疏绵状嗜热丝孢菌糖化酶基因（gla）的克隆及在毕赤酵母中的表达

本研究以疏绵状嗜热丝孢菌(Thermomyces laltltginosus)cDNA为模板,克隆了糖化酶基因(gla),序列分析表明gla的开放阅读框由1854个核苷酸组成,编码617个氨基酸.根据氨基酸序列推算该酶的分子量为64 kD,属于糖苷水解酶第15家族,具有该家族催化保守区的典型特征.PCR扩增gla的成熟蛋白编码基因,构建表达载体,经线性化后电击转化导入巴斯德毕赤酵母(Pichia pastoris GS115),并成功进行了表达.重组酶经摇瓶发酵后酶活可达11.6 U/mL,经硫酸铵沉淀、DEAE-Sepharose Fast Flow阴离子交换等步骤纯化了该重组蛋白,SDS-PAGE显示该重组蛋白大小约为67kD,比推测的蛋白分子量稍大,可能与该蛋白的糖基化有关.该重组酶的最适反应温度和最适pH值分别为60℃和5.0,该酶具有较高的热稳定性,70℃保温20 min后剩余酶活为54%.     

玉米秸秆热解动力学分析(简报)

该文采用耐驰STA449同步热分析仪,研究了玉米秸秆在升温速率为5、10、15、20、30 K/min,反应终止温度1273K的热分解反应,结果表明玉米秸秆热解过程可分为失水预热解、热解和碳化3个阶段,随着升温速率增加,反应的特征温度和最大失重速率增加,差示扫描（DSC）曲线整体向下倾斜,升温速率过大时,出现失水滞后现象。分别用FWO法、FRL法和Kissinger法对玉米秸秆热解进行了动力学计算,其热解活化能为（161±23）kJ/mol,通过Malek法确定了玉米秸秆热解满足J-M-A方程,反应机理为随机成核随后生长,确定了反应级数n和指前因子对数lgA的范围,并利用Matlab软件对实验数据进行拟合,验证了机理的正确性,并确定了玉米秸秆热解反应的动力学参数。该文结果对工程应用有指导意义。     

向日葵籽壳热解反应动力学的研究

采用热重分析法(TG)在不同升温速率下(5、10、20、30℃/min)对粒径为0.154～0.280 mm向日葵籽壳热解的热失重行为进行了研究。结果表明,向日葵籽壳热解分为四个阶段,随着升温速率的提高,各个阶段的起始和终止温度向高温侧稍微移动,并且主反应区间也略有增加。Ozawa法和Starink法计算得出的向日葵籽壳在热解过程中不同失重率下的活化能(E)都集中在140～178 kJ/mol范围内。用积分法Coats-Redfern方程、微分法Achar方程以及热分析动力学三因子求算的比较法得出该反应过程的机理函数表达式。经过对41种常用机理函数一一代入得出Jander方程能较好地描述向日葵籽壳热解反应过程,机理为三维扩散,球形对称,反应级数n=2。该研究可为生物质热解装置的工艺参数优化提供参考。     

城市土壤重金属有机酸淋溶特征与动力学机制

以典型工业城陕西省铜川市土壤为研究对象,采用BCR(欧共体标准物质局)连续形态分级法和Hakanson潜在风险指数法分别研究了人类活动性元素Pb,Cu,Cr,Zn的形态分布特征、环境迁移与生态风险;利用振荡浸提动力学试验(25℃平衡振荡24h)探讨了人工合成低氨基多羧酸(APCAs)乙二胺四乙酸(EDTA)、二乙烯三胺五乙酸(DTPA)和低分子有机酸(LMWOAs)抗坏血酸(Vc)对重金属Pb,Cu,Cr,Zn的淋溶解吸特征与动力学机制。结果表明,铜川市工业区土壤重金属污染程度严重,总体潜在生态风险程度较强;重金属Zn和Cu的环境迁移趋势较大。淋溶动力学试验表明,不同类型有机酸对土壤Pb,Cr,Cu和Zn的淋溶解吸具有相似的动力学过程,但淋溶速率和解吸量随有机酸的不同各异。重金属Pb,Cr,Cu和Zn各形态量之和与有机酸EDTA和DTPA淋溶解吸量呈指数关系,而与低分子有机酸Vc淋溶解吸量成线性关系。从而进一步表明低分子有机酸Vc与氨基多羧酸类有机酸EDTA和DTPA对土壤重金属Pb,Cr,Cu,Zn淋溶解吸机制不同。     

升温速率对椰壳热解特性的影响及动力学分析

利用热重分析仪对椰壳在不同升温速率（5、20、40℃/min）下的热解特性及动力学进行了研究,探讨了热解机理。结果表明：椰壳热解过程主要分为脱水、快速热解和缓慢失重;对比不同升温速率下的失重曲线表明,升温速率对热解失重率有明显影响,其最终热解产物的得率随升温速率的增加而减少。运用氮吸附仪测定其吸附等温线,获得不同热解温度对炭化料的孔结构,结果表明：热解温度越高,微孔越发达。使用TG曲线数据,利用分布活化能模型求出相应的活化能,失重率在0.1～0.8之间时,活化能在146～444 kJ/mol,呈“N”形变化。活化能的分布函数,反映了椰壳热解过程中不同阶段反应性能的变化规律,有助于了解椰壳的热解机理。     

氯离子在土壤中的扩散: Ⅱ.动力学及热力学性质

The amounts of chloride ions diffused in four soils of different textures at the same water content under different temperature and at varied time were measured by the diffusion cell method using ³⁶Cl-labelled CaCl₂ solution. Five kinetic models were used to fit the dynamic process of the diffusion of chloride ions in the soils. It was found that Elovich equation or power function equation was the best model to describe the process. The pseudothermodynamic parameters, i.e. the net reaction energy, the activation entropy, activation enthalpy and activation free energy of the diffusion, were derived from the absolute reaction-rate theory. The results showed that these parameters decreased in the order of loessal soil > black lu soil > lou soil > yellow cinnamon soil, which indicated that the force and the heat-energy barrier to be overcome for diffusion decreased, the diffusion rate increased and the disorder of the soil-solution-ion system due to diffusion decreased successively with the texture becoming heavier in the four soils.     

Kinetic properties and stability of potato acid phosphatase immobilized on Ca-polygalacturonate

 Acid phosphatase from potato was adsorbed and immobilized on a pre-formed network of Ca-polygalacturonate, a substrate which has a composition and morphology similar to the mucigel present at the root-soil interface. The influence of different types of organic buffers on enzyme adsorption and activity was investigated. The highest enzyme activity, for free and adsorbed enzyme, was obtained with Na-maleate buffer at pH 6.0, which was used for all subsequent experiments. The Michaelis-Menten kinetic parameters, V_max and K_m, were determined for free and adsorbed phosphatase. V_max showed a 60% decrease upon adsorption (2.09±0.30 U/mg, for the soluble form and 0.84±0.15 U/mg, for the adsorbed enzyme), whereas K_m increased from 0.49±0.15 mM for the free enzyme to 0.99±0.20 mM for adsorbed phosphatase. Phosphatase adsorption decreased as the concentration of NaCl increased, indicating that the enzyme is bound to the carrier gel through coulombic interactions. Adsorption increased stability of the enzyme as compared with the free enzyme (t _1/2 of the activity was 9.4 days and 5.8 days, respectively), but increased thermal and proteolytic inactivation. The pH/activity profile revealed no change in terms of shape or optimum pH (4.5) upon adsorption of the enzyme. These results indicate that adsorption of acid phosphatase on Ca-polygalacturonate induces changes in the kinetic properties and stability of the enzyme, and the same type of response can be extrapolated from these results for acid phosphatases of the rhizosphere. Received: 1 July 1997     

空心菜对不同形态氮吸收动力学特性研究

对空心菜不同形态氮素吸收动力学特性进行了研究,发现其吸附曲线均符合Michaelis-Menten酶动力学方程,采用LB转换式处理数据对其动力学参数进行计算,得出以下结论:①空心菜对铵态氮的亲和力大于对硝态氮的亲和力（K_m铵态氮<K_m硝态氮）,说明空心菜有优先吸收铵态氮的趋势,表明在实际净化水质过程中,若有足够的停留时间,空心菜对铵态氮的净化程度更高一些;②空心菜对铵态氮的最大吸收速率V_max为0.001 85 mmol/（g·FW·h）,对硝态氮最大吸收速率V_max 为0.000 279 mmol/（g·FW·h）,V_max铵态氮>V_max硝态氮,表明空心菜对铵态氮的净化速率要大于对硝态氮的净化速率;③当溶液中存在铵态氮时,空心菜对硝态氮的亲和力增加了2.5%,而最大吸收速率则减少34%,总的吸收能力降低,表明溶液中铵态氮的存在不影响空心菜根细胞膜上载体与硝态氮之间的亲和性,但影响载体对硝态氮的运转速度。     

Kinetic parameters of dehydrogenase in the assessment of the response of soil to vermicompost and inorganic fertilisers

 Kinetic parameters (V _max and K _m) of dehydrogenase activity were determined in order to assess the metabolic response of a soil about 1 year after organic and mineral treatments. The soil was planted with maize (Zea mays) and treated with the following fertilisers: organic (vermicompost; VC), mineral (ammonium phosphate and urea), and an organo-mineral mixture. V _max, which represents a measurement of the quantity of enzyme, markedly increased in organic and organo-mineral treatments, indicating that the addition of organic matter caused an increase in dehydrogenase in the active microbial biomass. K _m, representing enzyme-substrate affinity and/or different sources of the enzymes, was similar in VC-treated soil and control soil, while it doubled in organo-mineral and mineral treatments. These results suggest that the use of VC did not alter the enzyme-substrate affinity, while mineral fertiliser reduced this affinity or changed the composition and activity of soil microbiota. A positive correlation was found between V _max, the metabolic index (dehydrogenase/water-soluble carbon ratio), and the soil organic matter content. The kinetic constants of dehydrogenase activity and the metabolic index may be considered valid parameters to monitor the evolution of microbiological activity in soil. Received: 4 February 2000     

亚热带稻田土壤反硝化动力学参数估算

反硝化作用是土壤氮素损失的重要途径,对反硝化潜势的准确估算是农业精准施肥的必然要求。以亚热带典型红壤稻田土作为研究对象,足量添加外源氮进行室内淹水厌氧培养获取反硝化作用动态,并分别用米氏方程和一级动力学方程对其拟合,最后利用土壤基本理化性质对反硝化动力学参数进行估算。结果表明:米氏方程更适合反硝化动力学拟合,最佳的米氏常数(Km)为35mg/kg;米氏最大速率常数(vmax)与一级动力学速率常数(K)具有显著的相关性(r=0.96, P0.05)。土壤总氮,砂粒和粉粒以及土壤容重对vmax影响最大。利用总氮和粉粒含量作为输入参数估算了vmax,准确度达66%。所构建的参数方程既充分挖掘了土壤基础数据潜能,又能快速地获取土壤反硝化动力学曲线,省时省力。