修饰脂肪酶催化水解反应的动力学和热力学研究

采用邻苯二甲酸酐(PA)对猪胰脂肪酶(PPL)进行了化学修饰,以三硝基苯磺酸法测定修饰酶的平均氨基修饰度,并比较了酶修饰前后的紫外光谱和荧光光谱;采用动力学方法考察了酸碱和热对修饰酶水解活性和稳定性的影响,并计算了一系列动力学和热力学参数.实验结果表明:PA对PPL的修饰度为30%,脂肪酶PPL经PA修饰后,其紫外特征吸收峰发生红移,吸收强度降低,而且最大荧光发射峰发生蓝移,荧光强度增大;PPL经PA修饰前后,催化水解反应的最适pH和最适温度未发生变化,仍为pH7.5和40℃,但修饰后酶催化水解的效率和酶与底物的亲和力得到有效提高,反应的活化能Ea、自由能ΔG≠、活化焓ΔH≠及活化熵ΔS≠均明显降低,如最适条件下修饰酶PA-PPL和脂肪酶PPL的转化数kcat和反应的活化能Ea分别为39.8 s-1、25.8 kJ/mol和34.4s-1、43.8kJ/mol;PPL经PA修饰后,酶分子热变性的热力学参数(变性活化能Ea,d、变性自由能ΔGd≠、变性活化焓ΔHd≠及变性活化熵ΔSd≠)均增大,半衰期t1/2延长,酶分子的热稳定性也得到有效提高.     

丁二酸酐修饰对漆酶稳定性和除酚效率的影响(英文)

采用(NH4)2SO4分步沉淀法对诺维信中国公司生产的漆酶制剂DeniLiteIIS进行了纯化,并用丁二酸酐(SA)对纯化酶进行了化学修饰,运用三硝基苯磺酸法、紫外光谱法及荧光光谱法对修饰效果进行了初步表征,比较了天然酶和修饰酶的pH稳定性、热稳定性及除酚效率.结果表明,修饰酶的平均氨基修饰度为85%,其紫外吸收峰和荧光发射峰均出现蓝移,而且紫外吸收减小、荧光强度增加.尽管采用SA化学修饰未能改变漆酶的最适反应温度,但使其最适反应pH值由4.5提高到5.5,并且使酶活提高60%.与天然酶相比,修饰酶的pH稳定性和热稳定性更高,催化效率(kcat)和酶与底物的亲和力(kcat/Km)分别提高了53%和122%,对邻、间和对苯二酚的除酚效率分别提高了48%,57%和18%.这预示着这些修饰漆酶可望应用于工业生产和酚类污染废水的治理.     

木瓜蛋白酶的化学修饰及对其酶活力的影响

用不同酸酐对木瓜蛋白酶进行化学修饰, 以三硝基苯磺酸法(TNBS)测定修饰酶的平均氨基修饰度, 对修饰前后的木瓜蛋白酶分别纯化并通过UV-vis和IR对其结构进行了表征. 考察了温度、pH值和表面活性剂SDS对化学修饰的木瓜蛋白酶活力的影响, 并与天然木瓜蛋白酶进行了比较, 对天然酶和修饰酶进行了动力学研究. 结果表明, 化学修饰木瓜蛋白酶的最适反应温度为80 ℃; 最适pH值为9.0; 在SDS浓度为5 mg•mL－1时修饰酶酶活仍能保持在50%左右; 在所有酶中, 均苯四甲酸酐修饰木瓜蛋白酶的催化效率最高, 为2.442×102. 与天然木瓜蛋白酶相比, 化学修饰木瓜蛋白酶的热稳定性、耐碱性和耐洗涤性得到了显著提高.     

甲壳素对铅(Ⅱ)的吸附研究

进行了铅离子在甲壳素上的吸附行为实验,结果表明在pH=4.46时吸附最佳。测得静态饱和吸附容量为478mg.g-1(树脂);用0.5mol.L-1HCl可洗脱,洗脱率达98.8%;测得表观速率常数k298=8.53×10-5s-1;表观活化能Ea=16.54kJ.mol-1;等温吸附服从Freundlich经验式;吸附热力学参数△H=8.70kJ.mol-1,△S=41.7J.mol-1.K-1,△G=-3.71kJ.mol-1。     

化学修饰木瓜蛋白酶的固定化及性质研究

在底物保护和无底物保护下,用丁二酸酐对木瓜蛋白酶进行化学修饰,以三硝基苯磺酸法测定修饰酶的平均氨基修饰度,以棉布为载体,戊二醛为交联剂,对修饰前后的木瓜蛋白酶分别进行固定化.考察了温度、pH和表面活性剂SDS对化学修饰的固定化木瓜蛋白酶活力的影响,并与固定化天然木瓜蛋白酶进行了比较.研究表明,化学修饰固定化木瓜蛋白酶的最适反应温度为80℃;最适pH为9.0;在SDS浓度为20mg/mL时酶活也仍能保持在40%左右;米氏常数为187g/L.与天然的固定化酶相比,化学修饰的固定化木瓜蛋白酶的热稳定性、耐碱性和耐洗涤性得到了显著提高.     

5-氟胞嘧啶气相及水助质子转移异构化的理论研究

采用密度泛函B3LYP/6-311G**方法,对6种5-氟胞嘧啶异构体孤立分子的稳定性及质子转移引起的酮式-烯醇式、氨基式-亚胺式互变异构反应机理进行了计算研究,获得了零点能、吉布斯自由能及质子转移过程的反应焓、活化能、活化吉布斯自由能和速率常数等参数.计算结果表明,气相中烯醇-氨基式FC4是最稳定的异构体.分子内质子转移设计了FC1→FC2和FC1→FC6两条通道,分别标记为P(1)和P(2),各通道速控步骤的活化能和速率常数分别为155.9 kJ·mol-1,4.70×10-15 s-1和173.1 kJ·mol-1,1.41×10-18 s-1.水助催化时,相应通道P(3) 和P(4) 速控步骤的活化能和速率常数分别为51.0 kJ·mol-1,1.41×103 s-1和88.2 kJ·mol-1,4.53×10-3 s-1.可见,水分子的加入极大地降低了质子转移的活化能垒.另外发现,水分子参与形成协同的双质子转移机理比水助单质子转移机理更利于降低活化能垒.     

氨基膦酸树脂对溶液中锌的吸附研究

研究了Zn2+在氨基膦酸树脂上的吸附行为。结果表明,静态饱和吸附容量为209.2mg.g-1树脂,用2.0 mol.L-1的HCl溶液洗脱,洗脱率为98%以上;测得吸附热力学参数分别为:ΔH=8.14 kJ.mol-1,ΔG=-3.06 kJ.mol-1,ΔS=37.6 J.mol-1.K-1。等温吸附服从Freundlich吸附等温式;表观活化能Ea=14.7 kJ.mol-1,表观速率常数k298=4.26×10-5s-1;树脂功能基与Zn2+的配位比约为1∶1;并用化学和红外光谱的方法探讨了树脂对Zn2+的吸附机理。     

邻苯二甲酸酐化学修饰对辣根过氧化物酶催化活性的影响

通过采用邻苯二甲酸酐(PA)对辣根过氧化物酶(HRP)的蛋白链进行修饰, 研究了PA化学修饰对HRP的稳定性、催化活性、活性中心结构、酶对底物的亲合性和专一性等催化性质的影响. 结果显示: 酸性条件下(pH=3), 4小时后PA-HRP的催化活性比天然HRP提高了7.5%;碱性条件下(pH=10), 4小时后PA-HRP的催化活性比天然HRP提高了27%. PA-HRP的K_m值为8.16 (mmol/L), 小于天然HRP的K_m值12.99 (mmol/L), 而PA-HRP的k_(cat)/K_m值为7.86(10~4(L/ mmol· min)大于天然HRP的k_(cat)/K_m的6.70(10~4(L/ mmol· min). 这些催化活性和动力学数据表明了PA-HRP与天然HRP相比其稳定性、酶对底物的亲和性和专一性得到了提高. 紫外-可见光谱、拉曼光谱数据显示: 修饰剂PA 改变了天然HRP血红素周围的微环境, 对酶蛋白分子的活性中心结构并没有影响. 差示光谱显示PA修饰剂可以提高酶对底物的亲和力.     

木瓜蛋白酶交联聚体的制备及性质

采用交联酶聚体(CLEAs)技术制得了木瓜蛋白酶CLEAs, 优化了制备条件. 以纯乙醇为蛋白沉淀剂, 质量分数40%戊二醛为交联剂, 于4 ℃下对酶沉淀聚体交联16 h; 所得木瓜蛋白酶CLEAs的最适pH为6.0(游离酶最适pH=7.0), 最适温度范围由游离酶的80 ℃拓宽为50～80 ℃, 热稳定性和溶液稳定性亦明显提高; 微观形貌分析证明木瓜蛋白酶CLEAs优良的催化效能及稳定性来自于CLEAs单元所具有的高比表面积及单元内部多点共价固定的结合方式.     

毛细管电泳法应用于阿魏酸异构化动力学和热力学研究

基于毛细管电泳技术,建立了阿魏酸异构化反应的动力学和热力学研究方法.测定了阿魏酸不同温度下异构化反应速率常数;基于阿仑尼乌斯经验公式和埃林方程,分别计算出阿魏酸异构化反应的活化能(阿仑尼乌斯经验公式Ea1:43.38kJ·mol-1,Ea-1:58.51kJ·mol-1;埃林方程Ea1:43.24kJ·mol-1,Ea-1:58.36kJ·mol-1;从理论上证明了在常温下阿魏酸异构化是一个自发的过程,其吉布斯自由能变化△(△G≠298.15K)=-6.12kJ·mol-1.     

Fluorescent modification for peptide sequencing by postsource decay-matrix assisted laser desorption/ionization-mass spectrometry

The sequential analysis of a peptide of CDYEGRLI, relating to the nucleic proteins in influenza virus, was performed by the postsource decay (PSD) fragmentation method using matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). The sequence of the peptide was difficult to analyze by MALDI-MS since the PSD fragment ions of the peptide were almost never observed and were not amenable to complete sequence interpretation. The peptide was modified by 4(5)-(iodoacetamide) fluorescent reagent to improve the sensitivity of the MALDI-PSD fragment spectrum. In the spectrum of the fluorescent modified peptide, almost all sequential b-series fragment ions were observed clearly, which was sufficient for complete sequence interpretation. The results indicate the advantage of fluorescent modification for the total sequencing of the peptides by MALDI-MS.     

模板法制备定向排列的聚乙烯吡咯烷酮纳米管簇

近年来,随着纳米科学的迅猛发展,对定向生长的纳米碳管、半导体、氧化物及金属纳米线、管等无机材料的制备引起人们广泛的关注．然而对定向生长的聚合物纳米结构材料如聚合物的纳米管、线等的制备,却未见报道．最近,德国马普微结构研究所通过模板法制得了具有取向一致的聚合物纳米结构材料,该材料在化学传感器、药物输送以及微环境研究等方面具有广泛的应用前景．     

Adsorption of Bismarck Brown by Iron Oxide Nanosphere and Its Modified Form

In the present study, adsorption of Bismarck Brown (BB) dye onto iron oxide nanospheres (IONs) and modified IONs by HCl from aqueous solution was investigated. The IONs was synthesized by solvothermal method and then modified by HCl. The high magnetic properties of both adsorbents lead to facial separation from aqueous solution by an external magnet. The results show that the modification of adsorbent cause higher adsorption capacity for removal of BB from aqueous solution. The prepared adsorbents were characterized by SEM, XRD, FTIR, and ATR-IR techniques. The adsorption kinetic and equilibrium data were fitted with different models. The results show that the equilibrium and kinetic data were best fitted with Langmuir–Freundlich isotherm and fractal-like pseudo-second-order kinetic model, respectively. The effects of pH and temperature have also been investigated.     

Sorption kinetic studies of U(VI) on inorganic and organic modified red earth: comparison of linear and non-linear methods

The sorption of U(VI) on natural red earth, inorganic modified red earth and organic modified red earth were studied under given conditions. Kinetic studies of U(VI) sorption onto the red earth samples were discussed by linear and non-linear approaches. The results indicated that the inorganic modification greatly improved the surface properties and the sorption kinetics characteristics, and that organic modification increased the sorption capacity significantly. The non-linear pseudo-second-order kinetic model could fit the kinetics much better compared with the other linear forms.

     

十八烷基三氯硅烷表面改性钛酸钾晶须

利用十八烷基三氯硅烷(OTS)的自组装对钛酸钾晶须(PTW)进行表面改性. 通过测定苯、乙二醇、水在OTS改性的PTW压片上的接触角, 计算了OTS改性PTW后的表面能和极性分量, 并与传统方法(PMMA包裹和KH560表面改性)改性的PTW进行比较, 结果表明, OTS改性的PTW具有更低的表面能和极性分量, 以及更好的亲油疏水性能. IR分析和色散分量测定表明, 改性后OTS的烷基链倾斜在PTW表面.     

A novel Surface Plasmon Resonance enhanced Total Internal Reflection Ellipsometric application: electrochemically grafted isophthalic acid nanofilm on gold surface

The scope of this study is to modify a Surface Plasmon Resonance (SPR) sensor slide with isophthalic acid to evaluate the possible application on the detection of copper(II) ions in aqueous media by total internal reflection ellipsometry. A gold sensor surface was modified by an electrochemical diazonium reduction modification method. The modified surfaces are characterized with cyclic voltammetry (CV) and ellipsometry. Isophthalic acid monolayer modified gold slides were used for in situ detection of aqueous Cu(2+) solution with the SPR enhanced total internal reflection ellipsometry (SPRe-TIRE) technique. Layer formation, pH dependency of adsorption, sensor response of the SPRe-TIRE and isothermal kinetic parameters were examined. A high dependency on the number of CV cycles in the monolayer-multiple layer transition was observed. The suggested sensor gave a linear response over a wide range of Cu(2+) concentrations. It was also reported that adsorption on the SPRe-TIRE sensor gave Langmuir adsorption model behavior.     

A new action photoelectron spectroscopy for anions

We present a new experimental approach, in which anion photodetachment spectroscopy is recorded with electrons of fixed kinetic energy. This approach circumvents some shortcomings of the zero electron kinetic energy method. Our method is based on a modified magnetic bottle photoelectron spectrometer (MBPES). A tunable laser is used to detach electrons from mass selected anions, drifting collinearly with the 40 cm MBPES drift tube. To avoid Doppler broadening, a low voltage pulse removes the velocity component of anions from the detached electrons. Spectra are recorded by collecting the wavelength dependence of electron-signal at a predetermined TOF window, corresponding to a specific electron-kinetic energy. We call this approach PEACE, denoting photoelectron action spectroscopy at constant kinetic energy. Our best resolution is 0.65 meV for 1.5 meV electrons. We present a PEACE spectrum of HgCl(-) together with the corresponding simulated theoretical spectrum. The method is similar in resolution and data collection rates to the slow electron velocity map imaging technique recently introduced by Neumark and co-workers.     

Grafting of Acrylamide and Trimethylolpropane Triglycidyl Ether onto the Gellan Gum: Synthesis and Characterization

To improve the physiochemical properties of gellan gum(GG), GG was modified with acrylamide and trimethylolpropane triglycidyl ether(TTE). The structure and morphology of modified GG were characterized by Fourier transform infrared(FT-IR) spectroscopy, X-ray diffraction(XRD), differential scanning calorimetry(DSC) and scanning electron microscopy(SEM). The characteristic peaks at 3448, 2788, 1654, 1411, 1117 and 1044 cm~(-1) in the FT-IR spectrum confirm the modification. The XRD and DSC data revealed that the modification enhanced the thermal stability of GG. SEM analysis suggested the modification introduced a porous microstructure, resulting in the adsorption of crystal violet. In addition, the adsorption capacity, thermal stability and swelling property of GGTTE3 were superior to GGTTE1, GGTTE2, GGTTE4 and GGTTE5.     

改性埃洛石材料的制备及其对亚甲基蓝吸附行为的研究

使用硅烷偶联剂KH550改性埃洛石纳米管获得改性材料HNTs-APTS,并对其吸附亚甲基蓝的行为进行研究。利用傅立叶变换红外光谱仪(FTIR)、X-衍射仪(XRD)对改性前后的埃洛石进行表征。考察了吸附时间和温度对吸附过程的影响,并采用Lagrange准二级动力学方程、Langmuir等温线方程及Freundlich等温线方程对实验数据进行拟合。结果表明,KH550成功负载到埃洛石表面;改性后材料的吸附能力大大提高。改性埃洛石对亚甲基蓝的吸附约在60 min达平衡,最大吸附容量为21.66 mg/g。其吸附过程符合准二级动力学方程,热力学较好地符合Langmuir等温线方程,且吸附过程为自发吸热,升高温度有利于吸附的进行。改性材料可重复再生6次,具有良好的再生性能,可在工业处理亚甲基蓝废水中使用。     

表面活性剂改性有机沸石对废水中亚甲基蓝的吸附行为

用表面活性剂十六烷基三甲基溴化铵(CTMAB)和十二烷基硫酸钠(SDS)复合改性沸石,制备了阴阳离子表面活性剂改性沸石,利用X射线衍射和红外吸收光谱表征了改性前后沸石的结构,并研究了改性沸石对亚甲基蓝的吸附行为.结果表明,改性后沸石的吸附性能明显增强;在溶液pH 8、常温,吸附时间100 min、改性沸石用量20 g/...