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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Monatshefte für Chemie 115 (1984), S. 1255-1267 
    ISSN: 1434-4475
    Keywords: Enzyme characterisation ; Naringinase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Description / Table of Contents: Zusammenfassung Naringinase ausAspergillus niger zeigt α-L-Rhamnosidase- und β-D-Glucosidaseaktivität. Das Verhältnis von Rhamnosidase- und Glucosidaseaktivität im Enzymkomplex kann sich verändern und hängt von der Proteinkonzentration und dempH ab. Die Rhamnosidaseaktivität des Enzymkomplexes zwischenpH3 undpH7 ist nahezu konstant, während die Glucosidase ein deutliches Optimum besitzt, das allerdings je nach Vorbehandlung zwischenpH4 undpH6 schwanken kann. Durch Gelfiltration kann der Enzymkomplex in verschiedene Oligomere aufgetrennt werden. Alle Fraktionen sind Vielfache der kleinsten aktiven Einheit (M=95 000) und besitzen entweder beide Enzymaktivitäten oder nur Rhamnosidaseaktivität. Aktive Glucosidase allein konnte nicht isoliert werden. Durch Immobilisierung kann jedoch in Proteinfraktionen mit reiner Rhamnosidaseaktivität wieder totale Naringinaseaktivität induziert werden. Die Glukosidaseaktivität ist abhängig von der Proteinkonzentration und verschwindet in sehr verdünnten Lösungen, während die Rhamnosidase unter diesen Bedingungen aktiv bleibt.
    Notes: Abstract Naringinase fromAspergillus niger shows α-L-rhamnosidase and β-D-glucosidase activity. The ratio of these enzymatic activities varies, depending on the protein concentration as well as thepH. The rhamnosidase activity is nearly independent of thepH in the range from 3 to 7, whereas glucosidase shows a distinct optimum which varies betweenpH4 andpH6 depending on its pretreatment. By gel filtration the enzyme complex can be separated into various oligomers, which are multiples of the smallest active subunit with a molecular weight of 95 000. The oligomers show either both enzymatic activities or mere rhamnosidase action. Protein fractions with glucosidase activity only could not be isolated. However in fractions with rhamnosidase activity only, the glucosidase activity could be restored by immobilisation. Glucosidase activity is related to the concentration of protein in solution, disappearing in very diluted solutions, where rhamnosidase is still active.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Monatshefte für Chemie 129 (1998), S. 1067-1092 
    ISSN: 1434-4475
    Keywords: Keywords. Metal nano clusters; Surface enhanced absorption; Array; Teststick.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Description / Table of Contents: Zusammenfassung.  Metallcluster, die durch elektromagnetische Strahlung angeregt werden, zeigen starke lokale Feldüberhöhung und resonantes Verhalten. Absorptive Eigenschaften dieser Metallcluster, welche an oder nahe einer Oberfläche gebunden sind, können als Basis neuartiger analytischer Meßsysteme dienen, welche biorekognitive Wechselwirkungen in ein optisches Signal umzuwandeln vermögen. Immunogoldmarkierung, Immunogoldchromatographie und die SPR-Transduktion der biorekognitiven Bindung von Metallclustern an Oberflächen wurden bereits mit Erfolg kommerziell verwertet. Mehrschichtige hochresonante Systeme wurden zuerst theoretisch berechnet und in den letzten Jahren durch einen Aufbau aus metallischer Spiegelschicht, einer nanometerdicken Polymerschicht, einer biorekognitiven Interaktionsschicht und daran gebundenen metallischen Clustern realisiert. Experimentell konnte gezeigt werden, daß die Symmetrie und der schalenförmige Aufbau der Cluster einen starken Einfluß auf die Lage und das Auftreten von resonanten Absorptionsbanden hat. Insbesondere Cluster mit glasartig-amorpher Struktur zeigen ein schmalba ndiges Reflexionsminimum im Rot- bzw. IR-Bereich und liegen daher weit niederenergetischer als sphärische Goldkolloide (〈 520 nm). Modifizierte Metallcluster, welche durch Unterbrechen des thermischen Reduktionsschritts aus Au3+-Lösungen oder durch Metall-Dielektrikum-Schalenaufbau synthetisch zugänglich sind, zeigen die erwähnte starke Resonanz im IR-Bereich. Der Sensoraufbau ermöglicht es, direkt (d.h. ohne zusätzliche Verfahrensschritte) biorekognitive Bindungsvorgänge und katalytisch-enzymatische Prozesse unter Anwendung der oberflächenverstärkten Clusterabsorption in ein optisches Signal (Farbe der Chipoberfläche) umzuwandeln. Sowohl einschrittige Assays in Form von Teststreifen oder Sensoren als auch kontinuierliche Meßsysteme konnten unter Nutzung von Lectin-Zucker-, Antigen-Antikörper-, Protein-Rezeptor-, Metall-Ligand-oder Nukleinsäurewechselwirkungen aufgebaut werden.
    Notes: Summary.  Metal clusters excited by electromagneti c radiation exhibit high local field enhancement and nanoscale resonant behavior. Absorptive properties of these metal clusters bound to a surface are the basis of various new and highly promising setups to transduce biorecognitive interactions into an optical signal. Immunogold labelling and immunogold chromatography as well as SPR transduction of metal cluster binding have been successfully commercialized. Multilayered highly resonant systems have been proposed and recently realized employing a metal mirror, a polymer distance layer, a biomolecule interaction layer, and biorecognitively bound metal nano clusters. Experiments indicate a strong influence of cluster symmetry and cluster shell on the strong reflection minima induced by the resonant behavior. Modified clusters, clearly exhibit at least one additional narrow reflection minimum in the red or infrared region and therefore far away from spherical gold colloids (〈520 nm). Glass-type metal clusters synthesized by interruption of the thermal step of Au3+-reduction as well as metal-dielectric shell clusters synthesized by multiple shell deposition processes enables the shift to a near-mid IR resonance. It is possible to convert directly (without use of additional analysis steps) biorecognitive binding processes and catalytic activity of proteins by the application of surface enhanced clusters into a visible optical signal (color change of sensor surface). Disposable single step assays as well as multiuse-monitoring devices have been established employing e.g. lectin-sugar, antigen-antibody, protein-receptor, or DNA-DNA interactions.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Microchimica acta 121 (1995), S. 87-93 
    ISSN: 1436-5073
    Keywords: biosensor ; fibre-optic ; mitoxantrone ; absorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract A major problem in therapeutic use of the cytostatic agent mitoxantrone is the occurrence of side effects. Estimation of the drug concentration in or next to the tumor can lead to a reduction of the toxicity through accurate dose adjustment. In this paper a method for the determination of mitoxantrone by a fibre-optic device is described. The advantage of the system is the possibility of remote sensing at the working site of the cytostatic agent by a method that is simple in handling and sufficient in sensitivity. The measuring principle is based on the blue colour of mitoxantrone. The function of the device was tested byin vitro assays; linear calibration curves in a concentration range between 2 and 10μg of mitoxantrone per ml of sample solution were obtained.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Chicester [u.a.] : Wiley-Blackwell
    Journal of Molecular Recognition 9 (1996), S. 644-651 
    ISSN: 0952-3499
    Keywords: lipase ; N-hydroxysuccinimide ; polyethylene glycol ; modification ; carboxylic groups ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Lack of toxicity, excellent solubility and superb biocompatibility make polyethylene glycol (PEG) one of the most popular modifiers of biologicals. The most common method for attachment of PEG is based on modification of amino groups of proteins with methoxy- or succinimide-derivatives of PEG. In the case of proteins with amino groups important for biological activity, this modification can lead to inactivation of proteins. A new strategy for covalent attachment of PEG to carboxylic groups of proteins using O,O-bis-(2-aminopropyl)polyethylene glycol and carbodiimide/N-hydroxysuccinimide-mediated reaction was developed. The reaction is carried out under mild aqueous conditions. The attached PEG serves as a hydrophilic spacer for further bioconjungation with biomolecules and haptens. Lipase from Candida rugosa was used as a model protein. Characteristic data of the modified protein such as activity, isoelectric points and stability were compared with that of the unmodified protein.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Chicester [u.a.] : Wiley-Blackwell
    Journal of Molecular Recognition 8 (1995), S. 139-145 
    ISSN: 0952-3499
    Keywords: lipase ; bio-assay ; biosensor ; hybridisation ; Candida ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Radiolabelled polynucleotide probes have been employed extensively for the detection of complementary nucleic acids by specific hybridization. Within the last few years, various methods have been developed using enzyme-labelled probes to avoid unstable and hazardous isotopes. These assays, based on photometry, fluorescenec, and chemiluminescence, have helped to overcome the use of radioactive probes.To increase the performance of a non-radioactive DNA detection system, the labelling enzyme should remain stable under hybridization conditions which allow the formation of a 15-25 bp long DNA-DNA duplex (Tm = 50-70°C). Therefore, the use of unstable phosphatase and peroxidase conjugates must be avoided due to the composition of the hybridization mixture and the high temperature. By screening various hydrolytic enzymes to fit the special demands, fungal lipases turned out to be the most practical. They offer high sensitivity due to an extremely high turnover number, stability at room temperature for several years, thermostability under working conditions and an easy design of various chromogenic, fluorescent and electrochemical active substrates.Several types of silanized, oxidized and unmodified metal sensors and also standard microtitre plates modified with amino groups were used for the immobilization of oligonuleotides. A sandwich hybridization using the lipase-labelled oligonucleotide probe and a terminal immobilized capture DNA on a microtitre plate or sensor surface combined with a rapid hybridization in solution simplifies and improves the performance of the DNA detection system.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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