共查询到20条相似文献,搜索用时 6 毫秒
1.
Rapid and high-capacity immobilization of enzymes based on mesoporous silicas with controlled morphologies 总被引:1,自引:0,他引:1
Fan J Lei J Wang L Yu C Tu B Zhao D 《Chemical communications (Cambridge, England)》2003,(17):2140-2141
Very rapid (< 10 min to reach equilibrium) and high-capacity (up to 533 mg g(-1)) immobilization of enzymes within mesoporous silica has been achieved by finely tuning their morphologies. 相似文献
2.
Immobilizing biocomponents on solid surfaces is a critical step in the development of new devices for future biological, medical, and electronic applications. Therefore, numerous integrated films were recently developed by immobilizing different proteins or enzymes on electrode surfaces. In this work, hemeproteins were safely immobilized onto macroporous nickel-based electrodes while maintaining their functionality.Such modified electrodes showed interesting pseudo-capacitive behavior. Among hemeproteins, hemoglobin(Hb) film has a higher electrochemical performance and greater charge/discharge cycling stability than myoglobin(Mb) and cytochrome C(Cyt C). The heme group in an alkaline medium could induce the formation of superoxides on the electrode surface. These capacitive features of hemeprotein-Ni electrode were related to strong binding sites between hemeproteins and porous Ni electrode, the accumulation of superoxide or radicals on the Ni surface, and facile electron transfer and electrolyte diffusion through the three-dimensional macroporous network. Thus, these new protein-based supercapacitors have potential use in free-standing platform technology for the development of implantable energy-storage devices. 相似文献
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介孔分子筛在生物酶固定化中的应用 总被引:7,自引:0,他引:7
综述了MCM-48和SBA-15等新型介孔分子筛用生物酶固定化载体研究的新进展。介孔分子筛由于拥有巨大的比表面积(~1000m2/g)、纳米尺寸孔道(2~50m)和较大的孔容(~1.0 cm3/g),因此以分子筛为载体利用物理吸附制备的固定化酶呈现出高的催化活性,但固定化酶操作稳定性较低,在使用过程中部分酶分子发生了脱落,其原因是分子筛表面自由的硅羟基通过物理吸附或氢键作用固定酶分子。借助介孔分子筛自身的自由硅羟基在表面嫁接-COOH、-NH2、-CH=CH2等有机官能团来构筑酶固定化的微环境,改善酶分子和载体的亲和作用,提高固定化酶的活性。目前,利用有机官能团功能化介孔分子筛固定化酶是研究发展的趋势。 相似文献
5.
Calmodulin-mediated reversible immobilization of enzymes 总被引:1,自引:0,他引:1
Daunert S Bachas LG Schauer-Vukasinovic V Gregory KJ Schrift G Deo S 《Colloids and surfaces. B, Biointerfaces》2007,58(1):20-27
This work demonstrates the use of the protein calmodulin, CaM, as an affinity tag for the reversible immobilization of enzymes on surfaces. Our strategy takes advantage of the of the reversible, calcium-mediated binding of CaM to its ligand phenothiazine and of the ability to produce fusion proteins between CaM and a variety of enzymes to reversibly immobilize enzymes in an oriented fashion to different surfaces. Specifically, we employed two different enzymes, organophosphorus hydrolase (OPH) and beta-lactamase and two different solid supports, a silica surface and cellulose membrane modified by covalently attaching a phenothiazine ligand, to demonstrate the versatility of our immobilization method. Fusion proteins between CaM-OPH and CaM-beta-lactamase were prepared by using genetic engineering strategies to introduce the calmodulin tail at the N-terminus of each of the two enzymes. In the presence of Ca(2+), CaM adopts a conformation that favors interaction between hydrophobic pockets in CaM and phenothiazine, while in the presence of a Ca(2+)-chelating agent such as EGTA, the interaction between CaM and phenothiazine is disrupted, thus allowing for removal of the CaM-fusion protein from the surface under mild conditions. CaM also acts as a spacer molecule, orienting the enzyme away from the surface and toward the solution, which minimizes enzyme interactions with the immobilization surface. Since the method is based on the highly selective binding of CaM to its phenothiazine ligand, and this is covalently immobilized on the surface, the method does not suffer from ligand leaching nor from interference from other proteins present in the cell extract. An additional advantage lies in that the support can be regenerated by passing through EGTA, and then reused for the immobilization of the same or, if desired, a different enzyme. Using a fusion protein approach for immobilization purposes avoids the use of harsh conditions in the immobilization and/or regeneration steps, which could cause inactivation of the immobilized enzyme. Moreover, we have demonstrated that the CaM affinity tag allows immobilization of enzymes on a variety of surfaces without compromising their enzymatic activity substantially; for example, the immobilized OPH retained more than 80% of the activity of the free enzyme. Our results with beta-lactamase showed the feasibility of using a phenothiazine surface in several consecutive loading and regeneration cycles. This can be advantageous when expensive and/or difficult to obtain immobilization surfaces have to be employed; the immobilization surface could be reused to immobilize the same or a different enzyme using the CaM affinity tail. We also determined that the phenothiazine-modified silica particles are stable for long periods of time, i.e., up to 2 years when stored at 4 degrees C. It is envisioned that this type of reversible immobilization may find applications in the development of reversible, reusable biosensors and bioreactors endowed with the additional advantage that the biological element at the surface of the sensor or bioreactor could be replaced under mild conditions when needed to sense or process a different target molecule. 相似文献
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V. V. Yanishpol'skii G. V. Lyubinskii V. A. Tertykh 《Theoretical and Experimental Chemistry》1989,25(6):690-693
We have studied the process of preparing a silica support with anhydride groups. The process includes introducing vinyl groups onto the surface with subsequent radiative copolymerization of maleic anhydride with them. The maximum degree of grafting of the functional groups of the matrix occurs for -irradiation doses of 20–30 Mrad. The carboxyl groups grafted to the aerosil surface, formed upon hydrolysis of the support, can be converted to anhydride by calcination at 403 K for 3 h.Translated from Teoreticheskaya i Éksperimental'naya Khimiya, Vol. 25, No. 6, pp. 750–753, November–December, 1989. 相似文献
8.
Luan Qian Zhang Hao Lei Yujie Cai Yixin Bao Yuping Li Yan Tang Hu Li Xiuting 《Cellulose (London, England)》2021,28(9):5735-5744
Cellulose - In this study, regenerated cellulose-based macrogels with abundant carboxyl groups and interconnected microporous structures were synthesized from cellulose fibers (CFs)/cellulose... 相似文献
9.
Yamamoto T Endo A Inagi Y Ohmori T Nakaiwa M 《Journal of colloid and interface science》2005,284(2):614-620
The accuracy of thermoporometry (TPM) in terms of the characterization of SBA-15 is examined based on a model that classifies the water in the mesopores into two different types: freezable pore water, which can form cylindrical ice crystals, and nonfreezable pore water, which cannot undergo a phase transition during a differential scanning calorimetry (DSC) measurement. Applying the empirical relationship between the sizes of the ice crystals formed in the mesopores and the solidification temperature of the freezable pore water to a thermogram (a recording of the heat flux during the solidification of the freezable pore water) yielded a size distribution of the ice crystals. The size of the ice crystals increased slightly with repetitive freezing, indicating that the mesopores were enlarged by formation of the ice crystals. Adding the thickness, t(nf), of the nonfreezable pore water layer to the ice crystal-size distribution calculated from the thermogram allowed for the determination of the porous properties of SBA-15. The porous properties attained from TPM experiments were compared with the results attained through the combination of Ar gas adsorption experiments and nonlocal density functional theory (NLDFT) analysis. The porous properties determined by TPM were confirmed to be quite sensitive to the t(nf) value. 相似文献
10.
Versatile mesoporous carbonaceous materials for acid catalysis 总被引:1,自引:0,他引:1
Budarin VL Clark JH Luque R Macquarrie DJ 《Chemical communications (Cambridge, England)》2007,(6):634-636
Starbon mesoporous materials were synthesized after pyrolysis of expanded starch and subsequently functionalised with sulfonated groups, providing highly active and reusable materials in various acid catalysed reactions. 相似文献
11.
Ordered mesoporous inorganic non-oxide materials attract increasing interest due to their plenty of unique properties and functionalities and potential applications. Lots of achievements have been made on their synthesis and structural characterization, especially in the last five years. In this critical review, the ordered mesoporous non-oxide materials are categorized by compositions, including non-oxide ceramics, metal chalcogenides, metal nitrides, carbides and fluorides, and systematically summarized on the basis of their synthesis approaches and mechanisms, as well as properties. Two synthesis routes such as hard-templating (nanocasting) and soft-templating (surfactant assembly) routes are demonstrated. The principal issues in the nanocasting synthesis including the template composition and mesostructure, pore surface chemistry, precursor selection, processing and template removal are emphatically described. A great number of successful cases from the soft-templating method are focused on the surfactant liquid-crystal mesophases to synthesize mesostructured metal chalcogenide composites and the inorganic-block-organic copolymer self-assembly to obtain non-oxide ceramics (296 references). 相似文献
12.
Yu Wen Facai Wei Wenqian Zhang Anyang Cui Jing Cui Chengbin Jing Zhigao Hu Qingguo He Jianwei Fu Shaohua Liu Jiangong Cheng 《中国化学快报》2020,31(2):521-524
Two-dimensional mesoporous materials combing ultrathin nanosheet morphology with well-defined mesoporous structures,are now emerging and becoming increasingly important for their promising applications in energy storage,electronic devices,electrocatalysts and so on.Here,we synthesized a kind of polypyrrole-based two-dimensional mesoporous materials with uniform pore size,ultrathin thickness and high surface area.Serving for electrochemical NH3 sensor,they exhibited a fast response and high sensitivity.Therefore,our study would promote much interest in design of new materials for gas sensor applications. 相似文献
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Rao KS Rani SU Charyulu DK Kumar KN Lee BK Lee HY Kawai T 《Analytica chimica acta》2006,576(2):177-183
A novel approach for immobilization of probe oligonucleotides that uses zirconium phosphate modified silica nanoparticles is proposed. The surface modification of nanoparticles was carried out in two stages. Initially binding of Zr4+ to the surface of silica nanoparticles and later treated with phosphoric acid for terminal phosphate groups. Oligonucleotide probes modified with amine group at 5'-end were strongly binds to the phosphate terminated silica nanoparticles with imidazole in presence of 0.1 mol L(-1) EDC [N-ethyl-N'-(3-dimethylaminopropyl) carbodiimide], as phosphate groups are more reactive towards amine group. Various studies, i.e., synthesis of silica nanoparticles, their surface modification, probe immobilization, measurement of hybridization and effect of bovine serum albumin (BSA) were carried out during optimization of reaction conditions. The significant reduction in the background signal was observed by treating the probe modified silica nanoparticles with bovine serum albumin prior to hybridization. The probe modified silica nanoparticles were retained their properties and the hybridization was induced by exposure of single-stranded DNA (ssDNA) containing silica nanoparticles to the complementary DNA in solution. The decrease in the fluorescence signal for one mismatch and three mismatch was observed upon hybridization of probe with target DNAs, while there was no response for the random target ssDNA under the same experimental conditions. The intensity of fluorescence signal was linear to the concentration of target DNA ranging from 3.9 x 10(-9) to 3.0 x 10(-6)mol L(-1). A detection limit of 1.22 x 10(-9) mol L(-1) of oligonucleotides can be estimated. The proposed hybridization assay is simple and possesses good analytical characteristics and it can provide an effective and efficient route in the development of DNA biosensors and biochips. 相似文献
15.
Summary When mammalian plasma was passed through a chromatographic material containing aminoethyl functional groups, ceruloplasmin was selectively retained. At a specific ionic strength of the eluant buffer, a single chromatographic peak corresponding to the electrophoretically homogeneous purified ceruloplasmin was eluted. This single-step procedure is easy to perform and gave a purification yield of more than 60%. The direct immobilization of the ceruloplasmin, while it was still adsorbed and concentrated at the basal part of the gel bed (last stage of the purification), was achieved by carbodiimide treatment, with coupling yields of 50–70%.The immobilized ceruloplasmin retained about 100% of its enzymatic activity. Kinetic studies have shown a decreased affinity of the immobilized protein for the substrate and a maximal velocity of 81% as compared to the free protein. The immobilized ceruloplasmin was much more resistant to proteolytic attack than the free enzyme which is highly protease sensitive. Using pronase and thermolysine proteases, the activity of free ceruloplasmin was entirely lost in few hours. However, under similar conditions, the immobilized ceruloplasmin exhibited a high stability, maintaining its integral activity even after 24 hours of proteolytic attack. 相似文献
16.
We have investigated the influence of the ratio between the volume of the hydrophilic head ( V A) and the volume of the hydrophobic part ( V B) of the surfactant on the mesopore ordering. To understand the difference of behavior we have performed a complete study dealing with fluorinated [R m (F)(EO) n ] and hydrogenated [R m (H)(EO) n ] surfactants. Their mixtures have also been taken into account. Here only the phase diagrams and the structural parameters of the liquid crystal phases of the mixed systems are reported. We have shown that the mutual or partial miscibility of the fluorinated and the hydrogenated surfactants depends on the number of oxyethylene units of each surfactant. To follow, various systems were used for the preparation of silica mesoporous materials via a cooperative templating mechanism (CTM). Results clearly reveal that V A/ V B ratios in the range between 0.95 and 1.78 lead to the formation of well-ordered mesostructures. Wormhole-like structures are obtained for higher or lower values. Moreover, results show that from the V A/ V B point of view, polyoxyethylene fluoroalkyl ether surfactants behave like their hydrogenated analogues. 相似文献
17.
In this study heparin was covalently immobilized onto LDPE-VEMAC sheet fabricated by the introduction of carboxyl groups to the surface of low-density polyethylene (LDPE) using a plasma technique. The plasma irradiation time influenced the density of carboxyl groups on the LDPE-VEMAC sheet. Heparin was immobilized on the LDPE-VEMAC sheet using a condensation reagent, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC). We confirmed the immobilization of heparin from the ATR-FT-IR spectrum of the sheet obtained. Where heparin was directly immobilized on the LDPE-VEMAC sheet, the density of the immobilized heparin depended on that of the carboxyl groups. Heparin was also immobilized with a spacer, hexamethylene diamine, and the density of such heparin was about 1.6 times that of the directly immobilized heparin. This result suggests that the introduction of a spacer may be an effective way to increase the density of immobilized heparin. 相似文献
18.
Atsuko Yoshida Norishige Kakegawa Makoto Ogawa 《Research on Chemical Intermediates》2003,29(7-9):721-731
The adsorption of a cationic porphyrin, tetrakis-(N-methyl-4-pyridiniumyl)porphine, into mesoporous silicas from solution of tetrakis-(N-methyl-4-pyridiniumyl)porphine p-toluene sulfonate was investigated. Irrespective of the pore size (2.4, 3.5 and 4.2 nm), the cationic porphyrin was adsorbed effectively onto mesoporous silicas to give brown-colored powders. Depending on the amounts adsorbed, which correlate with the average intermolecular distance, the porphines tend to aggregate (dimer). 相似文献
19.
B. SzajÀNi P. SÜdi Gabriella KlamÀr Zsuzsa M. JÀszay I. PetnehÀzy L. TŐke 《Applied biochemistry and biotechnology》1991,30(2):225-231
A series of water-soluble disubstituted carbodiimides of different structure was tested for enzyme immobilization. In the experiments, a polyacrylamide-type bead polymer possessing carboxylic functional groups was used as support. The enzymes immobilized were aminoacylase (N-acylamino acid amidohydrolase; EC 3.5.1.14), arginase (L-arginine amidinohydrolase; EC 3.5.3.1), cyclodextrin glycosyltransferase (alpha-1,4-glucan 4-glycosyltransferase, cyclizing; EC 3.2.1.19), glucoamylase (1,4-alpha-D-glucan glycohydrolase, EC 3.2.1.3), and carboxypeptidase B (peptidyl-L-lysine [L-arginine] hydrolase; EC 3.4.17.2). It was found that the degree of immobilization strongly depended on the structure of carbodiimide used. 相似文献
20.
A. V. Chuenko T. I. Davidenko L. N. Shinkarenko N. R. Suslova V. V. Bolokhovskii 《Chemistry of Natural Compounds》1990,25(4):447-450
The electrooxidation of the alkaloid ephedrine at solid electrodes in a wide range of concentrations and pH values of solutions has been studied by the methods of potentiodynamic voltammetric curves, preparative electrolysis, and quantum-chemical calculations by the SCF MO LCAO method in the MINDO/3 approximation. A quantitative basis has been given for the sequence of stages in the electrooxidation of ephedrine proposed previously by the authors of one of the cited papers. Good agreement has been established between the values found theoretically and experimentally.Institute of Organic Synthesis and Colloid Chemistry, Kazakh SSR Academy of Sciences, Karaganda. Translated from Khimiya Prirodnykh Soedinenii, No. 4, pp. 523–526, July–August, 1989. 相似文献