dead pellets during repeated batch studies. Astrazon Black dye was decolorized effectively, about 90%, by viable pellets of all fungi during the first use. Viable F. trogii pellets were found as the most effective pellets. Upon pellet treatment not only a high decolorization but also reduced toxicity (antimicrobial activity) of the Astrazon Black dye was recorded. This type of decolorization activity with commercial or crude laccase was partially observed. Growing cells of F. trogii in batch system showed lower efficiency in color removal of mixed dyes compared to the pre-grown pellets in repeated batch system. The results in this study showed that mycelial pellets could effectively be used as an alternative to traditional physicochemical processes.     

Degradation and decolorization of monosodium glutamate wastewater with <Emphasis Type="Italic">Coriolus versicolor</Emphasis>

Degradation and decolorization of monosodium glutamate wastewater (MSGW) with Coriolus versicolor were firstly carried out. The effects of various operation parameters namely wastewater concentrations, pH, culture time and incidence of sterilization on maximum percentage of degradation and decolorization of wastewater were investigated. Studies of mycelium and enzyme for C. versicolor degradation and decolorization were estimated in this study. Ten percentage of wastewater concentration and pH = 5.0 were found to be the most suitable ones among the other experiments. The highest degradation and decolorization efficiency of wastewater was obtained at the fifth day of cultivation, which was displayed with more than 70% chemical oxygen demand removal, 83% total sugar removal and 55% color removal, respectively. Sterile operation had no remarkable effect on the degradation and decolorization efficiency for C. versicolor. Mycelium and the extra cellular fungal enzyme were both necessary for the degradation and decolorization of MSGW. C. versicolor possesses great potential and economic advantages in MSGW treatment.     

Fungal decolorization of dye wastewaters: a review

In recent years, there has been an intensive research on fungal decolorization of dye wastewater. It is becoming a promising alternative to replace or supplement present treatment processes. This paper examines various fungi, living or dead cells, which are capable of decolorizing dye wastewaters; discusses various mechanisms involved; reports some elution and regeneration methods for fungal biomass; summarizes the present pretreatment methods for increasing the biosorption capacity of fungal biomass; discusses the effect of various factors on decolorization.     

Improvement of the decolorization of azo dye by anaerobic sludge bioaugmented with<Emphasis Type="Italic">Desulfovibrio desulfuricans</Emphasis>

A culture of anaerobic sludge was bioaugmented withDesulfovibrio desulfuricans for the color removal of authentic textile wastewater containing a substantial amount of sulfate, in order to improve the decolorization process. The sulfide produced by sulfate respiration ofD. desulfuricans can chemically reduce azo bonds to produce a colorless metabolite in the form of aromatic amines. In the case where the culture of anaerobic sludge was bioaugmented withD. desulfuricans, the decolorization of C.I. Reactive Black 5 showed an increase of more than 14% after 48 h in comparison with that in the culture of anaerobic sludge alone. In the decolorization of authentic textile wastewater, the color removal (about 69.0%) was improved by the mixed culture of anaerobic sludge andD. desulfuricans, compared with results obtained with only anaerobic sludge as reported in our previous work, suggesting that bioaugmentation byD. desulfuricans can be useful for the decolorization of wastewater that contains complex dye compounds and sulfate.     

Autocatalysis in Reactive Black 5 biodecolorization by <Emphasis Type="Italic">Rhodopseudomonas palustris</Emphasis> W1

Autocatalysis in biological decolorization of Reactive Black 5 (RB5) by Rhodopseudomonas palustris W1 was investigated in batch assays. An improvement of 1.5-fold in decolorization rate of RB5 was obtained by supplementing decolorization metabolites from 200 mg l(-1) RB5. Liquid chromatography-mass spectrometry and cyclic voltammetric analysis revealed that the constituent of dye precursors, from azo bonds breakage, with quinone-like structure and reversible oxidation-reduction activity can be used as redox mediators and was responsible for the catalytic reduction of RB5. The required amount of metabolites for catalytic decolorization was quite small, indicating its possible application in real textile wastewater treatment. Furthermore, decolorization metabolites of RB5 were shown as effective in catalyzing anaerobic decolorization of Direct Yellow 11, an azo dye without autocatalyic activity.     

Distribution of live and dead cells in pellets of an actinomycete Amycolatopsis balhimycina and its correlation with balhimycin productivity

Secondary metabolites such as antibiotics are typically produced by actinomycetes as a response to growth limiting stress conditions. Several studies have shown that secondary metabolite production is correlated with changes observed in actinomycete pellet morphology. Therefore, we investigated the correlation between the production of balhimycin and the spatio-temporal distribution of live and dead cells in pellets of Amycolatopsis balhimycina in submerged cultures. To this end, we used laser scanning confocal microscopy to analyze pellets from balhimycin producing and nonproducing media containing 0.2 and 1.0 g l^?1 of potassium di-hydrogen phosphate, respectively. We observed a substantially higher fraction of live cells in pellets from cultures yielding larger amounts of balhimycin. Moreover, in media that resulted in no balhimycin production, the pellets exhibit an initial death phase which commences from the centre of the pellet and extends in the radial direction. A second growth phase was observed in these pellets, where live mycelia are seen to appear in the dead core of the pellets. This secondary growth was absent in pellets from media producing higher amounts of balhimycin. These results suggest that distribution of live and dead cells and its correlation with antibiotic production in the non-sporulating A. balhimycina differs markedly than that observed in Streptomycetes.     

Semicontinuous decolorization of azo dyes by rotating disc contactor immobilized with<Emphasis Type="Italic">Aspergillus sojae</Emphasis> B-10

Aspergillus sojae B-10 was immobilized and used to treat model dye compounds. The model wastewater, containing 10 ppm of azo dyes such as Amaranth, Sudan III, and Congo Red, was treated with cells attached to a rotating disc contactor (RDC). Amaranth was decolorized more easily than were Sudan III and Congo Red. Decolorization of Amaranth began within a day, and the dye was completely decolorized within 5 days of incubation. Both Sudan III and Congo Red were almost completely decolorized after 5 days of incubation. Semicontinuous decolorization of azo by reusing attached mycelia resulted in almost complete decolorization in 20 days. This experiment indicated that decolorization was successfully conducted by removing azo dyes withAspergillus sojae B-10.     

Potential of combined fungal and bacterial treatment for color removal in textile wastewater

Low efficiency of dye removal by mixed bacterial communities and high rates of dye decolorization by white-rot fungi suggest a combination of both processes to be an option of treatment of textile wastewaters containing dyes and high concentrations of organics. Bacteria were able to remove mono-azo dye but not other chemically different dyes whereas decolorization rates using Irpex lacteus mostly exceeded 90% within less than one week irrespective of dye structure. Decolorization rates for industrial textile wastewaters containing 2-3 different dyes by fungal trickling filters (FTF) attained 91%, 86%, 35% within 5-12 d. Sequential two-step application of FTF and bacterial reactors resulted in efficient decolorization in 1st step (various single dyes, 94-99% within 5 d; wastewater I, 90% within 7 d) and TOC reduction of 95-97% in the two steps. Large potential of combined use of white-rot fungi and traditional bacterial treatment systems for bioremediation of textile wastewaters was demonstrated.     

绒毛栓孔菌菌丝体在无营养条件下对偶氮染料刚果红的脱色作用

【目的】在无营养条件下,利用白腐真菌绒毛栓孔菌(Trametes pubescens)菌丝体对染料进行脱色可减少试验成本,提高染料处理的实用性。【方法】将该菌株液体培养的菌丝体在无营养条件下对染料进行脱色,并对其中脱色效果较好的偶氮染料刚果红的脱色过程进行分析。在此过程中,测定了该菌株分泌的胞外胞内酶活力,优化影响因子如初始pH值、温度、染料浓度和盐度,同时利用气相色谱-质谱联用技术分析无营养条件下偶氮染料刚果红的降解产物。植物毒性试验测定刚果红经绒毛栓孔菌菌丝体脱色前后的毒性变化。【结果】菌丝体对偶氮染料刚果红有较好的脱色效果,在初始pH值为2.0,温度为30°C,染料浓度为80 mg/L,盐度为2.5%(质量体积比)时,150 r/min转速下培养7 d后脱色率可达80.52%。在此过程中,菌丝体可被连续使用2次,且其所分泌的酶系可降解染料。此外,通过气相色谱-质谱联用分析得到刚果红的降解产物为萘胺、联苯胺和叠氮萘。植物毒性试验显示在无营养条件下的绒毛栓孔菌菌丝体对染料有明显的脱毒作用。【结论】研究发现绒毛栓孔菌菌丝体在无营养条件下的偶氮染料废水处理中具有广阔的应用前景。     

Isolation and characterization of a Klebsiella oxytoca strain for simultaneous azo-dye anaerobic reduction and bio-hydrogen production

A facultative anaerobic bacteria strain GS-4-08, isolated from an anaerobic sequence batch reactor for synthetic dye wastewater treatment, was investigated for azo-dye decolorization. This bacterium was identified as a member of Klebsiella oxytoca based on Gram staining, morphology characterization and 16S rRNA gene analysis. It exhibited a good capacity of simultaneous decolorization and hydrogen production in the presence of electron donor. The hydrogen production was less affected even at a high Methyl Orange (MO) concentration of 0.5 mM, indicating a superior tolerability of this strain to MO. This efficient bio-hydrogen production from electron donor can not only avoid bacterial inhibition due to accumulation of volatile fatty acids during MO decolorization, but also can recover considerable energy from dye wastewater.     

Efficient secretion of three fungal laccases from Saccharomyces cerevisiae and their potential for decolorization of textile industry effluent—A comparative study

Laccases are enzymes with a broad range of biotechnological applications and have, for example, the ability to oxidize many xenobiotics including synthetic dyes. In order to obtain an efficient laccase for the decolorization of dyes which spoil wastewater from the textile industry, genes encoding three various laccase enzymes were expressed in Saccharomyces cerevisiae. The expression of laccases from ascomycete Myceliophthora thermophila (MtL), and two basidiomycetes Trametes versicolor (TvL) and Trametes trogii (TtL) was optimized via selection of plasmids, promoters, media composition, and cultivation conditions. For the first time, the activity of the three secreted laccases was directly compared with the use of various substrates, including different dyes and a wastewater sample. A strong constitutive ADH1 promoter, minimal growth medium, optimized combination of copper and organic nitrogen source, and low cultivation temperature were shown to significantly increase the yields and relative activities of secreted laccases. Heterologous expression of three fungal laccases was successfully achieved in S. cerevisiae being the highest for MtL and the lowest for TvL. MtL, and particularly TtL, showed the decolorization capacity. This is the first report which compared decolorization of synthetic dyes and wastewater by several recombinant laccases and suggested MtL and TtL to be applicable in the ecofriendly enzymatic treatment of colored industry effluent. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 34:69–80, 2018     

Comparison of Pb2 accumulation characteristics between live and dead cells of Saccharomyces cerevisiae and Aureobasidium pullulans

Pb2+ accumulation processes between live and dead cells of Saccharomyces cerevisiae and Aureobasidium pullulans are different. In the case of S. cerevisiae, the Pb2+ accumulation capacity of the live cells was higher than that of the dead cells but they showed reversed initial Pb2+ accumulation rates. On the contrary, A. pullulans used a different process due to the existence of extracellular polymeric substances, which allowed both the capacity and the initial rate of Pb2+ accumulation in the live cells to be higher than those in the dead cells. © Rapid Science Ltd. 1998     

Decolorization of an industrial effluent by free and immobilized cells of <Emphasis Type="Italic">Stenotrophomonas maltophilia</Emphasis> AAP56<Emphasis Type="Italic">.</Emphasis> Implementation of efficient down flow column reactor

A bacterial strain AAP56, isolated from a polluted soil (from Kelibia city) and identified as Stentrophomonas maltophilia, was particularly interesting for its ability to decolorize recalcitrant dyes of an industrial effluent: SITEX Black. The final percentage decolorization 60% was shown by bacterial culture after incubation in LB medium at 30°C under shaking conditions. The decolorization was closely correlated with the metabolic bacterial growth. The replacement of yeast extract in LB medium composition by soya flour was clearly efficient to enhance the percentage decolorization by 20% and also to reduce the growth medium cost 60-fold. The bacteria were able to reduce 23% from the initial COD and 28% from the initial BOD₅ of the effluent. The immobilization of bacterial cells in calcium alginate beads improved by 25% the effectiveness of the biotransformation within 24 h in batch conditions. The potential of a downflow fixed column reactor (DFCR) to decolorize SITEX Black was evaluated under dilution rate. The best decolorization percentage (82%) was recorded at 0.3 h⁻¹. This bioprocess seems to be a potentially useful method to remediate the colored textile wastewater.     

白囊耙齿菌对茜素红脱色条件优化及其毒性变化检测

白囊耙齿菌Irpex lacteus是分离自倒木上的一株可以分泌漆酶和锰过氧化物酶的白腐真菌。利用I. lacteus对固体条件下的活性黑、活性红、结晶紫、茜素红和孔雀石绿进行脱色能力的检测,通过单因素和正交试验优化I. lacteus对茜素红的脱色条件,并以3种作物发芽率为指标测定茜素红被I. lacteus脱色前后的毒性变化。结果显示,I. lacteus对5种染料均可脱色,其中对茜素红染料的脱色更为彻底;单因素和正交试验优化I. lacteus对茜素红的脱色条件为：pH 7.0、葡萄糖10.0g/L、硫酸铵0.66g/L、接种量2片（Φ=8.0mm）、100.0mL三角瓶装液20.0mL,优化条件下I. lacteus对茜素红脱色10d时的脱色率为88.26%,与未优化前的脱色率相比提高了60.50%;茜素红染料被I. lacteus脱色前后毒性大小排序为：染料原液>染料脱色后>PDB培养基处理,表明茜素红染料存在一定的毒性,I. lacteus脱色茜素红后可以使其毒性减弱。通过本研究,为I. lacteus在茜素红等染料废水脱色以及降低染料废水毒性方面的应用奠定基础。     

Predicting the effects of chlorine on the micro-organisms of filamentous bulking activated sludges

Rapid and definite assessment of the effect that a specific biocide has on a specific case of filamentous bulking sludge is a much-needed tool in activated sludge wastewater treatment. The Live/Dead stain (LIVE/DEAD BacLight) distinguishing "living" and "non-living" cells, a nitrifying activity (NA) test and the oxygen uptake rate (OUR) measurement were examined for their appropriateness to predict the effects of chlorine on filamentous bulking sludges. The study showed the live/dead stain to be relevant for revealing the specific effect of chlorine on the filamentous bacteria of a bulking sludge. However, using live/dead stain alone for the determination of the appropriate chlorine dose against bulking may lead to an underestimation of the damage caused by chlorine to the useful microorganisms in the flocs. Indeed, using the live/dead stain, it was not easy to distinguish dead cells caused by chlorination from those originally present in the flocs The NA test was the most sensitive in detecting damage caused by chlorine to the floc-forming microorganisms. Therefore, for a safer determination of the chlorine dose effective against bulking and protective of the microbial activity of the sludge, the results of this study suggest coupling of the live/dead stain with the NA test and/or the OUR test.     

Mass and Density Measurements of Live and Dead Gram-Negative and Gram-Positive Bacterial Populations

Monitoring cell growth and measuring physical features of food-borne pathogenic bacteria are important for better understanding the conditions under which these organisms survive and proliferate. To address this challenge, buoyant masses of live and dead Escherichia coli O157:H7 and Listeria innocua were measured using Archimedes, a commercially available suspended microchannel resonator (SMR). Cell growth was monitored with Archimedes by observing increased cell concentration and buoyant mass values of live growing bacteria. These growth data were compared to optical density measurements obtained with a Bioscreen system. We observed buoyant mass measurements with Archimedes at cell concentrations between 10⁵ and 10⁸ cells/ml, while growth was not observed with optical density measurements until the concentration was 10⁷ cells/ml. Buoyant mass measurements of live and dead cells with and without exposure to hydrogen peroxide stress were also compared; live cells generally had a larger buoyant mass than dead cells. Additionally, buoyant mass measurements were used to determine cell density and total mass for both live and dead cells. Dead E. coli cells were found to have a larger density and smaller total mass than live E. coli cells. In contrast, density was the same for both live and dead L. innocua cells, while the total mass was greater for live than for dead cells. These results contribute to the ongoing challenge to further develop existing technologies used to observe cell populations at low concentrations and to measure unique physical features of cells that may be useful for developing future diagnostics.     

壳聚糖固定化气单胞菌处理餐厨高油脂废水

为有效解决餐厨废水中的高油脂对下游处理工艺的严重影响,本研究以筛选获得的一株高效的油脂降解菌株嗜糖气单胞菌Aeromonas allosaccarophila CY-01为研究对象,以壳聚糖为载体材料,对其进行固定化包埋制备壳聚糖-气单胞菌小球(CH-CY01);探究CH-CY01小球的油脂降解效率以及性能影响评估。研究结果表明,经壳聚糖固定后的菌株CY-01其细胞的生长活性几乎不受影响,对大豆油脂的最大降解率为89.7%;相比于未固定的细胞,CH-CY01在0.5%NaCl的盐度条件下对油脂的降解效率显著提高了20%;在浓度为1 mg/L的表面活性剂(十二烷基苯磺酸钠)存在条件下,CH-CY01对油脂的降解效率显著提高了40%。以餐饮高油脂污水为处理对象,结果显示经添加1%(V/V)的CH-CY01小球处理7 d后,超过80%以上的固态油脂被降解,明显高于未固定的CY-01。综上所述,经壳聚糖固定包埋的CH-CY01小球显著提高了菌株的生存能力和油脂降解效率,对于高油脂餐饮污水的高效处理具有较大的应用潜力。     

Removal of Humic Substances from Aqueous Solution by Fungal Pellets

The removal of humic substances from aqueous solution at pH=4.8 and 6.0 and 25 °C by pure culture of Aspergillus niger has been examined. The removal of humic substances from aqueous phase was monitored by following the decrease in absorbance at 370 nm. COD and fungal growth were also measured. The initial concentration of humic and fulvic acids, pH and diameter of fungal pellets were found to be the most important parameters. The results suggested that decolorization of batch culture was a consequence of several processes. In the first 5 hours for pellets with diameter 1-3 mm and 10 hours for pellets with diameter 2-5 mm adsorption of humic substances was dominant. Afterwards, desorption of slightly bound humic substances and their degradation, as well as degradation of remaining humic substances in the solution, continued simultaneously through the period of 10 days.