L-异亮氨酸高产菌定向育种与代谢控制发酵

以L-异亮氨酸产生菌A_(41-3)为出发菌株，经过定向育种和与L-缬氨酸生物合成突变株原生质体融合，从中选育出一株抗α-氨基-β-羟基戊酸、S-（-2-氨基乙基-）-半胱氨酸、乙硫氨酸、异亮氨酸羟肟酸、2-噻唑丙氨酸和红霉素的亮氨酸生物合成缺陷型CN_(69-1)，通过对L-异亮氨酸生物合成代谢控制发酵条件的研究，在含葡萄糖11%，生物素100 μg/L，硫酸铵4.5%，磷酸二氢钾0.3%，硫酸镁0.02%，硫胺素800 μg/L，硫酸亚铁0.004%，硫酸锰0.004%，碳酸钙2%，L-亮氨酸0.02%，pH6.8-7.0培养基中，32 ℃摇瓶发酵72h，可产生L-异亮氨酸23.6g/L。通过定向筛选缬氨酸代谢活性菌株CN_(69-8)，使缬氨酸生物合成量由8.73g/L下降至1.81g/L，L-异亮氨酸生物合成量保持在24g/L左右。在2.000L发酵罐上进行了pH值、通气量、补料等培养条件优化的研究，在适宜条件下，菌株CN_(69-8)发酵44h，L-异亮氨酸生物合成量可达20g/L以上。测定了有关菌株L-异亮氨酸生物合成关键酶苏氨酸脱氢酶、乙酰羟酸合成酶和分枝链氨基酸转氨酶性变化，结果表明，有关关键酶的活性均比出发菌株为高。通过生长谱法确定L-亮氨酸生物合成途径所缺失的酶是异丙基苹果酸脱氢酶。采用732阳离子交换树脂从发酵液中分离提取L-异亮氨酸。研究了发酵液酸化pH值、吸附体积、不同浓度洗脱剂以及单柱吸咐与双柱串联吸咐对L-异亮氨酸分离提取的影响。确定酸化pH值为2.0，以双柱串联吸咐，用0.2mol.L~(-1)氯化铵-0.1 mol.L~(-1)氨水复合洗脱分离的工艺条件，提取总收率可达45%以上。发酵产品经红外光谱，纸层析，比旋光度等项测定证明明是L-异亮氨酸。发酵中试分离提取的总收率可达40%以上，产品经检验确定为L-异亮氨酸，其质量符合《中华人民共和国药典》标准。

石羊河下游民勤绿洲景观变化与生态安全研究

民勤绿洲位于我国西北干旱区内陆河石羊河流域的下游。近些年来，随着该流域中、上游地区大量截留水资源，使进入民勤绿洲的地表水资源量逐年减少，导致绿洲生态环境不断恶化，生态安全面临着严重危机。本论文应用景观生态学原理与方法，综合运用3S技术，通过多时相遥感数据的对比分析，系统研究了民勤绿洲景观格局动态演变过程、形成原因及其与重要生态过程的相互作用，并在此基础上对绿洲生态安全的时空变化进行了综合评价。主要研究成果：1．景观格局影响生态过程，研究景观格局可以揭示生态过程变化的内在原因。因此，景观空间格局研究一直以来是景观生态学研究的核心与热点之一。应用景观空间格局分析软件FRAGSTATS对绿洲近14年的景观格局及其变化过程的定量研究表明，开荒与弃耕始终存在于景观的演变过程中，导致景观中林地、荒草地、耕地与沙地斑块类型的相互转化。开荒作用主要发生在1987-1998年间并导致林地与荒草地斑块转化成耕地斑块，绿洲耕地面积在此时期逐渐增加；弃耕作用主要发生在1998年至2001年间并导致耕地斑块逐渐转化成沙地斑块或荒草地斑块，耕地面积在此时期逐渐减少014年来，绿洲沙漠化过程逐年发展，主要表现为林地与荒草地斑块与耕地斑块逐渐转变成沙地斑块，导致沙地斑块面积（CA）逐渐增加，而斑块数量（NP）逐渐减少，使得沙地平均斑块大小（MPS）逐渐增大，沙地斑块呈现集中连片式的分布趋势。由于林草地斑块的逐渐减少以及耕地与沙地斑块的增加导致绿洲的多样性（SHOI）、均匀性（SHEI）与平行与并列指数（IJI）逐渐降低，而连通性（CONGTAG）逐渐增强，绿洲景观格局越来越强烈地受到沙地与耕地斑块类型的控制。绿洲景观的连通性上升的幅度（CONTAG）是坝区＞泉山区＞湖区。景观的多样性（SHDI）与均匀性（SHEI）下降的幅度是坝区＞泉山区＞湖区。坝区景观的衰退过程（沙地面积的增幅）与人类对该灌区的开发强度（耕地面积的增幅）与都强于泉山区与湖区。2．景观变化的主要驱动因素包括自然、社会与经济因素。绿洲景观格局的演变是以上因素综合作用的结果。干早区绿洲的形成机制决定了其发展与演变强烈地受到水资源的质与量的支配，水资源条件的变化是驱动该绿洲景观演变的主导因素。由于中、上游地区为满足经济发展的需要，大量截留地表水资源使进入下游民勤绿洲的地表水资源从50年代的年平均5.42亿m3锐减到2001年的0.73亿m3，致使绿洲地下水的补给量减少，而耕地面积的扩大导致地下水的超采量增加，引发土地干旱化与盐碱化，大量沙生植被枯萎、死亡，弃耕地增加。民勤绿洲用水模式进步缓慢、管理效率低下、人口的快速增长、农业技术推广速度缓慢以及土地使用制度等因素共同作用形成了推动景观变化的复杂人文机制。水资源的减少与人文机制共同作用驱动着绿洲景观的演变过程。3．指出绿洲景观的生态安全应包括景观的格局安全与过程安全两个方面。景观格局与生态过程密不可分，绿洲景观格局影响生态过程，生态过程“塑造”景观格局。利用ARCv正W的空间分析功能，从景观格局的空间邻接特征角度研究景观斑块类型受沙漠化威胁与盐碱化威胁的程度，并在此基础上分析生态安全。结果表明坝区耕地、林地、荒草地受沙漠化威胁程度高于其它两个灌区，湖区的耕地、林地、草地受盐碱化威胁的程度高于其它两个灌区。水对干旱区绿洲的重要性决定了水生态过程研究是绿洲生态安全研究的重要方面。利用ARCMAP的空间插值的地统计分析功能对绿洲的地下水埋深与地下水矿化度进行空间插值，得到地下水埋深与地下水矿化度的时空分布，结果表明：各时期绿洲地下水埋深为中心深，边缘浅，表明地下水降落漏斗的存在与不断加深。14年来地下水位的平均下降速率，坝区最快，泉山区次之，湖区最慢，三灌区分别为0.75m、0.7m与0.60m。绿洲北部地下水矿化度显著升高，14年来泉山区地下水矿化度上升为1.36倍，湖区南部地下水矿化度上升1.47倍，湖区北部地下水矿化度上升为1.43倍，地下水矿化度平均值达到5.6g/L，成为绿洲地下水矿化度最高地带，也是作物产量锐减的地带，因此地下水矿化度的上升导致作物安全生长潜在格局发生变化，作物安全生长区面积缩小，而中度以上不安全生长区面积扩大，造成了作物的减产与农业产业结构调整的空间缩小，难度加大。4．绿洲为水、土、植被、气候、社会经济与人类活动等众多方面所组成的、彼此相互联系的复杂的社会一经济一生态复合系统。因此绿洲的生态安全的综合评价涉及吸素众多，必须采用定量与定性相结合的方法进行科学地评价。采用目前较为成熟的环境综合评价方法一层次分析法AHP，并构建绿洲景观生态安全评价指标体系，对绿洲景观生态安全进行综合评价，结果表明：绿洲及各灌区生态安全水平逐年降低，目前生态安全值与1987年的相比，绿洲为68％，坝区73.7％，泉山区为63.6％，湖区为65.2％。三灌区中坝区的生态安全水平在各时期都是最高的，其次为泉山区，湖区最低。14年间生态安全下降幅度较大的为泉山区与湖区，坝区生态安全水平下降幅度较小。5．将绿洲生态环境恶化所导致的经济损失值量化，运用市场价值法与机会成本法对近14年来由于绿洲的环境恶化导致对社会经济发展所产生的不良后果从植被死亡、土地盐渍化导致作物减产、弃耕地出现导致的土地生产力的丧失与地下水位下降所导致的机井报废与提水成本增加四个方面进行经济效益损失估算，结果为经济损失值是湖区＞坝区＞泉山区，分别达到53786万元、44276.22万元、39247.36万元。坝区以林木死亡所造成的经济损失最大，湖区与泉山区以土地盐渍化所造成的经济损失最大。经济损失值占同期当地国民生产总值的百分比是湖区＞泉山区＞坝区，分别达到76.39％、55.04％、28.48％。可见绿洲的社会经济发展是以牺牲生态环境，降低生态安全水平为代价，这将严重制约绿洲今后的可持续发展。

Detection of Copper Ion with Laser-Induced Fluorescence in A Capillary Electrophoresis Microchip

A capillary electrophoresis microchip coupled with a confocal laser-induced fluorescence (LIF) detector was successfully constructed for the analysis of trace amounts of heavy metals in environmental sources. A new fluorescence dye, RBPhOH, synthesized from rhodamine B, was utilized in a glass microchip to selectively determine copper with high sensitivity. A series of factors including running buffer concentration, detection voltage, and sample loading time were optimized for maximum LIF detector response and, hence, method sensitivity.

Ultrafast dynamics of dye molecules in solution as a function of temperature

Femtosecond time-resolved studies using fluorescence depletion spectroscopy were performed on Rhodamine 700 in acetone solution and on Oxazine 750 in acetone and formamide solutions at different temperatures. The experimental curves that include both fast and slow processes have been fitted using a biexponential function. Time constants of the fast process, which corresponds to the intramolecular vibrational redistribution (IVR) of solute molecules, range from 300 to 420 fs and increase linearly as the temperature of the environment decreases. The difference of the average vibrational energy of solute molecules in the ground state at different temperatures is a possible reason that induces this IVR time-constant temperature dependence. However, the time constants of the slow process, which corresponds to the energy transfer from vibrational hot solute molecules to the surroundings occurred on a time scale of 1-50 ps, changed dramatically at lower temperature, nonlinearly increasing with the decrease of temperature. Because of the C-H...O hydrogen-bond between acetone molecules, it is more reasonable that acetone molecules start to be associated, which can influence the energy transfer between dye molecules and acetone molecules efficiently, even at temperatures far over the freezing point.

Ultrafast vibrational and thermal relaxation of dye molecules in solutions

Intra- and intermolecular relaxations of dye molecules are studied after the excitation to the high-lying excited states by a femtosecond laser pulse, using femtosecond time-resolved stimulated emission pumping fluorescence depletion spectroscopy (FS TR SEP FD). The biexponential decays indicate a rapid intramolecular vibrational redistribution (IVR) depopulation followed by a slower process, which was contributed by the energy transfer to the solvents and the solvation of the excited solutes. The time constants of IVR in both oxazine 750 and rhodamine 700 are at the 290-360 fs range, which are insensitive to the characters of solvents. The solvation of the excited solutes and the cooling of the hot solute molecules by collisional energy transfer to the surrounding takes place in the several picoseconds that strongly depend on the properties of solvents. The difference of Lewis basicity and states density of solvents is a possible reason to explain this solvent dependence. The more basic the solvent is, which means the more interaction between the solute and the neighboring solvent shell, the more rapid the intermolecular vibrational excess energy transfer from the solute to the surroundings and the solvation of the solutes are. The higher the states density of the solvent is, the more favorable the energy transfer between the solute and solvent molecules is.

施肥对黑土土壤微生物生物量碳的作用研究

长期施肥对黑土耕地表层土壤微生物生物量碳 (MBC)的作用研究结果表明 ,NPK化肥配施可保持休闲地土壤微生物生物量碳含量至 1.6g/kg水平 ;高量有机肥与无机肥配施可比休闲地土壤微生物生物量碳含量提高 1.96～ 2 .75倍 ;长期耕种与施肥对土壤微生物生物量碳含量产生衰减影响 ;各处理土壤微生物生物量碳含量增加依次为M2 +NPK(+14 1.2 5 % ,1990年始处理 ) >M4+NPK(+12 6 .88% ) >M2 +NPK(+10 1.2 5 % ,1980年始处理 ) >M4+CK(+80 .6 3% ) >(M1+NPK)× 1.5 (+13.13% ) >NPK(+8.12 % ) ,各处理土壤微生物生物量碳含量减少依次为M0+NPK(- 3.75 % ) >M1+NPK(- 17.5 0 % ) >M2 +CK(- 30 .6 3% ) >CK(- 4 7.5 0 % ) >M0 +CK(- 6 1.88% )。

糖蜜的可发酵性与制氢条件

为了提高糖蜜的可发酵性和氢转化率,采用批式发酵法研究了糖蜜前处理过程和发酵条件对Ethanoligenens sp B49生长和产氢的影响。结果表明去除煮沸驱氧过程可以大大提高Ethanoligenens sp B49发酵糖蜜产氢的性能和糖蜜的氢转化率。Ethanoligenens sp B49细胞生长和产氢的最佳COD为20.6g/L,可发酵糖蜜的最大耐受COD为41.2 g/L,酵母粉可以大大提高糖蜜的生物可利用性和比产氢率。COD为20.6g/L,外加酵母粉浓度为4g/L是Ethanoligenens sp B49发酵糖蜜产氢的最佳条件,单位体积产氢量达到78.97mmolH2/L培养基,比对照提高了76.2%。本研究改进糖蜜发酵前处理方法和发酵条件,可以大大提高了糖蜜制氢的生物可发酵性和比产氢率。

Sensitive turn-on fluorescent detection of cyanide based on the dissolution of fluorophore functionalized gold nanoparticles

We report a simple fluorescent method for sensitive cyanide detection based on the dissolution of Rhodamine B-adsorbed gold nanoparticles by cyanide.

Surface modification of poly(dimethylsiloxane) microchips using a double-chained cationic surfactant for efficiently resolving fluorescent dye adsorption

This paper described a double-chained cationic surfactant, didodecyldimethylammonium bromide (DDAB). for dynamic surface modification of poly(dimethylsiloxane) (PDMS) microchips to reduce the fluorescent dyes adsorption onto the microchannel. When DDAB with a high concentration was present as the dynamic modification reagent in the running and sample buffer, it not only reversed the direction of electroosmotic flow, but also efficiently suppressed fluorescent dyes pyronine Y (PY) or rhodamine 8 (RB) adsorption onto the chip surface. In addition, vesicles formed by DDAB in the buffer with higher surface charge density and electrophoretic mobility could provide wider migration window and potential for the separation of compounds with similar hydrophobicity. Factors affecting modification, such as pH and concentrations of the buffer, DDAB concentration in the buffer were investigated. Compared with commonly used single-chained cetyltrimethylammonium bromide, DDAB provided a better modification performance.

Synthesis and characterization of the titanium complexes bearing two regioisomeric trifluoromethyl-containing enaminoketonato ligands and their behavior in ethylene polymerization

A series of new titanium complexes bearing two regioisomeric trifluoromethyl-containing enaminoketonato ligands (3a-h and 6a-h), [PhN=CRCHC(CF3)O](2)TiCl2 (3a, R = Me; 3b, R = n-C5H11; 3c, R = i-Pr; 3d, R = Cy; 3e, R = t-Bu; 3f, R = CH=CHPh; 3g, R = Et; 3h, R = n-C11H23) and [PhN=C(CF3)CHC(R)O](2)TiCl2 (6a, R = Ph; 6b, R = n-C5H11; 6c, R = i-Pr; 6d, R = Cy; 6e, R = t-Bu; 6f, R = CH=CHPh; 6g, R = CHPh2; 6h, R = CF3) have been synthesized and characterized. X-ray crystal structures analyses suggest that complexes 3c-e and 6c-d all adopt a distorted octahedral geometry around the titanium center. Complexes 3c, 3d and 6c display a cis-configuration of the two chlorine atoms around the titanium center, while complex 6d shows a trans-configuration of the two chlorine atoms. Especially, the configurational isomers (cis and trans) of complex 3e were identified both in solution and in the solid state by NMR and X-ray analyses. With modified methylaluminoxane as a cocatalyst, all the complexes are active towards ethylene polymerization, and produce high molecular weight polymers.

Direct formation of giant vesicles from synthetic polypeptides

This report describes direct formation of giant vesicles from a series of poly(L-lysine)-block-poly(L-phenylalanine) (PLL-b-PPA) block copolymers from their water solution. These polymers are prepared by successive ring-opening polymerization (ROP) of the two alpha-amino acid N-carboxyanhydrides and then removing the side chain protecting groups by acidolysis. The structures of the copolymers are confirmed by nuclear magnetic resonance (NMR), differential scanning calorimetry (DSC), and size exclusion chromatography ( SEC). The vesicles are studied by atomic force microscopy (AFM), field emission scanning electron microscopy (ESEM), and confocal laser scanning microscopy (CLSM). Rhodamine B is used as a fluorescent probe to confirm the existence of the vesicle with an aqueous interior. The vesicle size is in the range 0.55-6 mu m, depending on the absolute and relative lengths of the two blocks, on initial polymer concentration, and on solution pH. The vesicles are still stable in water for 2 months after preparation. Addition of the copolymer to DNA solution results in complex formation with it. The complex assumes the morphology of irregular particles of less than 2 mu m. It is expected to be used in drug and gene delivery.

有机/无机纳米杂化材料负载钛催化剂及聚合研究 Ⅰ.乙烯聚合

以苯乙烯与马来酸酐无规共聚物 ( SMA)为有机组分 ,以正硅酸乙酯 ( TEOS)为无机组分 ,利用 sol- gel方法制得纳米杂化材料 ,以此为载体合成了杂化材料载体钛系催化剂 .利用 XPS、IR分析了载体与 Ti Cl4 的结合方式 ,并进行了乙烯聚合的研究 .结果表明 ,聚乙烯活性并不高( 1.72× 10 6g/h·mol)且分子量分布为 4 .0左右 ;堆密度范围是 0 .30～ 0 .39g/cm3.

烯丙基取代的铁系“茂后”烯烃聚合催化剂的合成及其催化乙烯聚合的研究

合成了烯丙基取代的含氮中性配体 [( p-C3H5) C12 H16]2 N3C9H9,然后与 Fe Cl2 反应合成含烯丙基取代的三齿“茂后”烯烃聚合催化剂 [( p-C3H5) ( C12 H16) ]2 N3C9H9Fe Cl2 ,通过 IR,1H NHR,EI-MS对化合物进行了表征 .研究了它催化乙烯聚合的能力 ,这种催化剂与 MAO组成的催化体系可在常压下催化乙烯聚合 ,活性最高达 1 .9× 1 0 6g PE/ ( mol Fe· h) .所得聚乙烯粘均分子量在 5× 1 0 4 ～ 2 .6× 1 0 5之间

Flow cytometry and ultrastructure of cryopreserved red seabream (Pagrus major) sperm

The objectives were to assess motility, fertilizing capacity, structural integrity, and mitochondrial function in fresh versus frozen-thawed (15% DMSO was used as a cryoprotectant) sperm from red seabrearn (Pagrus major). Mean (+/- S.D.) rates of motility, fertilization and hatching of frozen-thawed sperm were 81.0 +/- 5.4, 92.8 +/- 1.9, and 91.8 +/- 5.2%, respectively; for fresh sperm, they were 87.5 +/- 7.7, 95.8 +/- 2.4, and 93.8 +/- 4.2%. Although motility was lower in frozen-thawed versus fresh sperm (P < 0.05), there was no effect (P > 0.05) of cryopreservation on fertilization or hatching. Based on scanning and transmission electron microscopy, 77.8 +/- 5.6% of fresh sperm had normal morphology, whereas for frozen-thawed sperm, 63.0 +/- 7.2% had normal morphology, 20.6 +/- 3.1% were slightly damaged (e.g. swelling or rupture of head, mid-piece and tail region as well as mitochondria), and 16.4 +/- 4.2% were severely damaged. Sperm were stained with propidium iodide and Rhodamine 123 to assess plasma membrane integrity and mitochondrial function, respectively, and examined with flow cytometry. For fresh sperm, 83.9% had an intact membrane and functional mitochondria, whereas for frozen-thawed sperm, 74.8% had an intact membrane and functional mitochondria, 12.7% had a damaged membrane, 9.9% had nonfunctional mitochondria, and 2.6% had both a damaged membrane and nonfunctional mitochondria. In conclusion, ultrastructure and flow cytometry were valuable for assessment of frozen-thawed sperm quality; cryopreservation damaged the sperm but fertilizing ability was not significantly decreased. (c) 2007 Elsevier Inc. All rights reserved.

江河源区高山嵩草（Kobresia pygmaea）草甸植物和土壤碳、氮储量对覆被变化的响应

以青海省果洛州藏族自治州甘德县青珍乡高山嵩草Kobresia pygmaea草甸轻度退化草地和重度退化草地为研究对象,通过植物地上部分主要功能群（禾草类、杂类草、莎草类）、植物根系和土壤碳、氮浓度及储量动态研究,结果表明：高寒小嵩草草甸轻度退化草地地上部分主要功能群碳、氮浓度和C∶N比值明显高于重度退化草地的浓度。同一草地类型主要功能群比较,碳、氮浓度依次为杂类草〉禾草类〉莎草类;植物地上部分的碳、氮浓度明显高于地下根系的碳、氮浓度。重度退化草地植物根系碳、氮浓度高于轻度退化草地植物根系碳、氮浓度。重度退化草地土壤总有机碳浓度显著低于轻度退化草地土壤总有机碳浓度,随着土层的加深碳、氮浓度有减少的趋势。江河源区高山嵩草草甸的土壤有机碳、氮储量最大,植物根系碳、氮储量居中,植物地上部分碳、氮储量最小。重度退化草地总有机碳储量（13554.3g/m~2）较轻度退化草地储量（14669.2g/m~2）下降7.60%。其中,0-40cm土壤层碳储量下降4.10%,植物根系碳储量下降59.97%,植物地上部分碳储量下降15.39%;重度退化草地总氮储量（3780.6g/m~2）较轻度退化草地储量（3352.7g/m~2）高12.76%,其中,0-40cm土壤中总氮储量高13.07%,植物根系全氮储量下降55.09%,植物地上部分全氮下降16.00%。由于草地退化损失有机碳11149kg/hm~2,而全氮增加4278kg/hm~2。