Myrothecium verrucaria NF-08产漆酶条件的响应面法优化及粗酶染料脱色研究

以半知菌属疣孢漆斑菌Myrothecium verrucaria NF-08为供试菌,采用响应面方法优化菌株产漆酶条件,并以其漆酶粗酶进行3种结构类型6种染料的脱色试验,考察了粗酶浓度、温度和p H值对脱色率的影响.两水平析因设计、中心组合设计和验证试验结果表明:葡萄糖浓度为15.4 g·L~(-1)、蛋白胨浓度为43.3 g·L~(-1)和培养基初始p H值为6.68时,漆酶活性达到最大值(25.58±1.60)U·m L~(-1),是单因素试验(16.82 U·m L~(-1))的1.52倍,提高了52.1%.以上述发酵条件获得的M.verrucaria NF-08漆酶粗酶对3种结构类型染料均有脱色效果,排序为偶氮类优于芳甲烷类优于蒽醌类.脱色率随粗酶浓度增大(2~8 U·m L~(-1))、反应温度升高(15~35℃)而上升,随p H值上升(4~10)而下降.粗酶浓度为5 U·m L~(-1),反应温度为25℃,p H值为4时,偶氮类染料铬黑T和芳甲烷类染料碱性品红即可几乎完全脱色,脱色率分别为99.2%和94.6%.     

基因工程菌漆酶对蒽醌染料的脱色研究

利用基因工程菌甲醇毕赤酵母(PMAD16/pMETαA-Lcc1)培养产生的重组漆酶,研究了重组漆酶对蒽醌染料Remazol Brilliant Blue R脱色的影响。结果表明:重组漆酶对蒽醌染料RBBR脱色的最佳pH值为4.5,温度为45℃,当溶液中漆酶活力为10.0U/mL时,在最适脱色条件下作用14h,粗漆酶和纯化漆酶对蒽醌染料RBBR(80mg/L)的脱色率分别为95.2%和87.5%,重组漆酶可有效地使蒽醌染料RBBR脱色。     

Trametes sp.LS-10C产漆酶发酵培养基优化及其漆酶对偶氮染料的脱色性能

为了提高菌株Trametes sp.LS-10C的漆酶产量并初步研究该酶对偶氮染料的脱色性能,本文通过单因素实验及均匀设计对菌株Trametes sp.LS-10C产漆酶培养基进行了优化,获得最优培养基配方为:葡萄糖14.4 g·L~(-1)、豆渣10.1 g·L~(-1)、麸皮100 g·L~(-1)、NH_4Cl_3g·L~(-1)、KH2PO41.4g·L~(-1)、CuSO_4’5H2O 1 g·L~(-1)、NaCl 1 g·L~(-1)、MgSO_40.8 g·L~(-1)、CaCl_20.5 g·L~(-1).菌株发酵至8 d时漆酶产量为595.15 U·m L~(-1),约是优化前的35.66倍.进一步研究表明,该漆酶-介体系统对酸性红GR、酸性蓝40和酸性铬蓝K等3种偶氮染料脱色10 h后脱色率分别高达96.86%、91.28%和86.31%.相比公开发表的文献,本研究所报道的Trametes sp.LS-10C利用优化培养基发酵产漆酶达到了较高水平,具有酶活力高和发酵时间短等特征,且该漆酶在处理偶氮染料废水脱色领域中具有进一步研发和应用价值.     

大孔树脂共价固定化漆酶及其对孔雀石绿的脱色研究

以大孔树脂为固定化载体,戊二醛为表面活化剂,采用共价结合法固定漆酶。以ABTS为漆酶底物,通过对固定化漆酶活性的测定,确定大孔树脂固定漆酶的最佳固定条件为：戊二醛浓度=2%、pH=6、活化时间=2 h、固定时间=10 h。在此条件下制备的固定化漆酶能够保持79%的初始活性,获得的固定化漆酶活力能够达到1．94 U·g-1。在酸性条件(pH=3)下,游离漆酶的活性在3 h内丧失71%,而固定化漆酶仅丧失37%,显示出固定化漆酶良好的环境稳定性。固定化漆酶对孔雀石绿染料的脱色率在5h内可达到96%。     

蜜环菌漆酶对蒽醌类染料的脱色条件优化

利用蜜环菌发酵所得的漆酶,直接对印染工业中常见的两种蒽醌类染料活性艳蓝KN-R和活性艳蓝X-BR进行催化脱色实验,得出了最佳脱色条件.结果表明,活性艳蓝KN-R最适脱色温度为30℃,最适染料浓度为80 mg.L-1,最适酶量为0.25U.mL-1,最适pH值为5,在最优条件下活性艳蓝KN-R最高脱色率达90%以上.活性艳蓝X-BR的最适脱色温度为30℃,最适染料浓度为50 mg.L-1,最适酶量为0.5 U.mL-1,最适pH值为4,在最优条件下活性艳蓝X-BR最高脱色率达70%以上.本研究利用蜜环菌粗漆酶液直接对印染工业中常见蒽醌类染料进行脱色,结果表明蜜环菌粗漆酶液具有良好的脱色效果,蜜环菌漆酶在印染工业染料废水脱色方面具有潜在的应用价值.     

红芝所产漆酶对蒽醌染料的脱色研究

以红芝Ganoderma lucidum发酵所得漆酶粗酶液对蒽醌染料进行脱色实验,分别考察了反应时间、染料浓度、温度、pH、转速等条件对蒽醌染料脱色的影响。结果表明,红芝漆酶粗酶液对蒽醌染料具有很好的脱色效果,脱色的适宜条件为:染料浓度150mg/L、反应温度40℃、pH值3.5。在上述条件下反应8h,蒽醌染料脱色率可达90%左右,摇床转速对染料脱色影响不大。     

黄曲霉A5p1脱色类黑精及其机理初探:葡萄糖氧化酶的作用

类黑精是一种高分子难降解色素污染物质,在糖蜜酒精废水中大量存在.选取在前期研究中对糖蜜酒精废水具有较好脱色作用的黄曲霉(Aspergillus flavus)A5p1(保藏号CGMCC.4292),以合成的类黑精为对象研究脱色机理,试图为实际废水的生物脱色提供理论基础.结果显示,此菌株对合成类黑精具有生物吸附和生物降解的双重作用,以后者为主,最高脱色率可达65%.相较于漆酶、依赖/不依赖于锰的过氧化物酶等常规氧化脱色酶,此菌株中的产过氧化氢酶对脱色起主要作用,而且脱色过程中作为产过氧化氢酶之一的葡萄糖氧化酶与类黑精脱色率之间存在着一定的正相关性.添加葡萄糖氧化酶激活剂和抑制剂进行验证,发现葡萄糖氧化酶酶活力、过氧化氢产量和脱色率三者之间呈相关性变化.初步推测,葡萄糖氧化酶是黄曲霉A5p1生物降解脱色类黑精的关键酶之一.     

漆酶介体系统对孔雀绿的脱色研究

研究了漆酶介体系统对代表性三苯甲烷类染料孔雀绿的脱色处理。漆酶介体系统对孔雀绿的脱色效率比单独使用漆酶要高出10倍。高浓度的孔雀绿对漆酶介体系统的脱色效率有抑制作用,最适的pH值和温度分别在4.2和32°C左右。介体浓度的提高可促进脱色率的增加,但高浓度的介体会抑制脱色效率。漆酶和漆酶介体系统对孔雀绿的脱色机制上存在不同,前者的作用机制为脱甲基反应,而后者的脱色机制为自由基介导的聚合反应。     

Aureobasidium pullulans UVMU3-1产耐热性胞外漆酶活性物质培养基优化及其对染料脱色作用

对出芽短梗霉无色素突变株Aureobasidium pullulans UVMU3-1产耐热性胞外漆酶活性物质培养基进行优化,并研究其分子量大小、热稳定性及对三苯甲烷类染料孔雀石绿和结晶紫的脱色作用.结果表明:蔗糖、NaNO_3为UVMU3-1产耐热性胞外漆酶活性物质的最佳碳源和氮源,产酶优化培养基配方为:蔗糖66.5 g·L~(-1),NaNO_338.4 g·L~(-1),NaCl 1.08 g·L~(-1),酵母浸粉0.2 g·L~(-1),KH_2PO_41 g·L~(-1),pH=6.0.影响UVMU3-1产耐热性胞外漆酶活性物质的因素依次为:起始pH蔗糖NaClNaNO_3,培养7 d时产生最大酶活(94.81 U·L~(-1)),比对照提高3.85倍.其分子量小于1000 D,具有良好的热稳定性,煮沸30 min后高温酶活仍残留72.50%.粗酶液(7.5 U·L~(-1))与三苯甲烷类代表性染料孔雀石绿和结晶紫75℃反应5 min后,脱色率分别为76.3%、77.5%,反应35 min后脱色率分别达到96.3%、95.3%.     

氨基酸修饰的高交联吸附树脂对Cu(Ⅱ)和磺胺嘧啶的吸附机制研究

采用络氨酸修饰氯球树脂（PS-DVB）并进行后交联反应制备了氨基酸修饰的高交联吸附树脂（PSTY）.研究了PSTY树脂对Cu（Ⅱ）和磺胺嘧啶（SDZ）的协同吸附性能及交互影响机制.单组分体系中,树脂对Cu（Ⅱ）的吸附随pH值的增加而增加,对SDZ的吸附随着pH值的增加先增后减;共存体系中,SDZ的共存微弱促进了树脂对Cu（Ⅱ）吸附,Cu（Ⅱ）的共存抑制了树脂对SDZ的吸附,且pH大于6后抑制更为明显.PSTY对Cu（Ⅱ）和SDZ的吸附等温线可采用Langmuir和Freundlich方程进行拟合,共存低浓度的SDZ使Cu（Ⅱ）的饱和吸附量略有增加,而高浓度的SDZ使其下降.Cu（Ⅱ）的增强去除主要是阴离子SDZ^-与树脂的质子化基团的静电作用减弱了对Cu（Ⅱ）的静电斥力,Cu（Ⅱ）与SDZ分别与PSTY中的极性基团发生配位作用和氢键作用进行竞争吸附,但Cu（Ⅱ）的配位结合力更强,虽然吸附到树脂上的Cu（Ⅱ）还可以通过桥连作用吸附SDZ,但作用较弱,因此,共存的Cu（Ⅱ）抑制了PSTY树脂对SDZ的吸附,Cu（Ⅱ）浓度越大抑制程度越高.     

乙醇处理对树脂基纳米水合氧化铁结构及其除砷性能的影响

通过考察乙醇处理对树脂基纳米水合氧化铁(D201-HFO)结构及其除砷性能的影响,评价乙醇处理在复合材料制备中的必要性.结果表明,乙醇处理后负载水合氧化铁(HFO)在D201树脂内的分散性增加,材料对砷的饱和吸附量提高约20%,且复合材料的比表面积、孔容、孔径分别增加了52%、65%、28%;但乙醇处理对复合材料的机械强度和负载HFO的晶型无影响,在pH值、离子竞争对该材料除砷性能的影响趋势方面也无明显作用,对复合材料固定床除砷性能及再生方面也无明显作用.总体而言,乙醇处理对D201-HFO复合材料实际除砷性能影响不大,从技术经济性考虑可以省略.     

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.     

环氧化PVA载体固定漆酶的最佳工艺条件研究

在T=35℃,HAc-NaAc缓冲溶液pH为4的条件下,用环氧化PVA载体对漆酶进行固定化,固定后酶活为游离酶活的95%。用该固定化漆酶处理制浆造纸废水,反复使用6次后,酶活仍保持为初始酶活的45%,且每次COD去除率在40%以上,处理后的废水BOD增大了50%,废水可生化性得到很大提高。     

纳米二氧化钛对三价砷在大型蚤体内累积与毒性的影响

nTiO₂（纳米二氧化钛）可作为其吸附污染物的运输载体而影响污染物在生物体内的累积与毒性,为明晰nTiO₂吸附As（Ⅲ）（三价砷）后作为As的运输载体对水生物体内As累积与毒性的影响,以大型蚤（Daphnia magna）为受试生物,通过室内培养试验,研究了不同nTiO₂浓度、As（Ⅲ）不同暴露水平相互作用下蚤体中As与Ti的累积含量、毒性及其潜在作用机制.结果表明,nTiO₂可在30 min内吸附As（Ⅲ）至平衡,其中2、20 mg/L的nTiO₂对75 μg/L As（Ⅲ）的吸附率分别可达31.38%、51.84%,蚤体内As的累积含量分别为对照组的2.9和3.8倍,表明nTiO₂可作为运输载体提高As（Ⅲ）在大型蚤体内的累积;但不同As暴露水平下蚤体As和Ti含量的相关关系表明,nTiO₂作为载体的运输作用可能会因As暴露水平的增加而减弱.另外,nTiO₂虽然作为运输载体提高了As（Ⅲ）在大型蚤体内的累积,但并未增加As（Ⅲ）对大型蚤的毒性.添加2、20 mg/L的nTiO₂后,As（Ⅲ）对大型蚤的24 h IC₅₀（半抑制浓度）分别从0.93 mg/L增至2.53和2.97 mg/L,表明nTiO₂降低了As（Ⅲ）对大型蚤的毒性.研究显示,nTiO₂虽增加了As（Ⅲ）在大型蚤体内的累积,但却降低了As（Ⅲ）对大型蚤的毒性,这有利于对nTiO₂及其复合重金属污染风险的深入认识.      

Cd对不同形态漆酶修复DDT污染土壤的影响

研究了重金属Cd对游离漆酶和同定化反胶团漆酶修复土壤有机氯农药DDT污染的影响.结果表明.游离漆酶对土壤中DDT各组分均有不同程度降解,且均随着cd浓度的增大而降低,含量越高的组分降解率越高.受到Cd污染的影响也越大(P,P'-DDT>P,P'-DDD>0.P'-DDT>P,P'-DDE);当Cd浓度分别为0、0.5、1和2 mg·kg-1时,游离漆酶对土壤中DDT总量的降解率分别为50.68%、32.50%、14.92%和13.40%.固定化反胶团漆酶比游离漆酶能更有效地降解DDT,DDT降解率在无Cd和有Cd存在时(Cd浓度为0.5 mg·kg-1)分别提高20%和30%左右.     

固定化真菌对铅离子生物吸附的研究

采用多种固定化材料包埋枝孢霉属(Cladosporium sp.),结果表明:以一定比例混合的海藻酸钠-明胶包埋的枝孢霉小球对Pb2+具有最佳吸附效果。通过正交试验确定了固定化最优操作条件,最优条件为:菌量为10g/L,包埋剂量为1.5∶1.5,CaCl2浓度为4%,钙化时间为4h。并考察了不同因素如接触反应时间,溶液的pH,温度对生物吸附的影响。结果表明:平衡吸附的时间为3h左右,最适pH为4.0,在15℃~45℃的温度范围内,吸附量有缓慢增加,在一定的浓度范围(30~700mg/L)内,生物吸附随浓度的增加而增加,生物吸附过程较好地符合Langmuir模型。     

Biosorption characteristics of unicellular green alga Chlorella sorokiniana immobilized in loofa sponge for removal of Cr(Ⅲ)

Loofa sponge (LS) immobilized biomass of Chlorella sorokiniana (LSIBCS), isolated from industrial wastewater, was investigated as a new biosorbent for the removal of Cr(Ⅲ) from aqueous solution. A comparison of the biosorption of Cr(Ⅲ) by LSIBCS and free biomass of C. sorokiniana (FBCS) from 10-300 mg Cr(Ⅲ)/L aqueous solutions showed an increase in uptake of 17.79% when the microalgal biomass was immobilized onto loofa sponge. Maximum biosorption capacity for LSIBCS and FBCS was found to be 69.26 and 58.80 mg Cr(Ⅲ)/g biosorbent, respectively, whereas the amount of Cr(Ⅲ) ions adsorbed onto naked LS was 4.97 mg/g. The kinetics of Cr(Ⅲ) biosorption was extremely rapid and equilibrium was established in about 15 and 20 min by LSIBCS and FBCS,respectively. The biosorption equilibrium was well defined by Langmuir adsorption isotherm model. The biosorption kinetics followed the pseudo-second order kinetic model. The biosorption was found to be pH dependent and the maximum sorption occurred at the solution pH 4.0. Desorption studies showed that 98% of the adsorbed Cr(Ⅲ) could be desorbed with 0.1 mol/L HNO3, while other desorbing agents were less effective in the order: EDTA ＞ H2SO4 ＞ CH3COOH ＞ HCl. The regenerated LSIBCS retained 92.68% of the initial Cr(Ⅲ) binding capacity up to five cycles of reuse in continuous flow-fixed bed columns. The study revealed that LSIBCS could be used as an effective biosorbent for the removal of Cr(Ⅲ) from wastewater.     

Effect of ionic strength on adsorption of As（Ⅲ） and As（Ⅴ） on variable charge soils

The study was to investigate the adsorption behavior of arsenite (As(Ⅲ)) and arsenate (As(Ⅴ)) on two variable charge soils, i.e., Haplic Acrisol and Rhodic Ferralsol at different ionic strengths and pH with batch methods. Results indicated that the amount of As(Ⅲ) adsorbed by these two soils increased with increasing solution pH, whereas it decreased with increasing ionic strength under the acidic condition. This suggested that As(Ⅲ) was mainly adsorbed on soil positive charge sites through electrostatic attraction under the acidic condition. Moreover, intersects of As(Ⅴ) adsorption-pH curves at different ionic strengths (a characteristic pH) are obtained for both soils. It was noted that above this pH, the adsorption of As(Ⅴ) was increased with increasing ionic strength, whereas below it the reverse trend was true. Precisely the intersect pH was 3.6 for Haplic Acrisol and 4.5 for Rhodic Ferralsol, which was near the values of PZSE (soil point of zero salt effect) of these soils. The effects of ionic strength and pH on arsenate adsorption by these soils were interpreted by the adsorption model. The results of zeta potential suggested that the potential in adsorption plane becomes less negative with increasing ionic strength above soil PZSE and decreases with increasing ionic strength below soil PZSE. These results further supported the hypothesis of the adsorption model that the potential in the adsorption plane changes with ionic strength with an opposite trend to surface charge of the soils. Therefore, the change of the potential in the adsorption plane was mainly responsible for the change of arsenate adsorption induced by ionic strength on variable charge soils.     

壳聚糖-海藻酸钠固定化菌小球处理猪场沼液

采用包埋法,以海藻酸钠-壳聚糖为载体、自行筛选的高效脱氨氮菌为目标菌制备固定化菌小球,用于去除猪场沼液中的氨氮.优化了固定化小球的制备条件,考察了废水处理条件对氨氮去除效果的影响.结果表明,固定化菌小球的最佳制备条件为：壳聚糖投加量20g/L、海藻酸钠投加量10g/L、目标菌种包埋量2：5（V/V）.处理含氨氮废水时,在不调节废水pH值的条件下,当固定化菌小球投加量为15g/L、反应时间为4h时,氨氮的去除率为93.9%,其中吸附作用对氨氮的去除率为64.3%,微生物作用对氨氮的去除率为29.6%.扫描电镜表征结果表明,处理废水后,固定化菌小球外部及内部微生物数量明显增多.动力学与等温线拟合结果显示,固定化菌小球对废水中氨氮的去除过程符合准二级反应动力学方程（R²=0.9252）和Langmuir等温线方程（R²=0.9578）.     

海藻酸钠凝胶球去除单宁酸和没食子酸的研究

采用海藻酸钠-钙凝胶球、海藻酸钠-钙-铁(Ⅲ)凝胶球、海藻酸钠-铁(Ⅲ)凝胶球对水体中单宁酸和没食子酸进行吸附实验研究。结果表明,对50mL(50mg/L)的模拟污水进行吸附,海藻酸钠-铁凝胶球和海藻酸钠-钙-铁(Ⅲ)凝胶球的吸附效果优于海藻酸钠-钙凝胶球,吸附120min后,使用Fe(Ⅲ)凝胶球对单宁酸的去除率可达97%,对没食子酸的去除率可达99%,对两种物质的吸附量分别为8.0、8.3mg/g,使用海藻酸钠-钙-铁(Ⅲ)凝胶球对单宁酸、没食子酸的去除率可达84%和97%,对两种物质的吸附量分别为7.0、8.0 mg/g。饱和吸附后海藻酸钠-钙-铁(Ⅲ)凝胶球可以再生利用,对单宁酸的去除效果随着降解再生次数的增加而降低。