反胶束漆酶/纤维素酶体系中油酸酯化的优化

漆酶和纤维素酶在反胶束水核中心具有较强的催化活性。为了对反胶束酶体系中油酸酯化反应深入研究,采用生物表面活性剂鼠李糖脂构建反胶束体系作为漆酶与纤维素酶酯化油酸的催化反应场所。通过实验研究了反胶束体系的不同条件对酯化产物含量的影响。对于反胶束漆酶体系,最佳酯化条件为：含水量W0 40%,鼠李糖脂临界胶束浓度20 mmol·L-1,pH值4,温度40 ℃;对于反胶束纤维素酶体系,最佳酯化条件为：含水量20%,鼠李糖脂临界胶束浓度80 mmol·L-1,pH值4,温度30 ℃。综合考虑几个因素,漆酶比纤维素酶更适合应用于反胶束中油酸的酯化。研究同时采用荧光光谱法对漆酶和纤维素酶在反胶束体系中结构性能的变化进行研究,结果表明,当反胶束体系处于最佳酯化反应条件时,荧光强度最高。     

共基质体系下漆酶介体系统降解氮杂环化合物的研究

采用2,2-连氮-双-(3-乙基苯并噻吡咯啉-6-磺酸)(ABTS)作为介体,与纯漆酶组成漆酶介体系统(LMS),分别研究了单基质和共基质体系下LMS对氮杂环化合物的降解。结果表明:(1)LMS对单基质和共基质体系中的吲哚的降解反应很快达到平衡,约为1h。(2)共基质体系中吡啶的引入,对吲哚的降解没有明显的促进或抑制作用;共基质体系中喹啉的引入,对吲哚的降解有较小的促进作用,而同时引入吡啶和喹啉,一定程度上也促进了吲哚的降解。(3)喹啉不能被LMS所降解。共基质体系中吲哚的引入,对吡啶的降解有一定的抑制作用;对喹啉的降解有一定的促进作用。(4)在LMS的作用下,推断吲哚生成的中间产物是氧化吲哚和靛红;吡啶生成的产物是戊二醛和戊二酸;喹啉最先生成2-羟基喹啉。     

漆酶-介体系统对多种不同结构染料的脱色效果

为研究漆酶在染料废水处理方面的潜力,选用毕赤酵母表达的担子菌Moniliophthora roreri来源的漆酶Mrl2,与小分子介体(1-羟基苯并三唑、藜芦醇、对香豆酸、N-羟基邻苯二甲酰亚胺) 协同,用于对5类不同结构和类型的染料(偶氮类、三苯甲烷类、蒽醌类、靛蓝类、吩噻嗪类)的脱色效果研究。结果表明：在小分子介体1-羟基苯并三唑(HOBT)的帮助下,漆酶对属于偶氮类、三苯甲烷类、蒽醌类、靛蓝类的9种染料脱色率均能达到95%以上;对结构更复杂的杂环吩噻嗪类染料天青I的脱色效率均较低,在最佳介体对香豆酸辅助下可达40%的脱色率。进一步对天青I染料脱色体系优化发现,在pH为6.0、对香豆酸浓度为2 mmol·L⁻¹、漆酶浓度为125 U·L⁻¹时,Mrl2对天青I的脱色率可达到81%。以上结果表明该重组漆酶在染料废水处理和环境保护等方面具有较好的应用价值和前景。     

漆酶/HBT介质系统对靛蓝染料及废水脱色的初步研究

以云芝(Trametes versicolor)1126发酵所得漆酶粗酶液与1-羟基苯并三唑(HBT)组成的漆酶/HBT介质系统对靛蓝染料进行脱色实验,分别考察了温度、转速、pH和HBT与漆酶的加入量等条件对靛蓝染料脱色的影响.最终确定的优化脱色条件为:温度60℃,转速200r/min, pH4.5, 100mL靛蓝染料"溶液"中粗酶液和HBT溶液分别加入2mL.以上述脱色条件对靛蓝印染废水进行脱色实验,反应80min,脱色率可达90.1%.     

真菌漆酶在树脂D380上的固定及固定化漆酶的性质

为寻求一种廉价高效的漆酶固定化方法,以树脂D380为载体、戊二醛为交联剂,结合吸附法和交联法固定真菌漆酶,并研究固定化漆酶的性质。最优固定化条件：1g载体在20mL0．7％的戊二醛溶液中交联2h后,加入2mL稀释酶液和5mLpH为4的醋酸一醋酸钠缓冲溶液,在25℃下吸附反应6h,最终获得的酶活回收率为65．94％。固定化漆酶的最适温度为20℃,最适pH为4,在4℃下保存14d仍保持约80％的酶活力。自由漆酶和固定化漆酶的Km值分别为1．207mmol／L和1．616mmol／L,说明固定后漆酶对底物的亲和力有所下降。     

石油污染土壤中漆酶活性的影响因素

针对纯化酶生产成本较高且土壤中漆酶活性较低的问题,通过20 d的室内对照实验,采用ABTS分光光度法测定漆酶活性,研究Cu2+、pH、温度、碳氮比及含水率对漆酶活性的影响,结果表明5个环境因素的最优值分别是40 mg·kg-1、5.2、30 ℃、40∶1和20%。环境因素与漆酶活性的高低有关,其相关性从大到小依次为pH、温度、含水率、碳氮比和铜离子浓度。其中,pH的相关性最高,相关系数为0.956。在最优条件下,培养30 d后石油烃降解率从自然条件下的8.54%提高到18.79%,最高漆酶活性达到13.97 U·g-1。     

应用漆酶SUKALacc脱色处理纺织染料

印染废水中含有大量的剩余染料,对生态环境构成了巨大威胁.漆酶能够氧化多种纺织染料使其脱色,在印染废水处理领域具有潜在的应用性.为了开发基于酶技术的印染废水处理工艺,利用商业化的漆酶制剂SUKALaccTM对5种常见纺织染料的脱色能力进行了测试,结果显示,SUKALaccTM对于这些测试的染料均具有脱色效果,脱色效率为14.85%~85.97%;此外,还考察了酶制剂浓度、染料浓度、温度和pH等因素对脱色效率的影响,确定了最优的反应条件;利用斜生栅藻 (Scenedesmus obliquus) 和大型溞 (Daphnia magna) 分别对酶处理后的染料毒性进行了测试,结果表明,酶制剂处理后的染料毒性减弱.这些实验结果为漆酶制剂实际应用于印染废水处理奠定了坚实基础.     

灵芝漆酶对直接蓝86的催化脱色性能

利用灵芝菌Ganoderma lucidum U-281漆酶对直接蓝86进行酶促氧化脱色,并对其降解机理进行了探讨。结果表明,染料-漆酶共反应体系在20～50℃及pH小于5.0范围内,直接蓝86均可脱色50%以上;漆酶对直接蓝86具有宽泛的浓度适应性,对300 mg/L的该染料仍具有耐受性。最优脱色工艺参数为温度40℃、pH 5.0、染料初始浓度200 mg/L、漆酶用量1 U/mL。在优化条件下,直接蓝863 h的脱色率达到54.54%,48 h脱色率达到91.54%。紫外-可见吸收光谱分析表明,漆酶的酶促氧化导致染料的分子结构产生了变化,是造成直接蓝86脱色的主要发生机制。     

生物炭和碳纳米管固定化漆酶去除水体雌二醇

漆酶是一类具有高效的催化氧化能力的含铜多酚的氧化酶,而固定化漆酶技术在有机废水处理领域受到了越来越多的关注。通过比较吸附法和包埋法利用木炭(WB)、竹炭(BB)和碳纳米管(CNT)固定游离漆酶去除水体雌二醇(E2)。在25 °C 、pH=5条件下,固定化漆酶经120 h 贮存仍保留80%的活性。经过5次循环研究,固载的漆酶仍保留了27%~58%的剩余活性。结合扫描电子显微镜和傅里叶变换红外光谱,证实了漆酶成功地负载在碳基材料上。仅在 8 h内,50 mg·L⁻¹ E2通过吸附法制备的WB、BB、CNT固定化漆酶去除了42.92%、47.66%、43.92%,而包埋法制备的WB、BB、CNT固定化漆酶分别去除84.08%、87.20%、87.86%。结果表明,包埋法固定化漆酶对E2的去除效率更优,这是由于包埋固载在生物炭和CNT的漆酶更为稳定。此外,K⁺和Mn²⁺离子可能通过干扰漆酶的电子传递过程影响漆酶的活性,从而抑制E2的去除。本研究旨在为固定化漆酶在有机废水处理的实际应用提供指导。     

Ganoderma sp. SYBC L48漆酶酶学性质及其对酸性红1的脱色性能

对灵芝菌Ganoderma sp. SYBC L48漆酶进行了纯化和酶学性质分析,并利用该漆酶对偶氮染料酸性红1进行脱色处理;考察了脱色体系中各因素对脱色效率的影响;采用小麦种子和水稻种子对酶处理后的染料进行了毒性测试。结果表明,以ABTS为底物时,该酶的最适pH为2.5,最适温度为60 ℃,在pH 5~9和20~60 ℃具有良好的稳定性,Co2+、Cr3+和Fe3+离子对酶活性有较强的抑制作用。在染料浓度100 mg·L-1,酶浓度0.5 U·mL-1,介体HOBT浓度0.25 mmol·L-1,pH为4,50 ℃的条件下反应30 min后,该漆酶对酸性红1的脱色率可达90.3%;1 mmol·L-1的Cr3+、Cu2+、Al3+和Ni2+存在下,漆酶仍能催化酸性红1脱色;脱色后染料的植物毒性下降。上述结果表明该漆酶在纺织废水处理中具有一定的应用前景。     

Factors affecting mass transfer limited biodegradation in saturated porous media

Microbial degradation rates in the subsurface are not only limited by the physiological capacity of the organisms, but also by inefficient supply of nutrients to the microbes. Although mass transfer limitation of biodegradation in the subsurface has been postulated for years, experimental evidence is still scarce. In the column experiments described here, diffusive transport of 4-nitroanisole from the bulk solution to cells of Rhodococcus opacus strain AS2 immobilized on glass beads or sand appeared to be responsible for the slow transformation rates observed. Assuming steady state, we applied a coupled transformation/transport equation to these data (Best equation) and apparent bead-related mass transfer coefficients were found to increase in proportion to the surface area covered with bacteria. This implies that mass transfer coefficients for individual cells remained constant. In an idealized oligotrophic environment where cells are only loosely clustered and do not shield each other, we would therefore expect biodegradation rates to be independent from the longitudinal distribution of the total biomass along a given flow path. Moreover, apparent mass transfer coefficients increased with the grain size of the column fillings, but did not change upon varying the flow rate. With a limiting external transport step, overall transformation fluxes do not become saturated at concentrations as low as predicted for Michaelis-Menten-type kinetics. Mass transfer limitation thus offers a justification for the common assumption that biodegradation rates in the subsurface follow first order kinetics in a wide concentration range.     

Simultaneous biodegradation of bifenthrin and chlorpyrifos by Pseudomonas sp. CB2

The degradation of bifenthrin (BF) and chlorpyrifos (CP), either together or individually, by a bacterial strain (CB2) isolated from activated sludge was investigated. Strain CB2 was identified as belonging to genus Pseudomonas based on the morphological, physiological, and biochemical characteristics and a homological analysis of the 16S rDNA sequence. Strain CB2 has the potential to degrade BF and CP, either individually or in a mixture. The optimum conditions for mixture degradation were as follows: OD_600nm = 0.5; incubation temperature = 30°C; pH = 7.0; BF-CP mixture (10 mg L^?1 of each). Under these optimal conditions, the degradation rate constants (and half-lives) were 0.4308 d^?1 (1.61 d) and 0.3377 d^?1 (2.05 d) for individual BF and CP samples, respectively, and 0.3463 d^?1 (2.00 d) and 0.2931 d^?1 (2.36 d) for the BF-CP mixture. Major metabolites of BF and CP were 2-methyl-3-biphenylyl methanol and 3,5,6-trichloro-2-pyridinol, respectively. No metabolite bioaccumulation was observed. The ability of CB2 to efficiently degrade BF and CP, particularly in a mixture, may be useful in bioremediation efforts.     

Decolorization and detoxification of two textile industry effluents by the laccase/1-hydroxybenzotriazole system

The aim of this work was to determine the optimal conditions for the decolorization and the detoxification of two effluents from a textile industry—effluent A (the reactive dye bath Bezactive) and effluent B (the direct dye bath Tubantin)—using a laccase mediator system. Response surface methodology (RSM) was applied to optimize textile effluents decolorization. A Box–Behnken design using RSM with the four variables pH, effluent concentration, 1-hydroxybenzotriazole (HBT) concentration, and enzyme (laccase) concentration was used to determine correlations between the effects of these variables on the decolorization of the two effluents. The optimum conditions for pH and concentrations of HBT, effluent and laccase were 5, 1 mM, 50 % and 0.6 U/ml, respectively, for maximum decolorization of effluent A (68 %). For effluent B, optima were 4, 1 mM, 75 %, and 0.6 U/ml, respectively, for maximum decolorization of approximately 88 %. Both effluents were treated at 30 °C for 20 h. A quadratic model was obtained for each decolorization through this design. The experimental and predicted values were in good agreement and both models were highly significant. In addition, the toxicity of the two effluents was determined before and after laccase treatment using Saccharomyces cerevisiae, Bacillus cereus, and germination of tomato seeds.     

Factors affecting ammonia volatilisation from a rice-wheat rotation system

Some of the major factors influencing ammonia volatilisation in a rice wheat rotation system were studied. A continuous airflow enclosure method was used to measure NH3 volatilisation in a field experiment at an agricultural college in Jiangsu Province. The five treatments comprised application rates of 0, 100, 200 or 300 kg N ha(-1) as urea, per growing season with rice straw amendment when wheat was sown, and 200 kg N ha(-1) without rice straw amendment. There were three replicates in a randomised block design. Ammonia volatilisation was measured immediately after urea application in the three consecutive years 1995 to 1997. The results show that N losses through NH3 volatilisation accounted for 4-19% of N applied during the wheat growing season and for 5-11% during the rice growing season. Ammonia volatilisation was affected significantly by soil moisture and temperature before and after fertiliser application during the wheat growing season. The ratio of volatilised NH3-N to applied N after urea application during the rice growing season was as follows: top-dressing at the onset of tillering > top-dressing at the start of the booting stage > basal fertilization. The results also show that the amount of N lost through NH3 volatilisation increased with increasing N application rate, but the ratio to applied N was not affected significantly by N application rate. Amendment with rice straw had no significant effect on NH3 volatilisation.     

开放静置体系中白地霉Y1对染料脱色降解研究

采用白地霉Y1对染料进行脱色,考察了染料初始浓度、接种量、pH值、C/N比和温度等因素对白地霉降解溴酚蓝的影响,并对其降解机理进行了探讨。实验结果表明：白地霉能在开放静置条件下对部分染料进行有效的脱色,对溴酚蓝尤其有效。开放静置体系下,白地霉能对300 mg/L的溴酚蓝进行脱色,脱色pH和温度分别为2～8、25～40℃,高碳低氮环境中更有利于脱色,接种量对染料脱色影响不大。通过对胞外液絮凝及脱色作用、葡萄糖代谢情况及染料脱色扫描图谱分析表明,白地霉主要通过分泌胞外液对溴酚蓝进行作用,且能对溴酚蓝有效降解。     

污泥酸化速率影响因子的探讨

采用城市污水厂污泥作为嗜酸微生物菌株来源,通过添加一定量的单质硫,使其中的嗜酸硫杆菌群大量增殖,并使污泥pH大幅降低.取得的培养物可用于废旧干电池中重金属沥滤等的处理.由于培养物对重金属沥滤效率和污泥的酸化速率密切相关,为此进行了不同的污泥种类、加硫量、污泥浓度和曝气强度对污泥酸化速率影响的实验.实验表明,初沉泥、二沉泥和混合浓缩污泥都能迅速利用硫产酸;加硫量(以100 mL污泥计)在0.5、1.0、2.0、4.0 g时,快速酸化的趋势相同.0.5 g的加硫量显示出略微慢的酸化速率,最后达到的最低pH在1.2左右,其他三个都降至1.0以下.污泥质量分数在0.5%、1.0%、2.0%、3.0%、4.0%时,也具有相同的酸化趋势,0.5%质量分数的污泥下降速率最快,这与较低浓度下污泥对pH的缓冲能力较小有关;曝气强度在0.45 L/min和0.3 L/min差别较小,5 d内能迅速降低pH,0.2 L/min的酸化速率较慢,足够长的时间(12 d)也能将pH降至2.5左右,0.1 L/min的曝气强度的酸化速率最慢,不能达到预期的酸化效果.     

Factors affecting xylene-contaminated air removal by the ornamental plant Zamioculcas zamiifolia

Fifteen plant species—Alternanthera bettzickiana, Drimiopsis botryoides, Aloe vera, Chlorophytum comosum, Aglaonema commutatum, Cordyline fruticosa, Philodendron martianum, Sansevieria hyacinthoides, Aglaonema rotundum, Fittonia albivenis, Muehlenbeckia platyclada, Tradescantia spathacea, Guzmania lingulata, Zamioculcas zamiifolia, and Cyperus alternifolius—were evaluated for the removal efficiency of xylene from contaminated air. Among the test plants, Z. zamiifolia showed the highest xylene removal efficiency. Xylene was toxic to Z. zamiifolia with an LC₅₀ of 3,464 ppm. Higher concentrations of xylene exhibited damage symptoms, including leaf tips turning yellow, holonecrosis, and hydrosis. TEM images showed that a low concentration of xylene vapors caused minor changes in the chloroplast, while a high concentration caused swollen chloroplasts and damage. The effect of photosynthetic types on xylene removal efficiency suggests that a mixture of Z. zamiifolia, S. hyacinthoides, and A. commutatum which represent facultative CAM, CAM, and C₃ plants, is the most suitable system for xylene removal. Therefore, for maximum improvement in removing xylene volatile compounds under various conditions, multiple species are needed. The effect of a plant’s total leaf area on xylene removal indicates that at lower concentrations of xylene, a small leaf area might be as efficient as a large leaf area.     

Physical factors negatively affecting evaluation of long-term biodegradation experiments of polychlorinated biphenyls

Behaviour of PCB added in the form of a technical mixture (Delor 106) and [¹⁴C]PCB 77 into different aqueous model cultivation media in long-term experiments was studied. The work was focused on monitoring PCB recovery from liquid media in the absence of micro-organisms. GC-ECD and radioanalysis were utilised for determination of PCB in the examined systems. Strong relations between duration of experiments, composition of media, degree of chlorination of biphenyls, and recovery of PCB were found. The lowest yields of PCB were observed for the extracts from model solutions based on N-limited cultivation media without veratryl alcohol and Tween 80. The addition of these components, mainly of Tween 80, caused a significant increase of PCB recoveries.     

脂肽生物表面活性剂作用下多菌种石油生物降解影响因素优化

探索多种菌种降解石油过程中菌种和脂肽生物表面活性剂的作用,筛选石油降解的主要影响因素及最佳组合,并为石油污染物的降解机理研究和石油污染修复提供指导。基于正交实验筛选主要影响因素,采用Box-Behnken实验探讨各因素最佳水平。正交实验中脂肽生物表面活性剂是多菌种降解石油过程中最主要的影响因素,在Box-Behnken实验中,其能显著地影响石油降解率。菌种降解能力是石油饱和烃组分生物降解的最主要影响因素,但脂肽生物表面活性剂是芳烃、胶质和沥青质组分降解的最主要的影响因素。研究所用菌种中,解淀粉芽孢杆菌（Bacillus amyloliquefaciens）和假单胞菌（Pseudomonas aeruginosa）在石油降解过程最重要,是本实验的石油降解最优菌。菌种和脂肽生物表面活性剂的添加浓度配比对于石油降解具有重要的影响。解淀粉芽孢杆菌和假单胞菌添加量5%,脂肽生物表面活性剂粗品添加量200 mg·L-1的降解效果最优,理论上,最高降解率可达63.78%,验证降解率达到了53.89%,相对于多菌种正交实验最高降解率提高了5.54%。利用正交实验和Box-Behnken实验筛选最优降解菌和最优菌种组合的方法,具有分析因素多、实验量少等优点,具有较好的应用前景。