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本文以大豆多糖为载体,分别用吸附法与戊二醛交联法固定化脲酶,测得了吸附法的最佳反应温度为65~75℃,pH为6.0~7.0。并对大豆多糖固定化脲酶柱式反应器的动态活性作了初步的测试。  相似文献   

3.
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。  相似文献   

4.
聚乙烯醇高含水胶固定化脲酶的研究   总被引:2,自引:0,他引:2  
冷冻-部分脱水法制成的聚乙烯醇高含水胶固定化脲酶活力收率及对脲素的分解能力明显高于聚丙烯酰胺囱定化脲酶,稳定性相近;用低浓度的戊二醛后处理,提高了固定化酶的稳定性。  相似文献   

5.
醋酸纤维素膜固定化脲酶的研究   总被引:9,自引:0,他引:9  
酶固定膜反应器兼具有反应和分离两种功能,是酶工程领域中较活跃的研究课题.随着酶固定化技术和水平的提高,各种固定化酶生物反应器不断涌现,其中以采用固定化脲酶技术制作的人工肾最为成功.  相似文献   

6.
固定化酶是束缚在一定空间内仍能进行其特有反应的生物催化剂,近年来对于固定化酶的研究非常活跃[1-2].目前酶的固定化方法主要有吸附法,交联法,包埋法以及化学共价法,这些方法各有特点[3-5],应用领域多为食品工业、制药、医学治疗等方面.  相似文献   

7.
陈佩琴  何斌 《分析化学》1993,21(10):1135-1138
报道了一种新的L-氨基酸氧化酶电极,这种酶电极系由氨气敏电极和以氨基化玻璃布为载体的酶膜所组成;研究了固定化条件对酶膜活性的影响以及底物浓度、温度和pH对电极响应特性的影响。该电极在6.0×10^-5~4.0×10^-3mol/L的底物浓度范围内呈良好的线性关系,检测下限为5.0×10^-5mol/L。在最宜条件下,酶电极具有良好的稳定性。  相似文献   

8.
流动注射;尿素测定;壳聚糖固定化脲酶的制备和废水中的尿素分析  相似文献   

9.
以环氧氯丙烷活化的磁性壳聚糖微球作为载体,对脲酶进行固定化研究。结果表明, 在25℃时, 活化磁性壳聚糖微球对脲酶的固定化在2h时就达到了最大值,固定化酶和自由酶的最适温度都在65℃左右,自由酶和固定化酶的米氏常数Km值分别为0.042mol/L和0.008mol/L,固定化酶的Km降低了5倍。  相似文献   

10.
提出了一种新的酶固定化方法, 即通过甲醇处理, 使蚕丝素蛋白膜的构象由random coil向β-sheet发生根本性的变化, 从而将酶固定在β-sheet所特有的分子间氢键中。利用此方法所制成的脲酶电极, 在合适的操作条件下, 各项响应指标均令人满意, 并且脲酶的耐温性能被大大提高, 电极的有效使用寿命长达三个月以上。此种酶固定化方法原则上能够应用于其他不破坏蚕丝素蛋白分子结构的可溶性酶。  相似文献   

11.
《Electroanalysis》2002,14(23):1644-1647
The activity of urease varies by its redox reaction. Active urease has an SH group that is essential to exhibit its activity, however, oxidation agents such as quinone compounds can oxidize the SH group in urease and a S–S bond is produced, resulting in the loss of enzyme activity. The reduction potential of cystine was almost the same as that of the recovery of urease activity. In this work, it has been found that the SH group of urease can be oxidized by not only chemical reaction but also by the direct electrode oxidation of urease and the produced S–S bond can be reduced to SH group by chemical and electrode reactions, and the original enzyme activity is recovered. This research shows that the regulation of urease activity is easily possible by changing the electrode potential of the porous carbon felt immobilized urease. The variation of urease activity was monitored by ammonia or carbon dioxide electrode equipped with the urease immobilized carbon felt, and the ammonia or carbon oxide generated from urea can transfer through the carbon felt to reach the each gas permeable membrane. The combination of gas electrode with porous conducting material such as carbon can supply the novel device for the electrochemical investigation of enzyme activity.  相似文献   

12.
A large oxidation current can be observed when ammonium carbamate aqueous solution is electrolyzed using a glassy carbon electrode (GCE) at a potential exceeding 1.0 V vs. Ag/AgCl and amino groups are introduced at the surface of the GCE. Aminated GCE exhibits the electrocatalytic activity of the oxidation of ammonium carbamate that is produced from urea as an intermediate product of urease reaction, and a distinct oxidation current is observed when the aminated GCE is used to oxidize the urea in the urease solution. A novel amperometric determination method to detect urea has been developed. This method is based on the electrooxidation of carbamic acid produced during urease reactions. Urease is immobilized to polymaleimidostyrene (PMS) coated on the insulated amorphous carbon sheet set on the aminated GCE surface. A good linear relationship is observed between urea concentration and the electrolytic current of the urease‐immobilized electrode in the concentration range from 0.5 mM to 21.0 mM. The proposed urea biosensor has an effective merit in that the interference resulting from ammonia and pH change caused by the urease reaction can be eliminated, differing from conventional urea biosensors.  相似文献   

13.
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。  相似文献   

14.
《Analytical letters》2012,45(11):987-1001
Abstract

The application of an enzyme thermistor device in a simple and accurate procedure for the determination of serum urea is described. The enzyme thermistor measures the heat produced when urea is passed through a small column containing immobilized urease. The stability and sensitivity as well as the performance with clinical serum samples of the system is evaluated. Advantages are the simplicity, the low enzyme cost and the insensitivity to the optical properties of the sample and interfering substances, which may affect the commonly used assay procedures.  相似文献   

15.
Nickel oxide nanoparticle (NiO?NP) and polypyrrole (PPy) composite were deposited on a Pt electrode for fabrication of a urea biosensor. To develop the sensor, a thin film of PPy?NiO composite was deposited on a Pt substrate that serves as a matrix for the immobilization of enzyme. Urease was immobilized on the surface of Pt/PPy?NiO by a physical adsorption. The response of the fabricated electrode (Pt/PPy?NiO/Urs) towards urea was analyzed by chronoamperometry and cyclic voltammetry (CV) techniques. Electrochemical response of the bio‐electrode was significantly enhanced. This is due to electron transfer between Ni2+ and Ni3+ as the electro‐catalytic group and the reaction between polypyrrole and the urease‐liberated ammonium. The fabricated electrode showed reliable and demonstrated perfectly linear response (0.7–26.7 mM of urea concentration, R2= 0.993), with high sensitivity (0.153 mA mM?1 cm?2), low detection of limit (1.6 μM), long stability (10 weeks), and low response time (~5 s). The developed biosensor was highly selective and obtained data were repeatable and reproduced using PPy‐NiO composite loaded with immobilized urease as urea biosensors.  相似文献   

16.
《Analytical letters》2012,45(7):525-540
Abstract

A sensitive method for the rapid determination of activities of soluble or immobilized enzymes, based on the electrochemical detection of hydrogen peroxide is described. Kinetic studies (Vmax and KM determinations) can be performed for all H2O2 generating enzymes (i.e. most of the oxidases) using an amperometric probe with a platinum anode at a fixed potential.

When associated with an immobilized glucose oxidase membrane, this sensor constitutes a glucose electrode and the activity of any hydrolase which releases glucose can be measured. There is no need for other auxiliary enzymes and no preincubation step is required. The possibility to carry out continuous analysis constitutes the main advantage of the described method.  相似文献   

17.
在电位型尿素酶电极的组装过程中,要求尽量不改变尿素酶的构型和构象,使其在酶膜中保持其自然状态,从而可获得较高的酶活力.用pH玻璃电极作原电极,将尿素酶固定在其表面,常用的有戊二醛交联法[1]和各种聚合物膜法[2~4].本文利用60%季铵化的聚(4-乙...  相似文献   

18.
聚苯胺葡萄糖氧化酶电极的催化过程   总被引:1,自引:0,他引:1  
陆寿蕴  李诚芳 《分析化学》1993,21(8):946-949
用电化学方法固定在直径为0.5mm铂丝上的聚苯胺(PANI)葡萄糖(GOD)电极对葡萄糖有催化氧化作用.在0~-0.6V(vs.SCE)的电极范围内,在电极的循环伏安曲线上观察到与葡萄糖浓度有关的氧的还原峰和GOD还原态的氧化峰,用此GOD还原态的氧化峰电流可定量检测葡萄糖的浓度。本文提出在PANI电极上存在着酶反应氧化还原电荷直接传递的可能性。  相似文献   

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