5 resultados para Chlorocatechol dioxygenase

em Chinese Academy of Sciences Institutional Repositories Grid Portal


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Bone marrow-derived mesenchymal stem cells (MSCs) hold great promise for treating immune disorders because of their immunoregulatory capacity, but the mechanism remains controversial. As we show here, the mechanism of MSC-mediated immunosuppression varies

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The dibenzofuran (DF)-degrading bacterium, Janibacter terrae strain XJ-1, was isolated from sediment from East Lake in Wuhan, China. This strain grows aerobically on DF as the sole source of carbon and energy; it has a doubling time of 12 hours at 30 degrees C; and it almost completely degraded 100 mg/L-1 DF in 5 days, producing 2,2',3-trihydroxybiphenyl, salicylic acid, gentisic acid, and other metabolites. The dbdA (DF dioxygenase) gene cluster in the strain is almost identical to that on a large plasmid in Terrabacter sp. YK3. Unlike Janibacter sp. strain YY-1, XJ-1 accumulates gentisic acid rather than catechol as a final product of DF degradation.

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随着人类基因组计划的完成,蛋白质组学研究已成为生命科学乃至自然科学中最活跃的学科领域之一。蛋白质组学将对我们理解后基因组学时代的基因功能有极大的帮助。差异表达蛋白质组学通过对不同样品蛋白质表达水平的比较,已经被广泛地应用在疾病的研究领域。P糖蛋白已经被证实参与介导了在急性粒白血病中发生的多药耐药,然而越来越多的研究结果表明,在急性粒白血病中多药耐药的机制是多因素的。为了寻找白血病肿瘤细胞中与耐药相关的靶蛋白,我们采用蛋白质组学研究技术,对多药耐药细胞系K562/A02的蛋白质表达谱进行了分析。 本研究采用蛋白质组学研究技术,对多药耐药细胞系K562/A02和药物敏感细胞系K562/S的总蛋白质进行双向凝胶电泳(2 DE)的差异分析,结果发现在K562/A02细胞中有41个蛋白质点差异显著,并用基质辅助激光解离飞行时间质谱(MALDI-MS)成功鉴定了大多数的蛋自点。在被鉴定出的差异蛋白质中,有一些是首次发现与耐药相关,包括chloride intracellular charmel,chaperonin,proteindisulfide-isomerase ER6o precursor and constitutive heat shock protein 70等。所得结果不但能够使我们更加深入的理解肿瘤耐药的病理机制,更重要的是通过找到肿瘤细胞发生耐药的标志性蛋白,可以为开发肿瘤耐药预测方法和寻找抗耐药肿瘤靶标新药的线索,同时也为计算机虚拟筛选设计抗耐药肿瘤新药奠定基础。除了详细的研究了K562/A02细胞系差异蛋白质组,我们还采用蛋白质组学的方法,以乳鼠心肌细胞为实验模型,研究在ARs激动的不同清况下心肌细胞蛋白质表达谱的变化。心肌肥厚是心脏工作超负荷的一种代偿反应。目前,越来越多的证据表明ARs在心肌肥厚的发生中起着非常重要的作用,但人们对其机制了解尚不全面。通过检测心肌肥厚时全蛋白表达的变化,将有助于我们全面、深入了解ARs在心肌肥厚中的作用机制。通过严格的2D胶图象分析,蛋白取点后胶内水解,高灵敏度的质谱鉴定及数据库检索,我们成功地进行了乳鼠心肌细胞肥厚为模型的蛋白质组学的研究。本实验通过比较研究不同肾上腺素受体激动剂norePinephrine伽E),phenylephxine(PE)and isoprenaline(ISO)诱导组与未诱导对照组的蛋白质组学变化,分析确定蛋白丰度差异在2倍以上的蛋白质点,并通过MALDI-MS进行蛋白鉴定。20个具有显著差异的蛋白质被鉴定,包括caspase-11,vimeniin,4-hydroxyphenylpyruvic acid dioxygenase,Modl protein,Txndcs protein等,其中一些蛋白在此之前并没发现与心肌肥厚相关,对它们在心肌肥厚中的意义和机制我们将做进一步的探讨。本研究结果将为我们更好理解肾上腺素受体之间以及肾上腺素受体与心肌肥厚之间的关系提供有益的线索,并将为最终发现可应用于临床的新的诊断和治疗心血管疾病的标志性蛋白打下基础。

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沙棘广泛分布于亚欧大陆的温带地区和亚洲亚热带的高海拔地区。沙棘能适应多种生态环境,能耐受多种逆境(如干旱、低温、高温和盐害等)。在中国,沙棘常常被用作植被恢复中的先锋树种而大量栽培。本文以中国沙棘为试验材料,探索沙棘适应干旱机制,以及沙棘对干旱胁迫的适应机制是否存在种群间的差异,同时试图通过分析干旱胁迫下沙棘叶片蛋白质表达变化探索沙棘适应干旱胁迫的分子机理。 对三个分别来自低海拔湿润地区、低海拔干旱地区和高海拔湿润地区的中国沙棘种群进行干旱胁迫处理。干旱胁迫能提高根冠比,比叶面积,降低平均叶面积和总生物量,提高沙棘的抗氧化性酶活性、脯氨酸含量、脱落酸(ABA)含量、降低光合作用,提高长期用水效率。实验中的这两个低海拔种群比高海拔种群抵抗干旱的能力更强,不同的种群采用了不同的策略来耐受干旱胁迫和过氧化胁迫。 在2004 年度的实验中,干旱胁迫处理下,高海拔湿润种群(道孚种群)严重失水,生长也受到更大的抑制,非气孔因素在抑制光合作用方面占支配地位,抗坏血酸含量下降,ABA和脯氨酸含量增加幅度比九寨沟种群的要高,这可能是因为道孚种群严重失水而引起的;而低海拔湿润种群(九寨沟种群)的体内水分状况几乎不受干旱的影响,生长情况也较道孚种群要好。 在2005 年度的试验中,和高海拔湿润地区种群(道孚)相比较,低海拔干旱地区种群(定西)在叶片相对水含量、根冠比、抗氧化酶活性(过氧化氢酶、抗坏血酸过氧化物酶和谷胱甘肽过氧化物酶)、保护性物质(脯氨酸,脱落酸)含量等方面都要高,光能热耗散能力也更强,而且气体交换参数(气孔扩散阻力和胞间CO2浓度等)对干旱也更不敏感。 分析了干旱胁迫下沙棘叶片蛋白质表达的变化。共发现319 个蛋白质,有4 个蛋白在干旱胁迫下消失(Putative ABCtransporter ATP-binding protein 、Hypothetical proteinXP-515578,热激蛋白Hslu219 和一个没得到鉴定的蛋白),4 个只在干旱胁迫下出现(没命名的蛋白质产物,对甲基苯-丙酮酸双加氧酶,NTrX 和一个没得到鉴定的蛋白),46 个蛋白质的表达丰度变化显著,包括32 个干旱负调蛋白,14 个干旱正调蛋白(3 个Rubisco 的大亚基、J-type–co-chaperone Hsc20、putative protein DSM3645-2335、putative acyl-COA 脱氢酶、nesprin-2 和两个没有得到鉴定的蛋白质)。这些蛋白质参与了氮代谢调控、抗氧化行物质的合成、脂肪酸β-氧化、核骨架构造、[Fe-S]基团组装、物质跨膜运输、细胞分裂或作为分子伴侣和蛋白质酶起作用。putative ABC transporter ATP-binging protein、NtrX、nesprin-2 和Hslu 是本试验新发现的高等植物蛋白,我们主要从它们的保守结构域或在其他生物中的同源物来猜测它们的功能。实验结果为我们研究植物抗干旱机制提供了新线索和新视野。 Seabuckthorn (Hippophae rhamnoides L.) is widly distributed throughtout the temperatureresiogn of Europe and Asia and sub-tropical plateau zone of Asia. H. rhamnoides can adapatseveral different environments, and can tolerant several envioronmental stresses (e.g, lowtemperature, high temperature, drought and salty). It has been widely used in forest restoration asthe pioneer species in China. In present study, we applied H.rhamnoides subsp. Sinensis asexperimental materials to study its drought-tolerant mechanism, and expected to findpopulational difference in drought-tolerant mechanism that may exist among populations, and tryto get some insight in drought-tolerant mechanism of it at morecular level through analyzing thechange of leaf protein expression. Three populations from high altitude wet zone, low altitude wet zone and low altitude arid znoe,respectively, were applied in our experiment, and were subjected to drought. Drought increasedthe root/shoot ratio(RS), special leaf area, long-term water use efficinency, activity of antioxidantenzymes, proline content and abscisic acid (ABA) content, declined the net photosynthesis rate(A), average leaf area (ALA), total biomass (TB). Both two low altitude populations were moredrought-tolerant than the high altitude population, and different population applied differentstratedgies to tolerant oxidant stress and drought stress. The results of the exprement in 2004 showed that Daofu population was more drought-sensitivethan Jiuzhai population. Under drought conditions, leaf relative water content (RWC) greatlydecreased in Daofu population, but not in Jiuzhai population. The large loss of water in Daofupopulation resulted in a limitation on A mainly caused by non-stomatal factors, severer suppression in growth rate and a significant reduction in ascorbic acid (AsA) content, comparedwith Jiuzhai population. The greater increase in content of ABA and proline in Daofu populationmay be also induced by large loss in water, so that enable plants to cope with sever drought. In the exprement of 2005, drought significantly increased RS, activities of catalase (CAT),peroxidase (POD), glutathione peroxidase (GPX) and ascorbate peroxidase (APX), and alsosignificantly increased ABA and proline contents. On the other hand, compared with Daofupopulation, drought induced larger RS and activities of CAT, GPX and APX, and higher ABAcontent in Dingxi population, whereas gas exchange traits, e.g., stomatal limitation value (LS) andintercellular CO2 concentration (Ci), were less responsive to drought in Dingxi population thanthose in Daofu population. All these factors enable Dingxi population to tolerant drought betterthan Daofu population. The leaf protein profile of seabuchthorn subjected to drought was analyzed. Altogether 319proteins were detected in well-watered sample, four proteins disappeard by drought (putativeABCtransporter ATP-binding protein, hypothetical protein XP-515578, Hslu219and aunidentified protein), four only appeared under drought (a probable nitrogen regulation protein(NtrX), a 4-hydroxyphenylpyruvate dioxygenase , an unnamed protein product and an identified protein), 32 drought down-regulated proteins, and 14 drought up-regulated proteins (nine wereidentified: three large subunits of Rubisco, a hypothetical protein DSM3645-23351, a putativeacyl-COA dehydrogenase, a nesprin-2, a J-type-co-chaperone HSC20 and two unmatchedproteins). These proteins may involve in β-oxidation, cross-membrane transport, cell division,cytoskeleton stabilization, iron-sulfur cluster assembly, nitrogen metabolism regulation andantioxidant substance biosynthesis or function as molecular chaperone or protease. Four proteins(a putative ABC transporter ATP-binging protein, NtrX, nesprin-2, Hslu) were new found in highplants, and their functions were estimated from their conserved domain or their homologues inother organism. Our results provided new clue and new insight for us to study thedrought-tolerant mechanism in plants.

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近年来,我国炼油行业发展迅速,炼油能力全世界第二,炼油行业已成为污染大户。本研究针对炼油废水生物处理中存在的稳定达标难、抗冲击负荷能力差、建设投资与运行成本高等问题,就菌剂强化处理炼油废水中试与工程应用展开了研究,以期为菌剂的工程应用与推广提供理论参考与技术支持;并以炼油废水中的主要特征污染物苯酚为研究对象,考察了不同浓度苯酚冲击下功能菌的响应机制,并以此为指导研制功能菌激活促进剂,考察其对功能菌生物学指标的调控效果,以期为废水生物处理有毒污染物冲击调控提供理论依据与技术支持。 中试研究表明,菌剂强化处理炼油废水,出水COD、NH4+-N 平均值为86.7、7.6 mg/L,其平均去除率较常规生物处理系统分别提高了35.47%、59.28%,其耐受COD、NH4+-N 容积负荷分别高达2.42、0.139kg/(m3·d),具有良好的耐冲击能力。工程应用研究表明,菌剂强化处理炼油废水,出水COD、NH4+-N 平均值分别为85.05、8.4mg/L,其去除率较常规生物处理系统提高了25.1%、28.7%,出水水质各项指标均达到了国家《污水综合排放标准GB 8978-1996》一级排放标准。技术经济分析表明,菌剂强化处理炼油废水在建设成本、运行成本上分别降低38%、49%,具有良好的技术经济优势。 苯酚冲击下功能菌响应机制研究表明:不同浓度苯酚冲击下,生物学指标生物量、脱氢酶酶活、1,2-双加氧酶酶活对冲击都有不同程度的响应,其响应敏感程度为脱氢酶酶活>生物量>1,2-双加氧酶酶活。1,2-双加氧酶酶活与COD 降解率相关性良好,可表征苯酚降解过程,确认为调控重点。以此为指导研制出苯酚降解功能菌抗冲击激活促进剂,可有效调控功能菌对有毒污染物苯酚的降解效果,1000mg/L 苯酚冲击下,经调控,其COD 去除率较对照提高20%,降解时间缩短16%以上。其对生物学指标的调控效果为1,2-双加氧酶酶活>生物量>脱氢酶酶活,验证了功能菌在苯酚冲击下的响应机制。研究表明菌剂强化处理炼油废水切实可行,具有良好的技术经济优势。有毒污染物冲击下废水生物处理系统响应机制研究为抗冲击调控提供了新的研究思路。 Currently, China’s oil refining industry is developing rapidly and has become the second largest all over the world. The oil refining industry is one of the major pollution industries in our country. The pilot scale study and engineering application research were conducted aiming at the problems in refining wastewater such as poor treatment stability and water quality, poor anti-shock capacity and expensive running cost, etc., so as to provide theoretical references and technological supports for the engineering application and popularization of microbial preparation in wastewater treatment. Also, the response mechanism of functional microbe under shock of different phenol concentrations, which is the main pollutants in refinery wastewater, was studied. Based on this result, functional microbe activation accelerator was developed, and the regulation effect of functional microbe biological index under phenol shocking were studied, in order to provide theoretical basis and technological support for regulation of toxic shocking of wastewater biological treatment. The result of pilot scale research indicated: for treatment of refinery wastewater in bioaugmention treatment system of microbial preparation, the COD and NH4+-N average value of effluent was 86.7 and 7.6 mg/L, Comparing with normal biological treatment system, the average removal rates of COD, NH4+-N increased 35.47%,59.28% separately by bioaugmention treatment system, which showed better anti-shocking capacity, the volumetric load r of COD and NH4+-N reached 2.42 kg/(m3·d) and 0.139 kg/(m3·d), respectively. The research on engineering application of refinery wastewater bioaugmentation treatment by microbial preparation indicated:the average concentrations of effluent COD and NH4+-N in the bioaugmentation treatment system were 85.05 and 8.4mg/L, which increased by 25.1% and 28.7% comparing with normal biological treatment system of refinery wastewater, And the effluent quality meets the first grade of discharging standard of National Integrated Wastewater Discharge Standard GB 8978-1996. The economic analysis of technology indicated: the demonstration project of bioaugmentation treatment of refinery wastewater by microbial preparation decreased by 38% in construction cost and 49% in running cost. This technology has economic benefits. The response mechanism of functional microbe under phenol shock indicated: biological index such as the biomass concentration, dehydrogenase and 1,2-dioxygenase had different responses under phenol shocking of different concentrations. The response sensitivity of different biological index under phenol shocking of different concentrations is: dehydogenase activity > biomass >1,2-dioxygenase activity, and high correlation of 1,2-dioxygenase and COD degradation percentage is achieved, thus 1,2-dioxygenase could be used to reflect the degradation situation of pollutants. So, 1,2-dioxygenase is the keypoint of regulation. The anti-shock activation accelerator of phenol degradation functional microbe was primarily developed. The results indicated: the activation accelerator could regulate the degradation effect of toxic substance-phenol by functional microbe effectively. For the functional microbe treatment system under phenol shocking of 1000mg/L, the COD degradation rate increased by 20% and the degradation time reduced by more than 16% under regulation of activation accelerator. The regulation effects of biological index are: 1,2-dioxygenase > biomass > dehydrogenase. In this way, the response mechanism of functional microbe under toxic shocking is verified. The result indicated: the augmented microbial preparation treatment of refinery wastewater is applicable. It has many technical and economical advantages. The research results of responses mechanism of wastewater treatment system on toxic pollutants would offer a new idea for regulation of anti-shock.