Cd胁迫下超富集植物圆锥南芥抗氧化机理

采用土培的方法研究了不同Cd2+添加量(0、10、20、40、80、160、240mg.kg-1)对多金属超富集植物圆锥南芥(Arabis paniculata Franch)抗氧化酶(超氧化物歧化酶SOD、过氧化氢酶POD、抗坏血酸过氧化物酶CAT、过氧化物酶APX)活性、非酶物质(巯基SH、谷胱甘肽GSH、植物络合素PCs)含量以及生物量、叶绿素、丙二醛(MDA)、半胱氨酸(Cysteine)、脯氨酸(Proline)含量的影响.结果表明,随着Cd添加量的增加,圆锥南芥的生物量、Cd含量、富集系数、转运系数呈增加趋势,SOD活性、叶绿素含量呈下降趋势.POD、CAT、APX活性以及MDA含量均呈先升高后降低的变化趋势.圆锥南芥叶片中SH、GSH、PCs含量随Cd添加量的增加而增加,当Cd添加量为240mg.kg-1时,SH、GSH、PCs含量分别比对照提高了92.7%、70.2%、143.4%,表明SH、GSH、PCs在解毒Cd毒害中起着重要作用.     

Expression of lignin peroxidase H2 from Phanerochaete chrysosporium by multi-copy recombinant Pichia strain

The lipH2 gene,encoding the expression of lignin peroxidase,was cloned from Phanerochaete chrysosporium BKM-F-1767 and expressed in Pichia pastoris X-33,a yeast.The cDNA of LiPH2 was generated from total RNA extracted from P.chrysosporium by PCR with primers that do not contain a P.chrysosporium lignin peroxidase secretion signal.The gene was then successfully inserted into the expression vector pPICZα,and resulted in the recombinant vector pPICZα-lipH2.The transformation was conducted in two ways.One was u...     

Interactive e ect of dissolved organic matter and phenanthrene on soil enzymatic activities

The investigation of dissolved organic matter (DOM) on urease, catalase and polyphenol oxidase activity in a phenanthrene (Phe)- contaminated soil was conducted under laboratory incubation conditions. Values of soil enzymatic activity depended mainly on incubation time. In the initial 16 days, urease activity increased, and was followed by a decrease. In the initial 8 days, catalase activity decreased and then increased. Variation of polyphenol oxidase activity was just the reverse of catalase activity. After 30 days of incubation, no pronounced di erence among treatments with Phe, Phe and DOM, and control were detected in urease, catalase and polyphenol oxidase activity. Phe might inhibit urease and catalase, and stimulate polyphenol oxidase. DOM could improve inhibition of Phe in soil urease and catalase activity during the initial period of applying DOM. Nevertheless, DOM had no significant e ect on polyphenol oxidase activity in the Phe contaminated soil. There was a negative correlation between catalase and polyphenol oxidase (r = –0.761***), and catalase and urease (r = –0.554**). Additionally, a positive correlation between polyphenol oxidase and urease was also detected (r = 0.701***). It is implied that the formed DOM after application of organic wastes into soils may counteract the inhibition of polycyclic aromatic hydrocarbons in soil enzyme activities.     

三苯基甲烷类染料氧化酶基因(tpmD)在大肠杆菌中的无诱导表达

为了解决嗜水气单胞菌DN322对鱼类的潜在致病性问题,有必要克隆表达该菌的三苯基甲烷染料脱色酶基因tpmD.通过PCR方法获得该基因,并与脱色希瓦氏菌S12的NAD(P)H脱氢酶基因启动子和用于荧光标记的编码CCPGCC的碱基序列融合,将有启动子和无启动子的融合基因片段分别连接到质粒pMD18-T中,转化大肠杆菌E.coli DH5α和E.coli BL21.结果发现,有启动子的融合基因能被E.coil在不加诱导物IPTG的条件下高效正确表达,脱色酶活性甚至超过基因供体菌DN322.将上述两株大肠杆菌工程菌株在自然水体中进行孔雀石绿污染小试处理,60d内在每个350mL反应体系中累积共投加大于10000mg的孔雀石绿,脱色率一直保持在92%以上;细菌数量最后增加了5～10倍.研究结果证明,基因工程菌在不加营养物的条件下也有很强的脱色活性和长期的存活能力;在tpmD基因3'端加入的用于螯合双砷荧光染料的18bp编码氨基酸碱基序列不影响此基因的表达和酶的脱色活性,这将赋予此工程菌可示踪性.     

H2O2存在条件下冰相中苯酚的光化学转化

研究了在125W高压汞灯照射下,低温(-14～-12℃)反应装置中H2O2存在条件下冰相中苯酚的光转化反应,考察r冰相中苯酚光转化的影响因素以及光转化动力学过程.结果表明,冰相中苯酚光转化速率随着光强和H2O2初始浓度的增加而加快,随着苯酚初始浓度的增加而减慢;强酸条件促进了苯酚的光转化;苯酚的光解速率符合一级动力学模式.苯酚光解产物的GC-MS和LC-MS分析表明,苯酚在光解过程巾主要发乍了羟基化反应和偶联反应.     

四溴双酚-A污染对颤蚓的氧化胁迫及毒性

在实验室模拟条件下分别采用动、静态生物慢性实验的方法,研究了不同浓度的四溴双酚-A(Tetrabromobisphenol A, TBBPA)对颤蚓(Monopylephorus limosus)体内抗氧化酶系统及相Ⅱ结合酶系统活性的影响以及颤蚓暴露于单一浓度的TBBPA后,其体内抗氧化酶系统的活性随时间的响应.结果表明,暴露 8d 后,颤蚓体内SOD的活性发生显著变化,变化趋势明显分为升高、降低、再升高3阶段,在0.05ms/L时SOD的活性被最大诱导(p<0.01),达到对照组的7.8倍,各浓度组SOD的活性均显著高于对照组,由对照组的1.5倍到对照组的7.8倍;同时, CAT的活性变化则呈升高、降低、再升高、再降低4阶段,0.5mg/L时其活性达到最大值(p<0.01),除0.005mg/L和0.25mg/L浓度组CAT活性受抑制外,其余浓度组CAT的活性均高于对照组,由对照组的1.1倍到1.9倍,染毒浓度为0.25ms/L时, GST的活性达到最大诱导(p<0.01),其活性先缓慢上升后下降,并且各处理组GST的活性均显著高于对照组(p<0.05).在单一浓度TBBPA污染暴露的10d时间内,颤蚓体内SOD的活性变化趋势形成了一个变形的"M" 形曲线,在第3d时其活性达到最大诱导,而CAT活性的变化则表现为一个不规则的"N"形曲线,在第5d,时其活性达到最大诱导,并且SOD的活性比CAT的活性受时间的影响小,相对来说更稳定.可见, SOD与GST的活性变化似乎更能反映出TBBPA对颤蚓的污染效应及其毒性作用,而SOD的活性变化似乎更为灵敏,但是二者能否作为指示TBBPA污染的生物标志物尚需进一步研究.     

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.     

同时提取土壤微生物DNA和RNA方法的研究

获得高浓度、大片段、多样性程度高的土壤微生物总DNA是研究土壤微生物群落结构的分子生态学基础.研究采用PBS缓冲液洗涤土壤样品,结合SDS裂解微生物细胞的方法,同时提取分别添加小麦秸秆粉和油菜秸秆粉的两种土壤样晶的微生物DNA和RNA.对提取出的DNA和RNA进行酶切,基因组PCR和反转录PCR,实验结果表明该法提取的...     

Tyrosinase-modified carbon felt-based flow-biosensors: The role of ultra-sonication in shortening the enzyme immobilization time and improving the sensitivity for p-chlorophenol

Tyrosinase (TYR) was covalently immobilized onto amino-functionalized carbon felt surface via glutaraldehyde-coupling under ultrasonic treatment for 10 min. The resulting TYR-immobilized carbon felt was used as a working electrode unit of bioelectrocatalytic flow-through detector for TYR substrates (catechol, p-chlorophenol (p-CP), p-cresol, phenol etc.). Cathodic peak currents based on the electroreduction of enzymatically produced o-quinones were detected at –50 mV vs. Ag/AgCl. Compared with previous work in which TYR was immobilized onto amino-functionalized carbon felt for 16 hr without the ultrasonic treatment, we succeeded in (1) shortening the enzyme immobilization time from 16 hr to 10 min, (2) enhancing the sensitivity of p-CP, and (3) improving the operational stability of p-CP. The ultrasonic treatment during the TYR immobilization step would lead to certain changes in the structure of the immobilized TYR and the morphology of the immobilized TYR-layer on the carbon felt surface.     

高效氯氰菊酯对鲤鱼的生化指标影响研究

研究了高效氯氰菊酯对鲤鱼的脑、鳃、肝和骨骼肌肉中代谢酶、RNA和蛋白质的影响.用0.356 μg/L高效氯氰菊酯处理鲤鱼21d后,其脑、腮、肝、骨骼肌肉中6-磷酸葡萄糖脱氢酶(G-6-PDH)和乳酸脱氢酶(LDH)的活性逐渐降低,酶活性降低了35.2%～48.4%,RNA质量比减少了21.55%～45.65%,蛋白质质量比减少了41.72%～49.07%;撤离高效氯氰菊酯后,鲤鱼脑、鳃、肝和骨骼肌肉中的G-6-PDH和LDH酶活性、RNA和蛋白质质量比恢复到对照水平.结果表明:高效氯氰菊酯亚致死浓度处理后的鲤鱼脑、鳃、肝和骨骼肌肉中代谢酶活性、RNA和蛋白质质量比都逐渐下降,高效氯氰菊酯对鲤鱼的生化指标影响显著.