产二羰基醛酮还原酶的1株嗜麦芽寡养单胞菌研究

用含甲基乙二醛的LB培养基平板筛选到1株能代谢甲基乙二醛的细菌,通过对克隆到的16S rDNA基因序列比对及生理生化实验,将其鉴定为嗜麦芽寡养单胞菌,命名为Stenotrophomonas maltophilia YL.破碎该菌细胞后提取粗酶,以甲基乙二醛作反应底物,在还原性辅酶Ⅰ存在条件下,紫外光谱和HPLC的分析表明,该菌能代谢甲基乙二醛;并对其产酶条件进行了研究.紫外分光光度法测得酶活力表明,该菌有较强的醛酮还原酶活力;并对温度和pH值对酶活力的影响作了分析.图5表1参17     

黑胸散白蚁纤维素酶的体外酶学特性

采用DNS法研究了我国广泛分布的一种低等木食性白蚁——黑胸散白蚁纤维素酶的体外酶活特性以了解其纤维素降解机制.结果表明,内切β-1,4-葡聚糖酶(EG)、纤维二糖水解酶(CBH)和β-葡萄糖苷酶(BG)这3种酶的最佳反应时间均为15 min,最佳底物浓度为1%,最适反应pH为5.6,最适反应温度为35℃.在最适反应条件下,EG、CBH和BG的活性分别达到71.3(±13.9)U/mg、5.8(±0.8)U/mg和4.1(±0.7)U/mg.EG在体外的热稳定性较差,在50℃及更高温度酶活很低或完全失活,但该酶对pH稳定性较好,在pH 3.2～8.0范围内酶活力变化不大.Native-PAGE电泳检测到该白蚁体内至少有8种不同的EG活性条带,肠道不同部位纤维素酶活性条带种类不同.这些研究表明,木食性白蚁降解纤维素是一个复杂的过程,需要多种纤维素酶的共同作用.     

一株产耐热纤维素酶菌株的筛选及酶学性质

采用纤维素刚果红平板法从大田蘑菇种植场建堆的稻草样中分离得到了1株能产具有较好温度耐受性和pH稳定性的纤维素酶的放线菌DY3.综合形态、生理生化特征以及16S rDNA序列分析,将其初步鉴定为嗜热裂孢菌(Thermobifida fusca).对该菌所产纤维素酶的性质研究表明:最适催化温度为65℃,在70℃保温60 min后仍有75%以上的活力;最适催化pH为7.5,在pH 5.5～10.0之间该酶的稳定性较好,pH 10.0的条件下仍有80%的活力;最适作用底物是羧甲基纤维素钠.研究可为木质纤维素的生物预处理提供一定参考价值.     

冰川环境耐冷菌的冷适蛋白酶分离纯化及酶学性质

采用离交和凝胶层析对“中国冰川1号”耐冷菌BacilluscereusSYPA23所产的蛋白酶进行分离纯化,并进行酶学性质研究.与中温酶相比,该蛋白酶具有较高的低温催化活力,其最适催化温度为42℃,适宜pH为7.0～8.5,SDSPAGE测定的分子量(Mr)为34.2×103.金属离子Mn2 、Ca2 对该酶有激活作用,Cu2 、Hg2 、Pb2 、Co2 对其活性有一定抑制作用.该酶属金属蛋白酶,其活性受EDTA强烈抑制,不受PMSF抑制.该蛋白酶具有低温酶典型的热不稳定性,0℃下半衰期24h,但Ca2 和一些低分子醇类物质能提高其稳定性.动力学数据分析表明,该蛋白酶在低温下对底物亲和能力高,在较宽温度范围内(25～45℃)均保持着较高的催化效能.图7表3参17     

一种新的中温酸性α-淀粉酶的分离纯化及酶学性质

从Bacillus sp.中分离纯化了一种新的中温酸性α-淀粉酶,并对其酶学性质进行了研究.粗酶经硫酸铵沉淀、 DEAE-Sepharose Fast Flow阴离子层析、 Sephadex G-75凝胶过滤层析等步骤后获得电泳均一的α-淀粉酶,其分子量(Mr)约为56×103.该酶最适反应pH为5.0,在pH 5.0～11.0范围内稳定,具有较好的耐酸特性.该酶最适反应温度为40～50 ℃,在40～60 ℃范围内稳定.在60 ℃、 pH 4.5时,该酶能快速有效水解可溶性淀粉;在pH 5.0时该酶对多种生淀粉底物也具有良好的水解作用.因此,该酶有助于解决中温条件淀粉加工行业中,液化酶和糖化酶由于适用pH值的差异而无法联用的难题,是一种具有研究价值和应用前景的中温酸性α-淀粉酶.经LC-MASS-MASS分析得到了酶蛋白中两个肽段的氨基酸序列,通过比对发现,该酶与NCBI中已报道的α-淀粉酶序列具有一定的同源性,同时,在氨基酸水平上也存在着差异.     

枯草芽孢杆菌(Bacillus subtilis)过氧化氢酶的分离纯化及性质

对产自枯草芽孢杆菌Bacillus subtilis WSHDZ-01的过氧化氢酶,通过乙醇沉淀、DEAE阴离子交换层析、疏水层析3步纯化,最终获得电泳纯的目标酶(纯化6.8倍).该过氧化氢酶的亚基相对分子质量(M1)为63×103,在405 nm处显示特征吸收峰,推测含有血红素.计算获得酶的表观米氏常数为26.87 mmol L-1.该过氧化氢酶不受低亚硫酸钠的还原作用影响,但被氰化物、叠氮化物和3-氨基-1,2,4-三唑(单功能过氧化氢酶的专一抑制剂)强烈抑制.以邻苯二胺作为电子供体测定酶活时,该酶不显示过氧化物酶活性,因此本文将纯化的过氧化氢酶定性为单功能过氧化氢酶.此外,该酶具有热敏感的特点,且酶活在pH 5～10范围内不受pH影响,此后,活性随着pH的升高而升高,并在pH 11～12处有明显的酶活高峰,在pH 11、25℃放置60 min酶活基本不变,表明该酶在高碱条件下具有很高的活力和一定的稳定性.     

提高天冬氨酸β-脱羧酶在酸性环境中催化活力的分子改造

克隆了德阿昆哈假单胞菌(Pseudomonas dacunhae)来源的L-天冬氨酸β-脱羧酶基因(Asd),实现其在Escherichiacoli中的异源表达,但该酶在酸性环境中活性较低,不利于工业生产.为通过定点突变技术提高该酶在酸性环境中的活力,选择底物通道区域内的5个氨基酸残基作为突变位点,构建6个突变体,随后分析突变体的酶学特性.结果表明,相比于野生型Asd,大多数突变体的比酶活显著下降,只有突变体N34D比酶活(71.67 U/mg)比野生型(65.95 U/mg)略高;另外,突变体N34D在3.5 pH 5.5酸性环境中的酶活力与野生型Asd相比均得到了提高,其中pH5.0条件下突变体N34D相对酶活达到82%,而野生型Asd相对酶活只有50%左右;而在pH 5.5-8.5范围内突变体N34D的酶活力与野生型相当.最后将突变体N34D应用于催化合成L-丙氨酸,在最适催化条件(pH 5.5)下,突变体N34D催化合成L-丙氨酸的产量是野生型Asd的1.5倍.本研究通过对德阿昆哈假单胞菌来源的N34位点进行突变,成功提高了天冬氨酸β-脱羧酶在酸性环境中的酶活力,并提高了其合成L-丙氨酸的能力,这对酶法生产L-丙氨酸具有重要的指导意义.     

漆酶对水中氯酚类物质的去除

利用漆酶对2,5-二氯苯酚(2,5-DCP)、2,6-二氯苯酚(2,6-DCP)、2,4,5-三氯苯酚(2,4,5-TCP)、2,4,6-三氯苯酚(2,4,6-TCP)等4种物质进行去除研究,考察了温度、时间、pH及漆酶添加量对去除效果的影响,并基于最佳条件下的去除效果,探讨了底物结构对漆酶转化氯酚类物质的影响.结果表明,温度升高,去除率呈先升高后降低再升高的现象;pH增大,去除率有增加的趋势;漆酶添加量在一定范围内与去除率呈正相关,当添加量高于20%后,去除率降低;在较短时间(0.5 h)内,漆酶对4种氯酚类物质的去除均能达到较好的效果.最佳条件下,漆酶对2,5-DCP、2,6-DCP、2,4,5-TCP和2,4,6-TCP的去除率分别为86.2%、45.3%、98.7%、84.8%;氯原子取代位置与取代数目相比对去除效果具有更重要的影响.     

耐碱性木聚糖酶产生菌的筛选及发酵条件研究

利用刚果红透明圈法,从造纸厂碱性土壤中筛选到一株木聚糖酶生产菌株X24-14,经培养特性研究及16S rDNA序列分析,初步认为该菌属于纤维菌属(Cellulosimicrobium).经发酵条件优化,该菌在麸皮60 g/L,蛋白胨10 g/L,K2HPO4 7.0 g/L, pH 8.5,接种量为5%, 37 ℃, 200 r/min的条件下发酵培养108 h,可达到最大活力,为2 204 u/mL. 该酶最适反应温度为60 ℃;具有较宽的pH作用范围,在pH 4.2～9.4范围内能保持较高的酶活力,在pH 9.4条件下,仍具有80%的酶活力; pH稳定性较好,在4 ℃、pH 11.0的条件下处理24 h仍能保持75%的酶活力.图3表1参12     

日本沼虾(Macrobrachium nipponense)N-乙酰-β-D-氨基葡萄糖苷酶初步纯化及部分性质

以日本沼虾内脏为材料,通过硫酸铵沉淀分级分离、DEAE-32离子交换柱层析和SephadexG-100分子筛柱层析纯化,获得比活力为3000Umg-1、纯化倍数为8.88倍的N-乙酰-β-D-氨基葡萄糖苷酶制剂(NAGase).以对-硝基苯-N-乙酰-β-D-氨基葡萄糖为底物,研究酶催化底物水解的反应动力学,结果表明,酶的最适pH为6.0,最适温度为53℃.该酶在pH4.5~9.3区域较稳定,当pH>9.3很快失活;在50℃以下处理30min,酶活力保持稳定,高于50℃,酶稳定性较差,75℃酶完全失活.酶促反应动力学符合米氏双曲线方程,测得米氏常数Km为0.165mmolL-1,最大反应速度Vm为6.55μmolL-1min-1.酶催化pNP-β-D-GlcNAc反应的活化能为63.55kJmol-1.图5表1参16     

酸雨对生菜的光合、抗氧化系统和产量的影响研究

目前国内外有关酸雨对植物生长危害的影响研究多集中在森林植物和粮食作物,而酸雨对叶菜类蔬菜生长的相关研究较少。酸雨的频发可能会影响蔬菜的生长,进而降低蔬菜产量。为探究酸雨对叶菜类蔬菜生理和产量的影响,选取生菜(Lactuca sativa L.var.ramosa Hort.)作为试验材料,采用野外原位研究方法,探究模拟酸雨(pH梯度为3.0、4.0、5.0)和自然降水(CK)对生菜光合、抗氧化系统和产量的影响。结果表明,pH 4.0和5.0酸雨对生菜光合系统影响不显著,pH 3.0酸雨对生菜光合系统影响显著;在整个试验期内,pH 3.0酸雨显著降低生菜净光合作用8.4%—17.0%。pH 3.0、4.0和5.0酸雨均显著提高生菜超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,表明酸雨会激活生菜抗氧化系统,同时会对生菜叶片膜系统造成损伤;在第20天后,pH 3.0酸雨下生菜过氧化氢酶(CAT)活性显著下降13.8%%—35.2%。pH 5.0酸雨对生菜产量影响不显著;pH 4.0和3.0酸雨显著降低生菜地上部产量,分别降低14.6%和16.7%。pH 3.0酸雨使生菜地下部产量显著降低19.0%。综上可见,模拟酸雨对生菜的光合系统、抗氧化酶系统和产量均产生显著影响,且酸雨对生菜的影响阈值在pH 4—5之间,pH 4.0以下的酸雨明显降低生菜产量,但酸雨强度为pH 5.0时已存在生菜减产的风险,研究结果可为生菜栽培的相关农户以及决策部门提供参考。     

Amelioration of Streamwater Acidity by Catchment Liming-Response of Podzolic Soils to Pasture Improvement

The effects of agricultural improvement by cultivation, re-seeding, lime and fertiliser additions on the soil solution chemistry of acid podzolic soils have been investigated in a field experiment in a mid-Wales catchment. the treatment resulted in an increase in the pH, alkalinity and calcium content of soil waters from the surface, organic-rich horizon. in the year following treatment, soil waters in the mineral horizons became more acid and had higher aluminium concentrations than observed at the control site. This response is ascribed to soil water acidification resulting from increased nitrification rates following liming and cultivation. in subsequent years, soil water acidity in these lower horizons was neutralised by lime-rich water percolating from the surface. No changes in stream water acidity and calcium content were observed in the catchment containing the area of agriculturally improved soils. This suggests that the selective treatment of moorland catchments to improve agricultural productivity may not always be accompanied by amelioration of acid stream water.     

Effects of water quality on the coagulation performances of humic acids irradiated with UV light

The presence of humic acid in drinking water treatment has received significant attention in recent years because of its adverse effects on the removal of many pollutants in coagulation. In this paper, the effects of water quality including pH, turbidity, alkalinity, and hardness on the removal of humic acid were investigated in a UV light hybridized coagulation process. Our results suggested that UV light radiation could effectively improve the removal rate of humic acid in coagulation under both neutral and basic conditions, and the variations of the selected water quality parameters had little adverse effect on the function of UV light. After UV light radiation, the removal rate of the nitro-humic acid (NHA) increased from 20% to 60% in coagulation, and increased further to 75% and 85% for the raw waters with 10.0 NTU kaolin and 100 mg·L^-1 hardness, respectively. In addition to NHA, the removal rates of the humic acid extracted from peat coal (PHA) and the humic acid provided by Japan metals and chemicals company (JHA) in coagulation were also improved, both in the range of 80%–90% after undergoing UV light radiation. By changing the radiation location from prior to coagulation to the flocculation process, similar experimental results were obtained. The formation of positive charged sites after UV light radiation was considered to be the primary factor that led to an enhanced removal of the humic acid in coagulation.     

吸附模式对有机物光催化降解的影响2.H-酸在TiO2表面的光催化降解途径

水溶液中H-酸通过磺酸基团吸附在TiO2表面,UV照射TiO2所产生的自由基首先进攻吸附在TiO2表面的磺酸基团,从而进一步导致萘环开环.pH 2.5条件下,虽然饱和吸附量较大,但由于H-酸仅通过一个磺酸基团吸附在TiO2表面,过程中产生的硫酸根速率较慢,最终的光降解速率也较慢.pH 5.0条件下,虽然饱和吸附量较小,但由于吸附在TiO2表面的两个磺酸基团同时受到来自TiO2表面自由基的进攻,过程中产生的硫酸根速率较快,最终的光降解速率也较快.吸附模式的差异是导致H-酸在不同pH值条件下光催化降解途径和速率差异的关键因素.     

Acid resistance of methanogenic bacteria in a two-stage anaerobic process treating high concentration methanol wastewater

In this study, the two-stage upflow anaerobic sludge blanket (UASB) system and batch experiments were employed to evaluate the performance of anaerobic digestion for the treatment of high concentration methanol wastewater. The acid resistance of granular sludge and methanogenic bacteria and their metabolizing activity were investigated. The results show that the pH of the first UASB changed from 4.9 to 5.8 and 5.5 to 6.2 for the second reactor. Apparently, these were not the advisable pH levels that common methanogenic bacteria could accept. The methanogenic bacteria of the system, viz. Methanosarcina barkeri, had some acid resistance and could still degrade methanol at pH 5.0. If the methanogenic bacteria were trained further, their acid resistance would be improved somewhat. Granular sludge of the system could protect the methanogenic bacteria within its body against the impact of the acidic environment and make them degrade methanol at pH 4.5. The performance of granular sludge was attributed to its structure, bacteria species, and the distribution of bacterium inside the granule.     

The Effects of Acidity on Carbon Fluxes from Ombrotrophic Peat

Replicate intact peat/vegetation monoliths were collected along a pollution gradient in the UK and subjected over one year to 1200 mm of simulated rainfall of the same chemical composition as they received in the field. Drainage water was analysed periodically for DOC and pH, and the decomposition rates of Calluna vulgaris and Eriophorum angustifolium leaves were measured, as well as soil atmosphere CO₂ contents and peat matric potentials. the chemical characteristics of the peats as initially sampled from the field also were determined. the results suggest that acidic precipitation has induced chemical changes in ombrotrophic peats, lowering their pH and base status, when due account is taken of calcium deposition or any mineral content. Greater DOC fluxes were observed from the more acid peats, and litter decomposition rates from these peats were reduced.     

Decomposition of perfluorooctanoic acid by microwaveactivated persulfate: Effects of temperature, pH, and chloride ions

Microwave-hydrothermal treatment of persistent and bioaccumulative perfluorooctanoic acid (PFOA) in water with persulfate (S₂O ₈ ^2? ) has been found effective. However, applications of this process to effectively remediate PFOA pollution require a better understanding on free-radical scavenging reactions that also take place. The objectives of this study were to investigate the effects of pH (pH = 2.5, 6.6, 8.8, and 10.5), chloride concentrations (0.01?C0.15 mol·L^?1), and temperature (60°C, 90°C, and 130°C) on persulfate oxidation of PFOA under microwave irradiation. Maximum PFOA degradation occurred at pH 2.5, while little or no degradation at pH 10.5. Lowering system pH resulted in an increase in PFOA degradation rate. Both high pH and chloride concentrations would result in more scavenging of sulfate free radicals and slow down PFOA degradation. When chloride concentrations were less than 0.04 mol·L^?1 at 90°C and 0.06 mol·L^?1 at 60°C, presence of chloride ions had insignificant impacts on PFOA degradation. However, beyond these concentration levels, PFOA degradation rates reduced significantly with an increase in chloride concentrations, especially under the higher temperature.     

模拟酸雨对酸性土壤铝溶出及其形态转化的影响

研究了我国南方酸沉降区主要土壤类型在模拟酸雨影响下,土壤中铝离子释放及铝形态转化的特点。结果表明,酸雨淋洗造成土壤中铝离子释放,酸雨pH值越低,铝离子释放量越大;酸雨淋洗还造成土壤中铝形态发生变化,酸雨pH越低,土壤中羟基态铝和腐殖质铝含量愈低,交换态铝含量越高,土壤中铝对植物和生态系统的危害性也越大。     

模拟酸雨对华南典型树种生长及营养元素含量的影响

我国是世界上的第3大酸雨区,而华南地区是我国酸雨比较严重的地区之一。越来越严重的酸雨将对森林生态系统林木生长产生负面影响。本文采用盆栽试验模拟研究酸雨对华南典型树种尾叶桉和马尾松幼树生长及营养元素含量的影响。酸雨水平有4个:pH6.8(CK)、pH5、pH4和pH3。幼树生长期间观察并记录其酸雨受害症状,每隔10a测量一次树高,4个月后统计生物量、叶面积情况,并分析植株样品中的N、P、K、Ca、Mg等元素的质量分数。试验结果表明,在不同酸度酸雨的协迫下,尾叶桉和马尾松幼苗表现出不同程度的受害症状,酸度越高受害越严重。酸雨显著地抑制尾叶桉和马尾松的树高生长,降低幼苗生物量,减少叶面积生长。与对照相比,生长四个月后,pH5、pH4和pH3处理分别导致尾叶桉生物量降低10.79%、19.21%和27.25%,马尾松降低12.78%、22.98%和28.70%。尾叶桉植株内P、Mg、Ca质量分数不同程度地受到酸雨的影响,而N、K质量分数不受酸雨影响;马尾松除了P元素以外,其它元素质量分数在各个酸雨处理之间差异不显著。     

利用非活体生物质去除废水中重金属的研究

为了解非活体牛物质去除废水中重金属的效果，选择了玉米芯、水稻谷壳、花生壳、松树树皮和茶叶等5种非活体生物质，州室内模拟方法比较研究了它们对重金属的吸附能力。结果表明，生物质对重金属的吸附是一个快速反应，可在20-30min内达到平衡。pH值对牛物质吸附阳离子型重金属有很大的影响，吸附量随pH值上升而增加，pH值在4．5以上时可达到较高水平。5种生物质对重金属都有较高吸附能力，它们吸附重金属的能力依次为：花生壳〉松树树皮〉玉米芯〉水稻谷壳〉茶叶。生物质去除废水中重金属的效果一般为：Cu、Pb〉Cd〉Zn。用碱、柠檬酸和磷酸对生物质进行改性处理可显著增强其对重金属的去除能力。生物质是一种廉价、有效的吸附剂，可替代商品吸附剂用于废水中重金属的去除，主要重金属的移除率在85％以上。