Kinetic of Adsorption of Urea Nitrogen onto Chitosan Coated Dialdehyde Cellulose under Catalysis of Immobilized Urease

The adsorption of urea nitrogen onto chitosan coated dialdehyde cellulose (CDAC) under catalysis of immobilized urease in gelatin membrane (IE) was studied in batch system. The pseudo first-order and second-order kinetic models were used to describe the kinetic data, and the rate constants were evaluated. The experimental data fitted well to the second-order kinetic model.     

尿素酶固载于纳米二氧化钛多孔膜上的尿素生物传感器

在钛丝基体上沉积一层纳米二氧化钛(TiO2)多孔膜,然后直接将尿素酶吸附在TiO2膜上。基于TiO2膜的pH响应,发展了一种廉价的、易于微型化的pH敏尿素酶传感器。采用石英微天平、红外光谱和紫外可见光谱等手段研究尿素酶在TiO2膜上的物理固载行为。由于纳米TiO2在紫外光下光自洁特性可获得高度洁净的表面,石英微天平研究表明在光自洁后的TiO2膜上尿素酶的吸附具有很好的重复性和稳定性,吸附量为0.22mmol/g,吸附平衡常数k为3.15×105L/mol。采用电位法测定了尿素酶/TiO2复合膜电极的性能及其影响因素,在1.0 mmol/L pH 7~8 PBS,35℃,尿素的响应范围为8.5×10-5~1.5×10-1mol/L,相关系数r为0.993 7,检出限为6×10-5mol/L。     

Preparation of a Cyclomaltoheptaose(β-cyclodextrin) Cross-linked Chitosan Derivative via Glyoxal or Glutaraldehyde

Chitosan has been regarded as a multifunctional polymer applicable to various purpos-es1. The potential of the material in medical applications, particularly in drug-delivery formu-lations has been demonstrated2, 3. Recently, a crosslinked chitosan with various crosslink-ing agents was prepared as adsorbents4-7. A -cyclodextrin (?CD) has been subjected to numerous studies, particular in terms of its complexing ability with a variety of components8, 9. The complexing ability of -CD has b…     

以甲壳胺为载体的固定化嗜热菌蛋白酶的研究

本文报道了以甲壳胺为载体,采用戊二醛交联的方法将嗜热菌蛋白酶进行固定化。以酪蛋白为废物。固定化酶的活力回收为38％。本工作研究了各种因素对固定化酶活力的影响,实验结果表明:固定化后的酶,在热稳定性、抗抑制作用等方面较原酶有一定的提高。     

壳聚糖水凝胶微球的制备与溶胀性能

采用三聚磷酸钠(TPP)和六偏磷酸钠(SHMP)为复合交联剂,制备了离子交联网络结构的壳聚糖水凝胶微球。分析讨论了交联剂质量配比、pH以及离子强度等对壳聚糖微球溶胀性能的影响。结果表明:使用复合交联剂制备的微球的溶胀度比单独使用TPP或SHMP的分别低62.4%和41.3%,交联效率得到明显提高;当m(TPP)∶m(SHMP)=3∶5,交联剂pH=5时,制备的微球交联程度最好,结构最密实;在pH=1.2的缓冲溶液中,溶胀度可达到357%,而且凝胶没有任何破碎。微球具有较好的离子强度和pH敏感性,在酸性介质中有较高的溶胀度。     

京尼平交联磁性壳聚糖微球的制备及其脂肪酶的固定化

采用反相悬浮法与溶胶凝胶法结合制备磁性壳聚糖微球,并以此为载体,京尼平为交联剂,脂肪酶为模型酶进行固定化,研究了酶固定化的最优条件和固定化酶的性质。结果表明,在京尼平浓度为0.6 g/L、交联温度为55 ℃、交联时间8 h,固定化酶的比活力最大,为4.31 U/g。固定化酶在25~35 ℃,pH值在8.0有最大活性,其米氏常数Km为0.26 mol/L。同时,固定化酶具有良好的热稳定性及pH稳定性,可重复利用,且能进行磁分离。     

交联血管紧张素转化酶聚集体的制备及性质

制备了交联血管紧张素转化酶聚集体（ACE-CLEAs）,比较了ACE-CLEAs及游离ACE的酶学性质,包括最适酶促反应温度、最适pH值、Km、vmax、温度稳定性及pH稳定性等。 以酶活力回收率为参考,确定了制备ACE-CLEAs的最佳条件为：饱和度为80%的(NH4)2SO4溶液作为沉淀剂,沉淀时间0.5 h,质量分数为0.02%的戊二醛作为交联剂,交联时间1 h。 通过比较酶学性质发现,ACE-CLEAs比游离ACE具有更好的温度稳定性及pH稳定性,且与游离ACE接近的Km值表明,ACE-CLEAs对底物的亲和力与游离酶几乎相当。     

胰蛋白酶交联聚体的制备及性质研究

交联酶聚体(CLEA)是一种新型的无载体固定化技术. 以胰蛋白酶为模型体系, 系统地研究了CLEA技术的制备工艺、应用条件、稳定性及结构形貌. (1)在制备工艺中考察了各步骤对CLEA酶活保留的影响, 重点分析了沉淀剂浓度、类型的影响, 结果表明100%乙醇是较为理想的沉淀剂; (2)在应用条件确定中测定CLEA的最适催化温度为70 ℃, 最适催化pH为9.0, 并解释了温度-活性、pH-活性曲线的漂移现象; (3)稳定性研究结果表明CLEA技术大幅提高了胰蛋白酶的热稳定性、溶剂稳定性, 且无酶泄漏; (4)在形貌表征中分别利用扫描电子显微镜、光学显微镜和激光粒度分析对CLEA的微观结构进行了多尺度研究, 着重讨论了其独特结构与优良特性间的关系. 本文所得结论为胰蛋白酶CLEA的应用提供基础数据, 并为CLEA技术应用于其它酶种提供参考.     

磁性壳聚糖微球的制备及其用作漆酶固定化载体

磁性壳聚糖微球的制备及其用作漆酶固定化载体;壳聚糖;固定化;漆酶;酶活力     

PVA固定化脲酶电极的制作及电极特性的研究

利用PVA低温下的物理交联固定酶,制成酶电极并在不同尿素浓度、pH值、温度等条件下进一步研究这种酶电极的响应特性.     

固定化脲酶的制备及应用

以壳聚糖作为载体,戊二醛作为交联剂对脲酶进行固定化。固定化的最适条件为:酶的偶联时间60min,戊二醛浓度0.5%,pH值7.0。对游离及固定化脲酶的酶学性质研究表明,酶促反应的最适pH均为7.0,最适温度分别为33℃和70℃。米氏常数分别为29.8mmol/L和13.9mmol/L。与游离酶相比,固定化酶的热稳定性和贮存稳定性更佳。应用固定化酶测定了试样中的微量组分。     

Preparation,Characterization and Optimization of Chitosan Nanoparticles as Carrier for Immobilization of Thermophilic Recombinant Esterase

Immobilization of biologically important molecules on myriad nano-sized materials has attracted great attention. Through this study, thermophilic esterase enzyme was obtained using recombinant DNA technology and purified applying one-step His-Select HF nickel affinity gel. The synthesis of chitosan was achieved from chitin by deacetylation process and degree of deacetylation was calculated as 89% by elemental analysis. Chitosan nanoparticles were prepared based on the ionic gelation of chitosan with tripolyphosphate anions. The physicochemical properties of the chitosan and chitosan nanoparticles were determined by several methods including SEM (Scanning Electron Microscopy), FT-IR (Fourier Transform Infrared Spectroscopy) and DLS (Dynamic Light Scattering). The morphology of chitosan nanoparticles was spherical and the nanospheres’ average diameter was 75.3 nm. The purified recombinant esterase was immobilized efficiently by physical adsorption onto chitosan nanoparticles and effects of various immobilization conditions were investigated in details to develope highly cost-effective esterase as a biocatalyst to be utilized in biotechnological purposes. The optimal conditions of immobilization were determined as follows; 1.0 mg/mL of recombinant esterase was immobilized on 1.5 mg chitosan nanoparticles for 30 min at 60°C, pH 7.0 under 100 rpm stirring speed. Under optimized conditions, immobilized recombinant esterase activity yield was 88.5%. The physicochemical characterization of enzyme immobilized chitosan nanoparticles was analyzed by SEM, FT-IR and AFM (Atomic Force Microscopy).     

Modified Polymer for Enzyme Immobilization and Characterization of Immobilized Urease

Abstract

Poly-γ-methyl-L-glutamate was modified by the introduction of -CC1₃ and /or -NH₂ groups. Two kinds of polymer were obtained, and applied to urease immobilization. The enzyme immobilized on polymer containing -CCl₃ (PCCl₃) pendant groups retained more activity than that containing -NH₂ (PNH₂). Stability of urease immobilized on PCCl₃ and PNH₂ was 17 days and 19 days respectively.     

Preparation and Characterization of Quaternized Chitosan Under Microwave Irradiation

For the first time, N-(2-hydroxyl) propyl-3-trimethyl ammonium chitosan chloride (HTCC) was prepared through a fast, easy and efficient method with the assistance of microwave irradiation, and the quaternized chitosan was also degraded via the microwave irradiation. A comparative study was performed by using the conventional heating method to prepare HTCC. The structure and property of the quaternized chitosan obtained by these two methods were characterized by GPC, XRD, FTIR, NMR, TG and elemental analysis. It was shown that quaternized chitosan was successfully prepared within 50 min via microwave irradiation method, while a much longer time of 6–7 h was needed with the conventional heating method. The substitutions both occurred on the C2 position of chitosan with the two different methods, and their HTCC products had weight average similar molecular weight (Mw), structure and thermal stability. The HTCC prepared by the microwave irradiation method had a little lower degree of substitution (DS) than those prepared via conventional heating with the same mole ratio (6:1) of the intermediate to chitosan. The degradation study showed that the Mw of HTCC decreased rapidly from 4.6 × 10⁵ to 1.1 × 10⁵ in 1 h under microwave irradiation, while it only decreased from 4.6 × 10⁵ to 2.1 × 10⁵in 1 h through conventional heating degradation. These results revealed that microwave irradiation is a more efficient and environment-friendly way to obtain the water-soluble chitosan derivatives and their degraded products.     

壳聚糖固定化乳酸脱氢酶的制备及酶学性质研究

以磁性壳聚糖作为载体,戊二醛作为交联剂,对乳酸脱氢酶(LDH)进行固定化.固定化的最适条件为:戊二醛浓度6%,pH值7.5,酶的偶联时间2 h.对游离及固定化LDH酶学性质的研究表明,酶促反应的最适pH值为9.2,最适温度分别为37℃和50℃,对乳酸的表观米氏常数分别为1.6 mmol/L和0.9 mmol/L.游离酶和固定化酶在40℃放置150 min后,其活力分别为最初的56.5%和76.1%.固定化酶在4℃贮存4周后,活力仍保留50%以上.固定化酶在室温下与底物重复反应6次后,活力仍保留60%以上,说明固定化酶具有较好的热稳定性、贮存稳定性和复用性.     

基于N-羧甲基壳聚糖钆基磁共振造影剂的制备及性能

文通过在DTPA配体上修饰N-羧甲基壳聚糖（NCMCS）增加配体的分子量,构建配合物Gd-（DTPA-NCMCS）.测定了Gd-（DTPA-NCMCS）及对照组Gd-DTPA与Gd-（DTPA-CS）不同浓度的弛豫时间T₁,拟合了配合物的纵向弛豫率r₁,结果表明Gd-（DTPA-NCMCS）纵向弛豫能力明显强于Gd-DTPA,也高于配合物Gd-（DTPA-CS）,浓度梯度的体外加权成像图清晰地观察到配合物Gd-（DTPA-NCMCS）比Gd-DTPA、Gd-（DTPA-CS）对应水溶液的信号更强。MTT法测定了Gd-（DTPA-NCMCS）的IC₅₀值为568 μmol·L^-1,表现出良好的生物相容性。Gd-（DTPA-NCMCS）可作为潜在的磁共振造影剂。     

交联壳聚糖膜的制备及其性能的研究

用环氧氯丙烷成功地制备出交联壳聚糖膜。用FTIR,XRD和SEM方法表征其结构,并测试了其力学性能。结果表明,壳聚糖在低温下只有氨基参与交联反应,反应温度高于40℃时,羟基才发生反应;环氧氯丙烷的交联作用显着提高了壳聚糖膜的抗张强度,并有效地降低了溶菌酶对其降解速率;该交联膜有望用作可控降解的生物医用材料。     

The Preparation,Characterization and Properties of Catalase Immobilized on Crosslinked Gellan

In the present paper, the reaction of chemical immobilization of catalase on a crosslinked macromolecular carrier of a polysaccharide structure (gellan) is studied. The influence of some reaction parameters (enzyme/carrier, activator/carrier ratios, duration) on the activity of enzymatic products is analyzed. The kinetics of the biocatalytic process, stability under different pH and temperature conditions, and the inhibitors effect were studied for the immobilized enzymes.     

微波辐射法制备N-亚水杨基壳聚糖Schiff碱及其吸附性能的研究

微波辐射法制备N-亚水杨基壳聚糖Schiff碱及其吸附性能的研究;壳聚糖;微波辐射;水杨醛;Schiff碱     

Preparation of Immobilized Metal Affinity Chromatographic Packings Based on Monodisperse Hydrophilic Non‐porous Beads and Their Application

Three hydrophilic immobilized metal affinity chromatographic packings for HPLC have been synthesized by chemical modification of 3.0 µm monodisperse non‐porous poly(glycidyl methacrylate‐co‐ethylenedimethacrylate) (P_GMA/EDMA) beads. The retention behavior of proteins on the metal ion chelated columns loaded with copper(II), nickel(II) and zin(II) ion was studied. The effect of pH on the protein retention was investigated on both the naked and metal ion chelated columns in the range from 4.0 to 9.0. Four proteins were quickly separated in 3.0 min with linear gradient elution at a flow rate of 3.0 mL/min by using the synthesized Ni²⁺‐IDA (iminodiacetic acid) packings. The separation time was shorter than other immobilized metal affinity chromatography reported in the literature. Purification of lysozyme from egg white and trypsin on the commercially available trypsin was performed on the naked‐IDA and Cu²⁺‐IDA columns, respectively. The purities of the purified trypsin and lysozyme were more than 92% and 95%, respectively.