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81.
82.
多孔硅球固定化木瓜蛋白酶的制备和性质 总被引:11,自引:0,他引:11
用载体交联法制备了多孔硅球固定化木瓜蛋白酶。考察了固定化时间、温度、pH值、给酶量和成二醛浓度对固定化木瓜蛋白酶活力的影响。研究了固定化木瓜蛋白酶的性质,并同溶液酶进行了比较。着重考察了固定化木瓜蛋白酶的热稳定性。所制得的固定化木瓜蛋白酶最适温育温度达到80℃,对底物酪蛋白的水解活力随温度的升高而增加,在90℃达到最高值;在70℃温育12小时后酶活力仍能保持高水平。 相似文献
83.
A sensitive and selective column adsorption method is proposed for the off-line preconcentration and determination of phthalic acid esters (PAEs), namely benzyl-butyl phthalate (BBP), di-n-butyl phthalate (DBP) and di-cyclohexyl phthalate (DCHP). The PAEs was preconcentrated on Saccharomyces cerevisiae immobilized on silica gel and then determined by high performance liquid chromatography (HPLC). Several parameters on the recovery of the analytes were investigated. Recoveries of BBP, DBP and DCHP were 100±2, 98±2 and 98±3%, respectively, at 95% confidence level under optimum conditions. The detection limits (3S/N) of BBP, DBP and DCHP were 3.2, 6.3 and 3.1 μg l−1, respectively. The capacity of the adsorbent was also examined and found to be 1.4 mg g−1 for BBP and DBP, and 3.6 mg g−1 for DCHP. S. cerevisiae immobilized on silica gel is suitable for repeated use without decreasing recovery up to about 25 adsorption-elution cycles. The proposed method was successfully applied to the determination of PAEs in river water with high precision and accuracy. 相似文献
84.
In this paper, the poly(acrylamide) hydrogel used to immobilize saccharomyces cerevisiae for asymmetric synthesis of R(-)-mandelic acid was prepared with free radical ploymerization in deionized water at room temperature under nitrogen atmosphere. The influence of the composition of hydrogel, loading amount of cells and culture conditions on the asymmetric synthesis was investigated. Results show that PAAm hydrogel is a feasible carrier for immobilization of cells which is a potential alternative method to prepare enantiomerically pure R(-)-mandelic acid. 相似文献
85.
Kiyoshi Shibata Hiroshi Kimura Akira Tsuchida Tsuneo Okubo Shohei Sato Kohji Yoshinaga 《Colloid and polymer science》2006,284(4):372-379
Electro-optic responses of colloidal crystals consist of poly(maleic anhydride-co-styrene)-modified silica spheres (P(MA-ST)/SiO2) in acetonitrile and the crystals immobilized with a gelator, N-benzyloxycarbonyl-L-isoleucilaminooctadecane (Z-L-Ile-C-18), are studied by reflected-light intensity measurements and time-resolved reflection spectroscopy. Application of
an alternating electric field deforms P(MA-ST)/SiO2 crystal lattices reversibly. The response waveforms from the crystals are dependent on the frequency and strength of the
applied electric field; similar dependencies have been qualitatively observed for the colloidal crystals consisting of polystyrene
or silica spheres in aqueous media in our previous studies. Both gelated and ungelated P(MA-ST)/SiO2 crystals change the reflection intensity, however, the amplitude is larger for the latter. The small response for the gelated
P(MA-ST)/SiO2 crystals is attributed to the higher elastic modulus (G). The G value of the gelated P(MA-ST)/SiO2 crystals in acetonitrile is estimated from the change in the inter-sphere distance to be 8.0 Pa, which is about 2.3 and 2.4
times larger than that for ungelated P(MA-ST)/SiO2 crystals in acetonitrile and colloidal silica crystals in aqueous media, respectively. 相似文献
86.
Immobilization of trypsin on porous glycidyl methacrylate beads: effects of polymer hydrophilization 总被引:2,自引:0,他引:2
The immobilization of trypsin at porous glycidyl methacrylate (GMA-GDMA) beads was investigated. In particular, the effects of surface modification of the beads through hydrophilic polymers on the amount protein immobilized and on the extent of retained activity after immobilization were adressed. Furthermore, immobilization at unmodified and hydrophilized beads from aqueous solution was compared to that from a water-in-oil microemulsion. It was found that the amount trypsin immobilized at the unmodified GMA-GDMA beads was significantly higher than that at hydrophilized GMA-GDMA beads. However, also the extent of specific activity loss after immobilization was larger for the unmodified than for the hydrophilized beads. Despite the latter, however, the total activity displayed by the hydrophilized beads was comparable to the unmodified beads at best. On the other hand, by peforming the immobilization from the microemulsion a high immobilization yield can be reached even for the hydrophilized beads, which also results in a higher degree of retained activity in the latter case than obtained for immobilization at the unmodified beads. Using this approach therefore resulted in the highest total activity of the trypsin-activated GMA-GDMA beads. 相似文献
87.
亲水性交联聚合物载体的合成及其固定化青霉素酰化酶 总被引:4,自引:0,他引:4
选用含环氧基团的甲基丙烯酸缩水甘油酯(GMA)和亲水性的N-乙烯吡咯烷酮(NVP)单体,以N,N′-亚甲基双丙烯酰胺(MBAA)为交联剂,甲酰胺作致孔剂,通过反相悬浮聚合技术成功合成了一系列大孔、珠状GMA-NVP-MBAA三元共聚物载体.N-乙烯吡咯烷酮介入共聚物体系,使共聚物载体具有较强的亲水性,有利于青霉素酰化酶的固定化.通过调节交联剂的用量和单体NVP与GMA的比例,可以调节共聚物载体的孔结构与表面性能.用合成的平均孔径为15.7nm、表面环氧基含量1.11mmol·g-1亲水性珠状载体固定青霉素酰化酶,固定化酶水解青霉素G钾盐的活性达491U·g-1;在4℃保存30d,活性保持不变.经4次使用后活性达到稳定(444U·g-1),再经14次使用后,活性没有明显变化. 相似文献
88.
《Electroanalysis》2004,16(22):1891-1895
An ISFET based enzymatic biosensor was developed for the determination of urea. Immobilization of urease was accomplished by the use of liquid mixture which contained vinylpyrrolidone, oligouretane metacrylate and oligocarbonate metacrylate and which can form a polymer under the influence of ultraviolet. The biosensor has the following characteristics: the linear field of responses is in the range of 0.05–20 mM, curve slope – 38 mV/pC, and response time 5–10 min. The increase of the temperature from 28 to 41 °C leads to 15% increase in the intensity of the response of the biosensor. The maximum response is observed at pH 6.0–6.5. At the increase of the NaCl concentration in solution up to 300 mM the biosensor response drops off and achieves half of its initial level. NH4Cl causes a stronger inhibition of enzyme activity comparing to NaCl. The results obtained with the developed biosensor correlate with the data of standard calorimetric methods. The intensity of the biosensor response decreases gradually during 40 days up to 80% of the initial level. The biosensors prepared with a fresh membrane or membrane preserved during 46 days at 2 °C gave similar responses in solution with an equal concentration of a substrate. It is concluded that the developed enzymatic biosensor is perspective for its clinical application for the determination of urea in blood and that the proposed method to prepare a selective biological membrane may be in a simple way included in integral technology of the semiconductor transducer manufacturing. 相似文献
89.
Bhavender P. Sharma Lorraine F. Bailey Ralph A. Messing 《Angewandte Chemie (International ed. in English)》1982,21(11):837-854
The immobilization of enzymes, cofactors and whole cells offers the potential of economical exploitation of biomaterial activity. Enzymes or cells can be immobilized by binding to carriers through adsorption or covalent linkages or by entrapment in a polymer matrix or microencapsulation. The development of immobilized biomaterial systems requires the involvement of biochemical, kinetics and reactor design principles. Examples of commercial scale applications of immobilized biomaterial systems are the production of a fructose-rich syrup from starch and the synthesis of L -amino acids by selective hydrolysis of N-acyl-D ,L -amino acids. 相似文献
90.
Kinetic studies and analytical application of cholesterol oxidase and peroxidase immobilized to synthetic polymer 总被引:2,自引:0,他引:2
A method for individual and simultaneous covalent immobilization of cholesterol oxidase and peroxidase to copolymer of acrylonitrile
with acrylamide is described. The effect of immobilization on the catalytic properties of the covalently bound enzymes was
studied. The immobilized enzymes showed no change in pH optima and an increase in temperature optima, activation energy, and
K
m
, compared to data received from experiments with soluble enzymes. A small glass column packed with immobilized multienzyme
complex was used to develop a method for manual determination of cholesterol in foodstuffs (e.g., in mayonnaise “Olinease”).
The method was characterized by high analytical precision (coefficient of variation = 2.67%). The results show high correlation
with those obtained by the Kageyama method (r=0.986). The method is economical (the enzyme-carrier conjugate may be used more than 300 times), precise, easy to perform,
and less time-consuming than the manual methods utilizing soluble enzymes. The established manual method can be proposed for
cholesterol determination in foodstuffs. 相似文献