coarse isolation of siderophores secreted by marine microalgaes and new application of analytical techologies

在铁限制的条件下，海洋微型生物会释放一些低分子量并与铁有极强亲和力的配位体－铁载体。它可以在细胞外的环境中夺取Fe（Ⅲ），满足生物体对铁的需求。而铁对海洋生物的生长起着十分重要的作用。因此，铁载体的研究对于解释HNLC海区以及铁限制假说都具有十分重要的意义。基于此，本论文工作对微藻类分泌的铁载体进行了粗分离，探索了HPLC-MS联用技术用于铁载体与金属结合的分析以及共振瑞利散射法的含量测定技术。其意义在于为海洋生物可利用铁的形态研究打下铺垫，为分析技术开辟更广阔的应用领域。第一章通过大量文献，对铁载体的有关理论知识、研究意义以及国内外对铁载体的研究状况进行了归纳和总结。在此基础上，归纳了本论文...Under iron-limited conditions, marine microorganisms are able to secrete low-molecular-weight, highly specific iron chelators termed siderophores. These compounds scavenge iron from protein complexes or insoluble hydroxides in the extracellular environment and transport it to the cell cytosol via a membrane-bound receptor protein in the microorganism. Iron is a necessary requirement in marine ecos...学位：理学硕士院系专业：化学化工学院化学系_分析化学学号：20022502

Oil Spill identification by near-infrared spectroscopy coupled with clustering analysis based on principle components

鉴于目前没有一种方法能独立解决溢油鉴别的所有问题,本文提出了近红外光谱技术结合主成分聚类分析鉴别溢油种类的方法。通过有机溶剂萃取出自行配制的汽油、柴油和润滑油模拟样品中的溢油后记录其近红外光谱,对5800～6200cm-1区段范围内的谱图经多元散射校正(MSC)、Norris一阶导数平滑预处理处理后求其主成分,并在主成分的基础上引入Ward聚类分析法(离差平方和法)对样品分类。结果表明近红外光谱技术结合聚类分析能对体积分数在0.4～0.8mL/L间的海面溢油样品正确、快速分类,近红外光谱技术结合主成分聚类可作为溢油鉴别的一种辅助方法。Owing to none of the existing method can do it all for oil identification, but a method by near-infrared spectroscopy (NIR) coupled with clustering based on principle components is proposed. Petroleum oil stimulating samples of gasoline, diesel fuel and lubricating oil were employed to develop the method. NIR spectra ranges of 5800～6200cm~(-1) were recorded with the multiplicative signal correction and Norris first derivative filter pretreatments. The clustering analysis by Ward's method base on the different principle components was discussed. The results show that the method provides the rapid and accurate classification for spilled oil with concentration between 0.4～0.8 μL/L.国家高技术发展计划(863)资助项目(2001AA635040

Research on siderophore secreted by Prorocentrum donghaiense Lu under iron-limited conditions

在铁限制条件下,进行东海原甲藻分泌铁载体的动态研究。对藻类在富铁与缺铁条件下生长状况、生长过程中分泌铁载体的情况以及海藻接种量对铁载体分泌的影响进行了连续观测,结果表明:东海原甲藻在缺铁条件下生长状况远不如在富铁条件下;随着藻类的生长,分泌铁载体不断增多,达指数生长期时,其分泌量也达到了最大值,之后藻类的生长和铁载体分泌都呈现下降趋势;高接种量东海原甲藻能分泌较多的铁载体,并在较短时间到达峰值。Under iron-limited conditions,marine microorganisms are able to secrete low-molecular-weight,highly specific iron chelators termed siderophores.These compounds scavenge iron from protein complexes or insoluble hydroxides in the extracellular environment and transport it to the cell cytosol via a membrane-bound receptor protein in the microorganism.Iron is a necessary requirement in marine ecosystems.Therefore research on siderophores is very significant to interpretation of high nitrate low chlorophyll(HNLC) regions and iron hypothesis.However,it is not clear whether eukaryotic phytoplankton can produce the iron-specific chelating compounds.Therefore,a systematic investigation was carried out for siderophore secreted by P.donghaiense Lu under iron-limited conditions.Parameters studied included the growth of the algae in iron-added and iron-deficient medium,the secretion of siderophore during algae growth,and the effect of algae population inoculated on the siderophore production.Results showed that the algae growth rate in iron-deficient medium was slower than those in iron-rich medium.Under iron-limited conditions,the siderophore production increased along with algae growth.Maximum production was observed in the exponential growth period.Thereafter the rate of algae growth and siderophore secretion started to decrease.Furthermore,increasing algae populationt resulted in secreting more siderophore and shortening the time needed to reach maximum production.国家高技术研究发展计划资助项目(2001AA635040);; 国家海洋局青年基金资助项目(2005102);; 国家重点基础研究发展规划资助项目(2001CB409702);; 青岛市科技将材计划资助项目~

(131)~I标记卵清蛋白及其载(131)~I-OVA的PLGA/DDAB纳米粒SPECT显像

制备载(131)~I-OVA的荷正电聚乳酸-乙醇酸(PLGA/DDAB)纳米粒,探讨其在活体动物体内的生物分布与转运研究。氯胺T法对卵清蛋白(OVA)进行(131)~I标记,Sephadex G 25纯化标记抗原,纸层析法测定标记物的生物活性和体外稳定性。纳米沉淀法制备PLGA/DDAB纳米球,将其与(131)~I-OVA共混吸附,制备载(131)~I-OVA的PLGA/DDAB纳米粒。动物试验选择C57/BL6小鼠,肌肉注射载(131)~I-OVA的PLGA/DDAB纳米粒,分析(131)~I-OVA在小鼠组织器官中的分布情况,并于注射疫苗制剂后不同时间行SPECT正位静态显像,观察小鼠不同组织内放射性浓聚情况。结果显示,标记抗原经纯化后,测得(131)~I-OVA比活度达32~42&mu;Ci&middot;&mu;g-1,标记率达(71.92&plusmn;0.08)%,放射性化学纯度为(85.94&plusmn;0.15)%。标记物在新鲜血清中存放72 h后,仍保持很高的反应活性,表明标记的OVA稳定性良好。采用纳米沉淀法制备PLGA/DDAB纳米球,平均粒径、分散系数和Zeta电位分别为122.8 nm、0.084和+31.5 m V。将其与(131)~I-OVA共混吸附,抗原携载率达(89.60&plusmn;0.21)%。注射载(131)~I-OVA的PLGA/DDAB纳米球疫苗制剂,与无佐剂疫苗相比,抗原在注射部位消失的速度更为缓慢。结果表明,成功制备了载(131)~I-OVA的PLGA/DDAB纳米球,生物活性较好,可用于抗原在活体动物的生物分布与转运研究。</p

Applications of Analytical Technology in Studying Siderophore From Marine Phytoplankton

铁对海洋浮游植物的生长起着重要的作用。在高营养盐低叶绿素(HNLC)海区,铁是藻类生长的限制因子;而在近岸,铁又是赤潮发生的激发因子之一。研究表明,可溶性铁对浮游植物生长有很强的控制作用,大部分可溶性铁是同具有极强铁亲和力的配位体螯合在一起的,如铁载体。因此,在对铁限制、铁加富实验的研究中,铁载体的提取与分析技术显得尤为重要。着重介绍了海洋浮游植物中铁载体的提取及检测分析技术。Iron plays an important role in the growth of marine phytoplankton. In the high nutrient and low chlorophyll sea area, iron is a limiting factor of algal growth, and in the nearshore area, iron is one of factors to stimulate the occurrence of red tide. It is shown from study results that the soluble iron has a strong controlling effect on the phytoplanktonic growth, and most of soluble iron is chelated with ligand such as siderophore with high affinity to Fe 3+ . Therefore, the extraction and analysis technology of side- rophore is of great importance to the iron-limiting and iron-enrichment experiment and research. In this paper, the extraction, assay and analysis techniques of siderophore from marine phytoplankton are introduced in detail.国家高技术研究发展计划项目——海洋石油污染物现场实时监测的光纤近红外传感仪器系统(2001AA63-5040