极大螺旋藻铁型超氧物歧化酶的纯化及性质

极大螺旋藻 Ｓｐｉｒｕｌｉｎａｍａｘｉｍａ藻体ＳＯＤ聚丙烯酰胺梯度凝胶电泳呈现４条同工 酶带, Ｈ_ ２Ｏ_２明显抑制该酶活性, ＫＣＮ对酶活性无影响,确认为 Ｆｅ－ＳＯＤ经硫酸铵分部盐析、离子交 换柱层析及凝胶过滤,纯化到电泳单斑点均－程度．纯化的Ｆｅ－ＳＯＤ分子量为３９．３ＫＤ,亚基分子 量为２０ＫＤ金属元素分析表明,每个亚基含０ ５５个Ｆｅ原子,该酶在紫外区最大吸收峰值为 ２７５．８nｍ、该酶氨基酸组成与蓝、绿藻和高等植物的Ｆｅ－ＳＯＤ相似,但它含有较高的丙氨酸,酸性氨 基酸和碱性氨基酸比值与低等植物及原核生物相近而明显高于高等植物．     

利用湖泊内源性饵料饲喂中华绒螯蟹(Eriocheir sinensis)的养殖与净水效应

在东太湖网围养殖区进行不同放养密度(3000、6000、9000只/hm²)饲喂太湖水草与螺蛳的中华绒螯蟹(Eriocheir sinensis)养殖实验,各处理组均设投喂鱼肉与玉米等外源性饵料的对照,以获得不同养殖模式对中华绒螯蟹生长、品质及养殖水环境的影响,以及利用浅水草型湖泊内源性饵料养殖中华绒螯蟹的适宜放养密度.结果表明,规格和增肉倍数虽然均显著小于相同养殖密度下投喂外源性饵料的对照组,但是各实验组商品蟹体重均能达到150 g;低密度养殖组(3000只/hm²)的产量与对照组无显著差异,其余两组显著低于对照组;而低密度养殖组的河蟹回捕率显著低于对照组,其余两组则无显著差异.实验养殖区水质状况优于投喂外源性饵料的对照区,其TP含量显著低于对照区,投喂外源性饵料会增加水体N、P含量,导致水体富营养化.投喂太湖水草与螺蛳等内源性饵料进行中华绒螯蟹养殖不但可以有效地抑制浅水湖泊水草的疯长,还可通过中华绒螯蟹收获从水体输出N、P营养盐,起到净化水质的作用.不同密度养殖的生态与经济效益对比结果显示,利用草型湖泊内源性饵料养殖中华绒螯蟹的适宜放养密度为6000只/hm²左右.     

提高地浸钻孔出水量的有效技术措施

地浸钻孔是一种以采矿为目的的钻孔,提高钻孔出水量就是提高钻孔的采矿量。通过对多年地浸钻孔施工经验和理论进行分析,发现影响钻孔出水量的主要因素为钻井泥浆的成分和性能以及人工过滤层的渗透性差异,提出在地浸钻孔施工方案中采用植物胶净化泥浆钻进和扩孔、投砾罐射吸式填砾、物理化学组合洗井3种技术措施,对比前后施工钻孔的出水量大小,证明了该技术措施对提高地浸钻孔出水量具有良好的效果。     

Nitrifying and Denitrifying Bacteria in Epiphytic Communities of Submerged Macrophytes in a Treated Sewage Channel

A quantification of nitrifying and denitrifying bacteria present in different compartments (water, sediments, submerged macrophytes) of a treated sewage channel was made to estimate their influence on the nitrogen balance and to assess the significance of macrophytes for nitrification and nitrogen conversions in general. Considerable numbers of autotrophic and heterotrophic nitrifying and denitrifiying bacteria were found to be present in the epiphytic communities of different species of submerged macrophytes of a treated sewage channel. Comparing the influence of the different compartments on total stream nitrification and denitrification it could be concluded that dense beds of submerged macrophytes particularly positively influence nitrification. Epiphytic nitrifiers were estimated to be as important for the total nitrification as nitrifiers in the sediment. Denitrification was mainly taking place in the sediment. The influence of the suspended nitrifiers and denitrifiers on the nitrogen balance was assumed to be negligible.     

内蒙产高烧失低品位硅藻土的提纯及碳化性能

针对内蒙产高烧失低品位硅藻土的提纯,采用水洗[CD*2]焙烧工艺进行了实验。探索了水选次数、焙烧条件等对提纯效果的影响。采用XRD、FTIR、SEM、BET和化学分析等方法对提纯试验获得的产物进行了表征。结果表明:采用水选法可以实现石英、长石和黏土类杂质的去除,有机质的去除采用600 ℃焙烧就可以获得理想的提纯效果。提纯硅藻土SiO₂质量分数达到82.98%以上,比表面积可达44.5 m²/g,堆积体积达21 mL/10g。在隔绝空气条件下焙烧水选提纯土,获得的碳化硅藻土与橡胶相容性好,可替代半补强炭黑。     

海带中抑藻活性化合物的分离及抑藻活性分析

以2000 g海带为原料、米氏凯伦藻为测试微藻,采用甲醇浸提、液液萃取、硅胶柱层析、Seph-adax LH-20凝胶柱层析和硅胶薄层层析等分离方法,纯化可能具有抑藻活性的化合物,制备到5个薄层纯样品,B21(1.39 g)、B131(0.51 g)、B132(0.48 g)、C211(0.016 g)和C212(0....     

Optimization of the purification methods for recovery of recombinant growth hormone from Paralichthys olivaceus

This study aimed to optimize the purification of recombinant growth hormone from Paralichthys olivaceus. Recombinant flounder growth hormone (r-fGH) was expressed by Escherichia coli in form of inclusion body or as soluble protein under different inducing conditions. The inclusion body was renatured using two recovery methods, i.e., dilution and dialysis. Thereafter, the refolded protein was purified by Glutathione Sepharase 4B affinity chromatography and r-fGH was obtained by cleavage of thrombin. For soluble products, r-fGH was directly purified from the lysates by Glutathione Sepharase 4B affinity chromatography. ELISA-receptor assay demonstrated that despite its low receptor binding activity, the r-fGH purified from refolded inclusion body had a higher yield (2.605 mgL-1) than that from soluble protein (1.964 mgL-1). Of the tested recovery methods, addition of renaturing buffer (pH 8.5) into denatured inclusion body yielded the best recovery rate (17.9%). This work provided an optimized purification method for high recovery of r-fGH, thus contributing to the application of r-fGH to aquaculture.     

Preparation of hemocyanin polyclonal antibody and the identification of 35 kDa hemocyanin fragment in HaHotis diversicolor Reeve

以杂色鲍（Haliotis diversicolor Reeve）为研究对象,通过密度梯度离心方法纯化得到高纯度血蓝蛋白,以其作为抗原皮下注射免疫新西兰大白兔,从而获得高效价的兔源多克隆抗血清。进一步通过ProteinA抗原亲和纯化的方法对该抗血清纯化,最终获得效价更高、检测特异性更好的血蓝蛋白多克隆抗体。应用该抗体进行Western检测发现,鲍血淋巴中存在着多样的血蓝蛋白衍生产物;进一步结合质谱技术对其中35kDa条带进行鉴定发现,其来源于血蓝蛋白I型亚基的H结构域。     

菲律宾蛤仔碱性磷酸酶的分离纯化及部分性质研究

经Tris-HCl缓冲液浸提、硫酸铵分级沉淀盐析、DEAE-Sepharose Fast Flow离子交换层析和Sephadex G-75凝胶过滤层析等步骤,从菲律宾蛤仔内脏团中分离得到碱性磷酸酶,SDS-PAGE显示为单一条带,提纯倍数为14.42,比活力达到131.08U/mg。酶学性质研究表明:该酶亚基分子量约44kD,最适反应温度为45℃,最适pH值为9.0,米氏常数Km为0.098mmol/L,最大反应速度Vmax为1.01μmol/(mL·min)。Ca2+、Mg2+对酶活力表现出显著的激活作用。     

红笛鲷NCCRP-1基因原核表达条件的优化及纯化

通过克隆编码红笛鲷(Lutjanus sanguineus)非特异性毒性细胞受体蛋白-1(nonspecific cytotoxic cell receptor protein-1,NCCRP-1)成熟肽基因序列,然后与载体连接构建pET21a-NCCRP融合蛋白表达载体,将其转入大肠杆菌BL21进行诱导表达并优化条件。SDS-PAGE分析表明,在异丙基-β-D-硫代半乳糖苷(IPTG)浓度0.8 mmol/L、37℃条件下培养3 h后表达量最大,分子大小与预期值相符,融合蛋白主要以包涵体形式高效表达,通过HisTrap HP柱子使其得到进一步纯化;Western blot分析表明,该融合蛋白可与鼠抗His-tag单克隆抗体发生特异性结合,说明获得该表达产物。