云芝漆酶理化性质及动力学研究

主要研究pH值、金属离子、温度对云芝漆酶活性和稳定性的影响,以及该酶底物浓度效应和Km值测定.实验结果表明:Cu2+、Co2+对漆酶有激活作用,而Ag+则有抑制作用;该酶最适pH为4.8,在pH4~4.8表现出较强的稳定性;最适反应温度为40℃,低于50℃时有较好的热稳定性;Km值为4.2×10-3mol/L.     

改性离子交换树脂固定化漆酶及其动力学性质研究

环氧基改性的丙烯酸系树脂含环氧基,环氧基可在温和条件下与蛋白质上的亲核基团氨基反应而实现其固定化.研究了环氧基改性的离子交换树脂固定化漆酶的最优条件和固定化漆酶的酶学性质,结果表明,固定化漆酶时,在硫酸铵浓度为1.5mol·L-1、加酶量0.2U·g-1、pH 6.5、温度35℃条件下固定3h,测得固定化酶活性最好,比活力为0.026U·g-1,活力回收率为3.67%;该固定化酶催化反应的的最适温度为45℃,最适pH 4.5,米氏常数(Km)为1.233×10-3 mmol·L-1.锰离子、铁离子和钙离子在4×10-3~4×10-4 mol·L-1浓度之间时对固定化漆酶活性有抑制作用,在4×10-5~4×10-6 mol·L-1浓度之间时有激活作用.     

产普鲁兰酶克雷伯氏菌的分离鉴定及酶学性质研究

利用曲里苯蓝法从淀粉加工厂废水氧化池酸性污泥样品中筛选普鲁兰酶产生菌,对筛选到的GXAS-38菌株进行形态观察、生理生化特征分析、16SrDNA序列系统发育分析和普鲁兰酶酶学性质研究,并用PCR方法克隆GXAS-38菌株的普鲁兰酶基因。结果表明,GXAS-38菌株属于肺炎克雷伯氏菌(Klebsiella pneumoniae),其产普鲁兰酶的最适反应温度为60℃,在温度35～50℃时酶活较稳定;最适反应pH值5.5,在pH值5.0～7.5时酶活较稳定。Ca2+、Na+和Li+对GXAS-38菌株的普鲁兰酶活性有激活作用;Cu2+、Zn2+、Co2+、Mn2+、Fe2+、Fe3+和Ba2+对GXAS-38菌株的普鲁兰酶活性有抑制作用;螯合剂EDTA能够强烈地抑制GXAS-38菌株的普鲁兰酶活性,GXAS-38菌株的普鲁兰酶反应需要金属离子参与。GXAS-38菌株完整的普鲁兰酶编码基因全长3291bp,编码1096个氨基酸。     

荧光假单胞菌Pseudomonas fluorescens GcM5 1A菌株的 胞外木素过氧化物酶的部分纯化和鉴定

荧光假单胞菌GcM5 1A是松材线虫携带的一种致病菌。在GcM5 1A的培养液中发现有木质素过氧化物酶的活性，同时显示很强的毒性。通过对其胞外木质素过氧化物酶的酶学性质的初步研究发现：其最适温度为35 ℃，最适pH为4.0。在15~35 ℃、pH为6.0~9.0范围内，酶活保持相对稳定。NH2OH·HCl和KCN对酶活的抑制作用分别达到88 %和57 %，而NaN3对酶活的抑制作用只有15 %左右。Ca2+、Mn2+ 和Fe3+对其活性有增强作用，但是Hg2+ 和 Zn2+却有中等的抑制作用。KCN 和NH2OH·HCl抑制其酶活性分别达57 %、88 %，而NaN3的抑制率仅为15 %。吸收光谱显示该酶在406 nm处有一个Soret峰，加入1 mmol/L Na2S2O4后，吸光度出现了明显的下降，但Soret峰没有显著移动。     

重组PDOR基因的表达与纯化及部分酶学性质

将dhaT在E.coli BL21(DE3)pLysS中进行表达,并对含有His6标记的1,3-丙二醇氧化还原酶(PDOR)进行纯化.重组PDOR反应的最适pH值为10.0,最适温度为55℃,其以1,3-丙二醇和NAD+为底物的表观米氏常数Km值分别是15.5,0.23 mmol*L-1.实验表明,Ca2+, Mg2+和Cu2+对酶活性有抑制作用,而Fe2+, Na+, NH+4和Mn2+对酶活性有促进作用;1,3-丙二醇是PDOR适合的氧化底物.     

青霉葡萄糖氧化酶的分离纯化及性质研究

从青霉(Penicillium amagasakiense)发酵液中分离纯化葡萄糖氧化酶(简称GOD),通过DEAE-32离子交换柱层析、Sephacryl S-200分子筛凝胶过滤柱层析纯化,获得比活力为472 U/mg电泳单一纯酶制剂,采用SDS-PAGE电泳,表明该酶含有两个同型的亚基,分子量为150 ku.酶学性质研究表明:该酶催化葡萄糖氧化酶反应的最适pH为5.6,最适温度为40℃.酶的热稳定性研究表明:该酶在pH 5.5～8.5区间和温度低于50℃下稳定.金属离子对酶活力的影响表明:Na+、K+、Ca2+、Pb2+、Mn2+等对酶活力基本没有影响;Al3+、Zn2+对酶有一定的激活作用;Ag+、Hg2+、Mg2+、Cd2+、Cu2+、Co2+等对该酶活力有不同程度的抑制作用.该酶以葡萄糖为底物的米氏常数Km值为122.6 mmol/L,Vm为25.52 μmol/(L·min)(pH 7.0,37℃).     

牛小肠碱性磷酸酶部分性质研究

摘要：用正丁醇抽提、硫酸铵分级沉淀、EAE-32柱层析和Sephadex G-150 凝胶过滤,从牛小肠中分离纯化出牛小肠碱性磷酸酶（BIAP）.提纯倍数为50.69,比活力为48.87U/mg蛋白.酶液经SDS-PAGE呈现单一条带,且不含DNA核酸酶.该酶催化对硝基苯磷酸二钠（p-NPP）水解反应的最适pH值为9.7,pH小于6.5大于11.5均不稳定;最适温度为45℃,高于50℃不稳定.Km值为0.29mmol/L.45℃,pH9.7时最大反应速度（Vmax）为4.6μmol/(L•min).利用SDS-PAGE测定酶亚基的分子量为66KD. Mg2+、Mn2+和Ca2+对酶有不同程度的激活作用,Zn2+和EDTA对酶有抑制作用。随着Mg2+、Zn2+和EDTA浓度增加,270nm处紫外吸收值增加.     

芦荟叶皮β-葡萄糖苷酶的分离纯化及酶学性质

库拉索芦荟叶皮经匀浆、柠檬酸-柠檬酸钠缓冲液抽提,硫酸铵分级沉淀,DEAE-Sepharose fast flow层析和Superdex-200prep grade层析,获得电泳纯的β-葡萄糖苷酶(β-Glucosidase,BGL).纯化结果是β-葡萄糖苷酶比活力为98.48U/mg,纯化倍数为328.27倍,酶活性回收率为9.87%,全酶分子质量约为69.3kD,亚基分子质量约为69.7kD.酶学性质研究表明:β-葡萄糖苷酶的最适反应温度和最适反应pH值分别为40℃和5.0;在20~30℃及pH4.0~8.0范围内稳定性较好;最适条件下,以pNPG为底物的K_m值为2.21mmol/L,V_(max)为1.381μmol/(min·L).乙醇,抗坏血酸,Mn~(2+),K~+对该酶活性具有激活作用;SDS,Cu~(2+)对该酶活性具有强烈抑制作用;异丙醇,EDTA,尿素,Mg~(2+),Li+对该酶活性影响较小;甲醇对该酶有双重作用.     

大豆幼苗根系抗氧化酶活性的耐酸、铝研究

以大豆(029-289,浙春2号)为实验材料,在水培条件下,用不同量的酸、铝(T1:0 mmol·L-1 Al+pH 3;T2:0 mmol·L-1 Al+pH 4;T3:0 mmol·L-1 Al+pH 5;T4:0.5 mmol·L-1 Al+pH 5;T5:1 mmol·L-1Al+pH 5;T6:1 mmol·L-1 Al+pH 3)处理24,,61,2和24 h后,测定大豆幼苗根系的可溶性蛋白质含量与超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)的活性.结果显示:随着酸、铝处理时间的增加,大豆根系的可溶性蛋白质含量及SOD,POD和CAT的活性均呈先上升后下降的变化趋势,在酸、铝处理6 h时达到最大值,24 h时达到最小值.在无铝影响下,随着pH的升高,可溶性蛋白质含量及SOD,POD和CAT的活性先升高后降低,在T2处理下达到最大值.在pH 5条件下,随着铝浓度的增加,SOD活性先增大后减小,在T4处理下出现最大值;POD和CAT的活性随铝浓度的增加逐渐增大.在T6处理下,大豆根系SOD,POD和CAT的活性达到各酸、铝处理的最小值,说明该处理对大豆根系的伤害最大.     

大肠杆菌谷氨酸脱羧酶的纯化及其性质的研究

大肠杆菌菌体经机械研磨,硫酸铵分级,sephadex G-100层析,再用聚丙烯酰胺垂直板凝胶电泳进行纯化,得到电泳均一的 L-谷氨酸脱羧酶纯品.酶作用的最适pH 为4.4,在 pH3.6～5.8稳定;最适温度50℃,50℃以下稳定;于60℃保温30min,酶活力保留65%.此酶作用于 L-谷氨酸的 K_m 值为1.39×10~(-2)mol/L,金属离子 Zn~(2+),Cu~(2+),Mg~(2+),Fe~(2+)分别不同程度的抑制此酶,EDTA 无抑制作用.该酶在280nm 处有最大光吸收,与蛋白质在280nm 处有最大光吸收值的普遍性质相一致.     

Markedness and its Interpretation in Language Use

Language markedness is a common phenomenon in languages, and is reflected from hearing, vision and sense, i.e. the variation in the three aspects such as phonology, morphology and semantics. This paper focuses on the interpretation of markedness in language use following the three perspectives, i.e. pragmatic interpretation, psychological interpretation and cognitive interpretation, with an aim to define the function of markedness.     

低渗透非均质砂岩油藏启动压力梯度研究

理论推导与室内实验相结合,建立了低渗透非均质砂岩油藏启动压力梯度确定方法。首先借助油藏流场与电场相似的原理,推导了非均质砂岩油藏启动压力梯度计算公式。其次基于稳定流实验方法,建立了非均质砂岩油藏启动压力梯度测试方法。结果表明:低渗透非均质砂岩油藏的启动压力梯度确定遵循两个等效原则。平面非均质油藏的启动压力梯度等于各级渗透率段的启动压力梯度关于长度的加权平均;纵向非均质油藏的启动压力梯度等于各渗透率层的启动压力梯度关于渗透率与渗流面积乘积的加权平均。研究成果可用于有效指导低渗透非均质砂岩油藏的合理井距确定,促进该类油藏的高效开发。     

On Tragic Consciousness In The Works By Ernest Hemingway

As an American modern novelist who were famous in the literary world, Hemingway was not a person who always followed the trend but a sharp observer. At the same time, he was a tragedy maestro, he paid great attention on existence, fate and end-result. The dramatis personae's tragedy of his works was an extreme limit by all means tragedy on the meaning of fearless challenge that failed. The beauty of tragedy was not produced on the destruction of life, but now this kind of value was in the impact activity. They performed for the reader about the tragedy on challenging for the limit and the death.     

Achieving Unusual States of Matter Under High Pressure

正The periodicity of the elements and the non-reactivity of the inner-shell electrons are two related principles of chemistry,rooted in the atomic shell structure.Within compounds,Group I elements,for example,invariably assume the+1 oxidation state,and their chemical properties differ completely from those of the p-block elements.These general rules govern our understanding of chemical structures and reactions.Using first principles calcula-     

Exchange-Correlation and Electronic Excitation Energies from Pairing Matrix Fluctuations

We have developed an adiabatic connection to formulate the ground-state exchange-correlation energy in terms of pairing matrix linear fluctuations.This formulation of the exchange-correlation energy opens a new channel for density functional approximations based on the many-body perturbation theory.We illustrate the potential of such approaches with an approximation based on the particle-particle Random Phase Approximation(pp-RPA).This re-     

Density Functional Theory for the Response of Periodic Systems to Electric Fields Based on the Vector Potential Approach

正The electronic and nuclear(structural/vibrational)response of 1D-3D nanoscale systems to electric fields gives rise to a host of optical,mechanical,spectral,etc.properties that are of high theoretical and applied interest.Due to the computational difficulty of treating such large systems it is convenient to model them as infinite and periodic(at least,in first approximation).The fundamental theoretical/computational problem in doing so is that     

On the Thermodynamic Limit for Responses to Static Electromagnetic Fields

For molecular systems,the quantum-mechanical treatment of their responses to static electromagnetic fields usually employs a scalar-potential treatment of the electric field and a vector-potential treatment of the magnetic field.Although the potential for each field separately is associated with the choice of an(unphysical)origin,the precise choice of the origin for the electrostatic field has little consequences for the results.This is different for the     

An affordable accurate evaluation of the binding conformations predicted by molecular docking studies in bio-system

Franck-Condon factors bridge the gap between theoretical modeling and experimental observations for molecular electronic spectroscopy and electron transfer.Under the displaced harmonic oscillator approximation,multidimensional Franck-Condon factors are decomposed into a product of many one-dimensional(1D)Franck-Condon(FC)factors,and each 1D-FC factor is associated with one Huang-Rhys factor that determines the leading contribution of     

Call for Papers

<正>"The Journal of Shanghai Normal University:Mathematics"is published by Shanghai Normal University as regular issues of The Journal of Shanghai Normal University each year from 2014 in English.The editors-in-chief of the issues are professors Yuhao Cong and Maoan Han.The Journal of Shanghai Normal University was started in 1958 with     

Journal of Donghua University ( English Edition) Contents of Volume 31

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