共查询到17条相似文献,搜索用时 156 毫秒
1.
固定化木瓜蛋白酶活性的X-射线微区分析 总被引:1,自引:0,他引:1
利用X-射线微区分析,对共价法得到的固定化木瓜蛋白酶的活性进行了分析;Na-苯甲酰-L-精氨酰胺盐酸盐作为底物,FeCl_3作为捕捉剂,底物经木瓜蛋白酶催化分解产生L-精氨酸及氨,后者和捕捉剂反应产生沉淀,可以确定固定化木瓜蛋白酶的催化活性部位。结果表明:以大孔结构吸附树脂为固定化酶载体,酶活较高,木瓜蛋白酶分布较均匀。并得到了固定化木瓜蛋白酶的活性定位的最佳条件。并对不同结构载体固定化木瓜蛋白酶的活性进行了研究。 相似文献
2.
壳聚糖固定化脲酶活性的X射线微区分析 总被引:2,自引:0,他引:2
利用X射线微区分析方法,对壳聚糖固定化脲酶活性进行了定位分析。筛选出了能捕捉固定化脲酶活性部位的捕捉剂BaCl2,以尿素作为底物,在Tris-HCl缓冲溶液中,底物经固定化脲酶催化产生NH3和CO2,后者和捕捉剂反应生成BaCO3沉淀, 沉积在固定化脲酶的催化活性部位,借此确定其催化活性部位。结果表明BaCl2可用做固定化脲酶活性定位的捕捉剂。同时得到了壳聚糖固定化脲酶活性定位的最佳实验条件。 相似文献
3.
利用X射线微区分析, 对二氧化硅溶胶-凝胶包埋于普鲁士蓝修饰玻碳电极上的葡萄糖氧化酶的活性进行了分析; 以Ce(NO3)3为捕捉剂, 底物葡萄糖经葡萄糖氧化酶作用产生过氧化氢,后者与捕捉剂反应生成沉淀于酶的活性部位。从X射线微区分析结果表明: 酶电极表面固定化酶的分布均匀,且保存较高的酶活,从微观的角度说明了酶电极的性能与酶电极表面酶活分布的关系。此法制备的葡萄糖氧化酶电极具有较高的灵敏度,稳定性,这与电化学测试结果是一致的。 相似文献
4.
5.
6.
7.
8.
研究3种不同类型的表面活性剂,对聚乙烯醇固定化乙酰胆碱酯酶(PVA-AChE)活性的影响.选用十六烷基三甲基溴化铵(CTAB)作酶促反应体系的增敏和终止剂,0.6g·L-1 CTAB使酶促反应增敏27%,5g·L-1 CTAB对酶促反应终止作用良好,优化了实验条件,初探了作用机理.用建立的方法测定马拉硫磷的检出限为1.8×10-9g·L-1,线性范围为8-160ng·mL-1,11次测定马拉硫磷(50ng· mL-1)的相对标准偏差为2.8%;用于测定小白菜样品中马拉硫磷的回收率为88.8%-95.6%. 相似文献
9.
一种可用于酚类化合物检测的酶传感器 总被引:1,自引:0,他引:1
利用溶胶-凝胶法将辣根过氧化物酶(HRP)固定化于二氧化硅凝胶网络中构建了可用于酚类化合物检测的酶传感器。对二氧化硅载体材料进行了结构表怔。二氧化硅多孔材料的平均孔径为2.95 nm。孔径小于5 nm占总数的84.068%。由于辣根过氧化物酶的分子尺寸远远大于二氧化硅的凝胶网络的平均孔径,因此不会泄露到溶液中去,而尺寸较小的底物可以发生反应。包埋的辣根过氧化物酶在H2O2的存在下,能够催化氧化苯酚与4-氨基安替比林反应生成醌亚胺有色化合物。通过紫外-可见光谱测定醌亚胺有色化合物的吸光度,即可以确立苯酚的含量。对于象含氯酚类的重要污染物, 如邻氯酚、间氯酚、2,4-二氯酚,这种方法也同样适用。此外,对多次测定以后的酶的活性下降的问题进行了讨论,结果表明酶传感器可以重复使用10次以上。但响应时间会变长。 相似文献
10.
一种用于化学分析的新方法——荧光毛细分析法 总被引:7,自引:2,他引:5
提出了一种新方法——荧光毛细分析法(fluorescence capillary analysis,FCA)。FCA法用毛细管做为荧光反应和测定的载体,实现了微量样品的微量分析。本方法特点是操作简便,试样、试剂用量少(<10 μL);毛细管及毛细生物反应器可循环使用,测定成本极低,能大大减少环境污染。该研究基于FCA法测定了核黄素、还原型辅酶Ⅰ,基于固定化酶FCA法测定了酒中乙醇,基于固定化核苷酸探针基因FCA法测定了靶核苷酸,都初步得到了满意的结果。FCA法将可以应用到临床医学、药学、生化、染料工业、发酵食品、环境检测等各个领域中。 相似文献
11.
In this work, Au–Ag nanoparticles (Au–Ag-bi-MNPs) have been prepared on amine functionalized Si-MCM-41 (NH2–Si-MCM-41) particles through a reduction of AgNO3 and HAuCl4 by NaBH4 at ambient conditions. Au–Ag-bi-MNPs loaded on the NH2–Si-MCM-41, provide a good biocompatible surface for immobilization of the enzyme alkaline protease. This immobilization, presumably due to bonding between core shell nanoparticles and OH in serine 183 in alkaline protease seems to be of an ionic exchange nature. We found that the alkaline protease immobilized on the Au–Ag-bi-MNPs/Si-MCM-41 is an active biocatalyst, stable at different pH and temperature. The bio catalytic activity of free alkaline protease in solution was 64 U/mg (Units per milligram), whereas that of the alkaline protease immobilized on Au–Ag-bi-MNPs/Si-MCM-41 was 75 U/mg. This improvement of the biocatalytic activity may be due to a really increased activity per molecule of immobilized enzyme or to a purification of the enzyme. The alkaline protease molecules immobilized on the (Au–Ag)/ NH2-MCM-41 surface retained as much as 80% of the catalytic activity recorded at pH=8, and showed significant catalytic activity of alkaline protease in the bioconjugate material. The biocatalytic materials were easily separated from the reaction medium by mild centrifugation and exhibits excellent reuse and stability characteristics over four successive cycles. The optimum temperature ranged from 35 °C–55 °C and pH=8 for bioactivity of the alkaline protease in the assembly system was observed to be higher than that of the free enzyme in solution. The enzyme biocatalytic activity was monitored by UV-visible spectroscopy. Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM) and dispersive analysis of X-RAY (EDAX) were used to characterize the size and morphology of the prepared materials. 相似文献
12.
Gui-yin Li Zhi-de Zhou Ke-long Huang 《Journal of magnetism and magnetic materials》2010,322(24):3862-588
A novel and efficient immobilization of yeast alcohol dehydrogenase (YADH, EC1.1.1.1) from Saccharomyces cerevisiae has been developed by using the surface functionalization of chitosan-coated magnetic nanoparticles (Fe3O4/KCTS) as support. The magnetic Fe3O4/KCTS nanoparticles were prepared by binding chitosan alpha-ketoglutaric acid (KCTS) onto the surface of magnetic Fe3O4 nanoparticles. Later, covalent immobilization of YADH was attempted onto the Fe3O4/KCTS nanoparticles. The effect of various preparation conditions on the immobilized YADH process such as immobilization time, enzyme concentration and pH was investigated. The influence of pH and temperature on the activity of the free and immobilized YADH using phenylglyoxylic acid as substrate has also been studied. The optimum reaction temperature and pH value for the enzymatic conversion catalyzed by the immobilized YADH were 30 °C and 7.4, respectively. Compared to the free enzyme, the immobilized YADH retained 65% of its original activity and exhibited significant thermal stability and good durability. 相似文献
13.
Hai Wang Jun HuangChao Wang Dapeng LiLiyun Ding Yun Han 《Applied Surface Science》2011,257(13):5739-5745
Aminated-CoFe2O4/SiO2 magnetic nanoparticles (NPs) were prepared from primary silica particles using modified StÖber method. Glucose oxidase (GOD) was immobilized on CoFe2O4/SiO2 NPs via cross-linking with glutaraldehyde (GA). The optimal immobilization condition was achieved with 1% (v/v) GA, cross-linking time of 3 h, solution pH of 7.0 and 0.4 mg GOD (in 3.0 mg carrier). The immobilized GOD showed maximal catalytic activity at pH 6.5 and 40 °C. After immobilization, the GOD exhibited improved thermal, storage and operation stability. The immobilized GOD still maintained 80% of its initial activity after the incubation at 50 °C for 25 min, whereas free enzyme had only 20% of initial activity after the same incubation. After kept at 4 °C for 28 days, the immobilized and free enzyme retained 87% and 40% of initial activity, respectively. The immobilized GOD maintained approximately 57% of initial activity after reused 7 times. The KM (Michaelis-Menten constant) values for immobilized GOD and free GOD were 14.6 mM and 27.1 mM, respectively. 相似文献
14.
建立利用大孔吸附树脂吸附法结合氯仿溶解法分离混合二元酸中丁二酸、戊二酸和己二酸的方法并使其分别达到一定的纯度和收率。首先利用LC-ESI-MS方法测定了混合二元酸中丁二酸、戊二酸和己二酸含量,然后通过对3种大孔吸附树脂AB-8、NKA-Ⅱ和D-380吸附性能的综合比较,筛选出AB-8大孔吸附树脂作为吸附剂,以水、10%、30%和95%乙醇水溶液作为洗脱剂对混合二元酸进行初步的分离,得到A、B、C和D4个分离部位,再利用氯仿对二元酸溶解度的不同,将该4个分离部位在55℃水浴恒温分别用氯仿进行溶解、过滤,通过LC-ESI-MS方法测定确定相应部位的组成情况后进行适当合并,最终达到将混合二元酸分离的目的。丁二酸纯度:70.3%,收率:92.7%;戊二酸纯度:94.1%,收率89.2%;己二酸纯度:90.7%,收率:88.7%。 相似文献
15.
The ultrasound assisted purification of asparagus polyphenols by adsorption and desorption on the macroporous resins was investigated. The ultrasound within the selected intensities (12–120 W) and temperatures (25–35 °C) increased the adsorption and desorption capacities of asparagus polyphenols on D101 resins. Higher ultrasound intensity (120 W) and lower temperature (25 °C) benefited the adsorption process and the adsorption capacity of total polyphenols after ultrasound was 3.95 mg/g, which was 2 times than that obtained after shaking at 120 rpm. Meanwhile, ultrasound can significantly shorten the equilibrium time and the adsorption process of asparagus polyphenols could be well described by Pseudo-second order model and Freundlich model. Stereoscopic microscope was first used to investigate the microstructure characterization of resins, indicating that ultrasound mainly enhanced the surface roughness of resins. Interestingly, rutin possessed the highest adsorption capacities and ferulic acid had the highest the desorption capacities among the studied individual polyphenols. The obtained results evidenced on a progressive insight of application of ultrasound assisted resins for purification of asparagus polyphenols. 相似文献
16.
在评价硝化抑制剂的作用效果时, 需要确认此硝化抑制剂对反硝化过程有无影响。而对此过程的确认是通过反硝化酶活性(denitrifying enzyme activity, 简写为DEA)的测定来实现的。采用加入同位素标记底物结合同位素质谱仪来测定新型硝化抑制剂3,4-二甲基吡唑磷酸盐作用下的反硝化酶活性。结果表明, 此新型方法能够较准确的测定培养体系中的N2O的浓度,与气相色谱法的测定结果具有良好的相关性(MSN2O=-0.45+1.03GCN2O,R2=0.995)。通过测定15N2O和15N2的丰度能够较好的区分2种反硝化酶活性(硝酸还原酶和N2O还原酶), 且克服了传统测定中需要加入乙炔作为酶抑制剂的弊端。对DMPP作用下土壤反硝化酶的测定表明,DMPP对反硝化酶无显著影响, 说明DMPP在使用中不会影响土壤中的反硝化过程。 相似文献
17.
Weiwei Zhu Yimei Zhang Chen Hou Duo Pan Jianjun He Hao Zhu 《Journal of nanoparticle research》2016,18(2):32
This paper reported an immobilization of Candida rugosa lipase (CRL) onto PAMAM-dendrimer-grafted magnetic nanoparticles synthesized by a modified solvothermal reduction method. The dendritic magnetic nanoparticles were amply characterized by several instrumental measurements, and the CRL was covalently anchored on the three generation supports with glutaraldehyde as coupling reagent. The amount of immobilized enzyme was up to 150 mg/g support and the factors related with the enzyme activity were investigated. The immobilization of lipase improved their performance in wider ranges of pH and temperature. The immobilized lipase exhibited excellent thermal stability and reusability in comparison with free enzyme and can be reused 10 cycles with the enzymatic activity remained above 90 %. The properties of lipase improved obviously after being immobilized on the dendritic supports. The inactive immobilized lipase could be regenerated with glutaraldehyde and Cu2+, respectively. This synthetic strategy was facile and eco-friendly for applications in lipase immobilization. 相似文献