Multiplexed Capillary Electrophoresis as Analytical Tool for Fast Optimization of Multi-Enzyme Cascade Reactions - Synthesis of Nucleotide Sugars

2016 ◽  
Vol 11 (10) ◽  
pp. 1298-1308 ◽  
Author(s):  
Claudia Wahl ◽  
Dennis Hirtz ◽  
Lothar Elling
Keyword(s):  
Capillary Electrophoresis ◽  
Cascade Reactions ◽  
Analytical Tool ◽  
Enzyme Cascade ◽  
Nucleotide Sugars ◽  
Multiplexed Capillary Electrophoresis ◽  
Fast Optimization

scholarly journals Microfluidics Featuring Multilayered Hydrogel Assemblies Enable Real-Time NMR-Monitoring of Enzyme Cascade Reactions

2020 ◽  
Author(s):  
Nurdiana Nordin ◽  
lorenzo bordonali ◽  
Hossein Davoodi ◽  
Novindi Dwi Ratnawati ◽  
Gudrun Gygli ◽  
...  
Keyword(s):  
Real Time ◽  
Functional Performance ◽  
Cascade Reactions ◽  
Inhibitory Effect ◽  
Resonance Spectroscopy ◽  
Reaction Cell ◽  
Enzyme Cascade ◽  
Sensing Applications ◽  
Progress Curve ◽  
Michaelis Constants

Compartmentalized chemical reactions at the microscale are interesting from many perspectives including (multi)functional surfaces and biotechnology. Monitoring the molecular content as a measure of functional performance at these small scales is challenging. As a means to address this challenge, we leverage microtechnology and biocompatible materials to integrate a compact, reconfigurable reaction cell featuring electrochemical functionality with high-resolution nuclear magnetic resonance spectroscopy (NMR). We demonstrate the operation of this system by monitoring the activity of enzymes immobilized in chemically distinct layers within a multi-layered chitosan hydrogel assembly. As a benchmark, we observed the parallel activities of urease (Urs), catalase (Cat), and glucose oxidase (GOx) by recording NMR spectra to extract reagent and product concentrations in real-time. As a result, simultaneous monitoring of a cooperative enzymatic process (GOx + Cat) together with an independent process (Urs) is achieved. Using Michaelis-Menten progress curve analysis of the NMR data, kinetic data is extracted: in the case of GOx, the Michaelis constants (K<sub>M</sub>) are consistent with previous reports, while for Urs, deviations are observed, attributed to an inhibitory effect under our reaction conditions. The system therefore enables the construction of complex reaction cascades with spatial control, as would be interesting in, for example, metabolic engineering and multiplexed sensing applications.

Multi‐enzyme Cascade Reactions in Metal‐organic Frameworks

2020 ◽  
Vol 20 (10) ◽  
pp. 1100-1116 ◽  
Author(s):  
Jieying Liang ◽  
Kang Liang
Keyword(s):  
Metal Organic Frameworks ◽  
Cascade Reactions ◽  
Enzyme Cascade ◽  
Metal Organic

scholarly journals Recent Advances in Enzymatic and Chemoenzymatic Cascade Processes

Catalysts ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 1258
Author(s):  
Noelia Losada-Garcia ◽  
Zaida Cabrera ◽  
Paulina Urrutia ◽  
Carla Garcia-Sanz ◽  
Alicia Andreu ◽  
...  
Keyword(s):  
Organic Chemistry ◽  
Solid Phase ◽  
Cascade Reactions ◽  
Review Article ◽  
The Novel ◽  
Enzyme Cascade ◽  
Cascade Processes ◽  
Novel Strategy ◽  
Substantial Interest

Cascade reactions have been described as efficient and universal tools, and are of substantial interest in synthetic organic chemistry. This review article provides an overview of the novel and recent achievements in enzyme cascade processes catalyzed by multi-enzymatic or chemoenzymatic systems. The examples here selected collect the advances related to the application of the sequential use of enzymes in natural or genetically modified combination; second, the important combination of enzymes and metal complex systems, and finally we described the application of biocatalytic biohybrid systems on in situ catalytic solid-phase as a novel strategy. Examples of efficient and interesting enzymatic catalytic cascade processes in organic chemistry, in the production of important industrial products, such as the designing of novel biosensors or bio-chemocatalytic systems for medicinal chemistry application, are discussed

Enzyme cascade reactions: synthesis of furandicarboxylic acid (FDCA) and carboxylic acids using oxidases in tandem

2015 ◽  
Vol 17 (6) ◽  
pp. 3271-3275 ◽  
Author(s):  
Shane M. McKenna ◽  
Silke Leimkühler ◽  
Susanne Herter ◽  
Nicholas J. Turner ◽  
Andrew J. Carnell
Keyword(s):  
Carboxylic Acids ◽  
Cascade Reactions ◽  
Preparative Scale ◽  
Mild Conditions ◽  
Enzyme Cascade ◽  
Furandicarboxylic Acid

Three enzymes are combined under mild conditions for the preparative scale oxidation of HMF to FDCA and a range of 10 alcohols.

scholarly journals Multi-enzyme cascade reactions using protein–polymer surfactant self-standing films

2017 ◽  
Vol 53 (13) ◽  
pp. 2094-2097 ◽  
Author(s):  
Thomas Farrugia ◽  
Adam W. Perriman ◽  
Kamendra P. Sharma ◽  
Stephen Mann
Keyword(s):  
Cascade Reactions ◽  
Hierarchical Assembly ◽  
Protein Polymer ◽  
Enzyme Cascade ◽  
Catalytically Active ◽  
Polymer Surfactant

Self-supporting bio-catalytically active multi-enzyme films fabricated via hierarchical assembly of enzyme–polymer surfactant nanoconjugates are capable of sustaining cascade reactions.

Evaluation of the Potential of a Charge-Injection Device for DNA Sequencing by Multiplexed Capillary Electrophoresis

1995 ◽  
Vol 49 (6) ◽  
pp. 825-833 ◽  
Author(s):  
Qingbo Li ◽  
Edward S. Yeung
Keyword(s):  
Capillary Electrophoresis ◽  
Dna Sequencing ◽  
Fluorescence Detection ◽  
Charge Injection ◽  
Laser Induced Fluorescence ◽  
Array Detector ◽  
Injection Device ◽  
Laser Induced Fluorescence Detection ◽  
Array Detectors ◽  
Multiplexed Capillary Electrophoresis

Despite the rapid growth in the use of imaging detectors in spectroscopy, the charge-injection device (CID) has unique features that have not been fully exploited. The advantages of the CID as a two-dimensional array detector for laser-induced fluorescence detection in highly multiplexed capillary electrophoresis are evaluated. In such a system, the CID maintains both high sensitivity and high sampling rate, which are usually difficult to achieve simultaneously with other array detectors. Applying the electronic windowing function significantly improves the scan rate and greatly reduces the volume of data generated. With 1-s exposure time and 488-nm excitation, the detection limit of the system is 10−12 M fluorescein with the device cryogenically cooled and 10−11 M fluorescein at ambient temperature. The low dark current of the CID imager allows operation at room temperature without significantly affecting sensitivity when combined with moderate laser powers. We demonstrate that the CID is well suited for high-speed, high-throughput DNA sequencing based on multiplexed capillary electrophoresis with on-column laser-induced fluorescence detection.

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