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51.
In this study we describe the activation with chloroformates of Trisacryl-GF-2000, a new synthetic gel support that is stable,
hydrophilic, and contains large amounts of hydroxyl groups available for activation.
Of all the reagents tested, the activation withN-hydroxysuccinimide-chloroformate andp-nitrophenylchloroformate in organic solvents provides the best activation yield and subsequent coupling. When Trisacryl was
activated in acetone with the chloroformates in the presence of 4-dimethylaminopyridine as base and catalyst, up to 30% of
the hydroxyl groups, (i.e., 1/repeating unit) could be activated. Amino-containing ligands and proteins could be coupled to
these carriers at pH 8 or higher. For better results in affinitychromatographic applications, spacers of ε-amino caproic acid
or diaminohexane were introduced. The efficacy of these columns was demonstrated by purification of enzymes, antibodies, and
antigens. The performance of these new columns were compared with that of Sepharose columns activated in various ways. In
every case, the properties of the Trisacryl support proved superior with particular reference to the purity of the product
obtained. 相似文献
52.
The Morita-Baylis-Hillman reaction of cyclohexenone and p-nitro benzaldehyde is catalyzed by carrier proteins such as serum albumins or enzymes such as certain lipases, conversion of up to 35% and enantioselectivities of up to 19% being observed. 相似文献
53.
葡萄糖氧化酶电极的制备 总被引:5,自引:0,他引:5
葡萄糖氧化酶电极的制备于秀娟,周定,郭京华(哈尔滨工业大学应用化学系哈尔滨150006)关键词葡萄糖氢化酶,酶电极,制备酶与电极的连接即酶膜的形成是制备酶电极的关键步骤之一,有许多种方式[1~9].葡萄糖氧化酶(GOD)电极是研究较早和报道较多的一类... 相似文献
54.
Vlatakis G. Skarpelis G. Stratidaki I. Bouriotis V. Clonis Y. D. 《Applied biochemistry and biotechnology》1987,15(3):201-212
The resolution of restriction endonucleases from the same microorganism is conventionally achieved by lengthy fractionation
protocols. We now report effective single-step procedures that exploit dye-ligand chromatography for the resolution and purification
of restriction enzymes. After suitable initial screening, we demonstrated that resolution of two restriction activites can
be achieved in one chromatographic step, and further purification can subsequently be effected using selected dye-adsorbents.
Accordingly, we resolved in one step, Hpa I from Hpa II, Hind II from Hind III, and Sac I from Sac II. Furthermore, a three-step
Chromatographic procedure has been developed to purify EcoRV suitable for commercial exploitation, as judged by the “overdigestion”
and “cut-ligate-recut” quality control tests. 相似文献
55.
微胶囊固定化过氧化氢酶 总被引:5,自引:1,他引:5
以乙基纤维素为膜材,用液中干燥法将过氧化氢酶微胶囊化,研究了温度和pH值对酶活性的影响及过氧化氢和尿素对固定化酶的抑制作用,由于乙基纤维素膜的保护,明胶对pH值的缓冲作用及明胶与尿素的反应,明显地扩大了酶对温度和pH值的适应性,降低了酶活性抑制剂的影响。 相似文献
56.
本文报道了金属置换碳酸酐酶的活性研究。指出了离子半径的大小,配位构型的要求,热力学稳定性和配位体交换动力学是决定各种金属置换碳酸酐酶活性的重要因素。并用金属置换法测定了Ni(Ⅱ)与apo-CA形成金属酶的反应动力学速率常数。在25℃、pH7.47时,K_(fNi(Ⅱ)-CA)为14.01·mol~(-1)sec~(-1),较之于Zn(Ⅱ)-CA的速率常数小10~3倍。Cd(Ⅱ)-CA的生成动力学为一快反应,采用配位竞争法,在邻菲罗啉配位剂存在下,测得了Cd(Ⅱ)-CA生成动力学的条件速率常数在o-phen浓度为1.96和2.92×10~(-4)mol·1~(-1)时分别为137.1和114.61·mol~(-1)sec~(-1)。这一研究提示了镉对生物体的毒性作用的一个重要方面是使锌酶失活。而配位剂的存在大大降低了镉与脱辅基锌酶的反应动力学,这也是它们显示解毒功能所在。 相似文献
57.
Zanin Gisella M. Neitzel Ivo De Moraes Flavio F. 《Applied biochemistry and biotechnology》1993,(1):477-489
The axial dispersion of a liquid fluidized bed of controlled pore silica (CPS) particles has been determined by the pulse
tracer method. The CPS used was the same as for enzyme immobilization, having an average diameter of 0.436 mm and mean pore
size of 37.5 nm. The fluidization liquid is α-amylase liquefied manioc starch, 30% w/v, 45°C pH=4.5. Nominal bed porosities
tested were 0.7 and 0.8. The results show that the axial dispersion coefficient increases with greater superficial liquid
velocities. Various available correlations tested disagree with each other to a large extent and are unable to represent collected
experimental data. 相似文献
58.
59.
Lecithin/propanol-based microemulsions used as media for a cholesterol oxidase-catalyzed reaction 总被引:1,自引:0,他引:1
Reverse micelles, Winsor III and IV systems were examined as reaction media for the enzymatic conversion of cholesterol to cholestenone by cholesterol oxidase at 298.2 K. The micelles and the microemulsions, stabilized by soybean lecithin and ethanol or 1-propanol as cosolvent, were characterized with respect to phase behavior and distribution of 1-propanol between the phases of the Winsor III systems. The used oils were dodecane, tetradecane, and hexadecane. The Winsor IV systems and the surfactant-rich phase in the Winsor III systems exhibit bicontinuous structures. The reaction yield for the enzymatic conversion performed in a Winsor IV system was much higher than in a Winsor III system or in reverse micelles. 相似文献
60.