Enzyme merge

The simplest design for an enzyme reactor is to merely have the substrate and enzyme in two merging buffer streams followed by a reaction delay coil (Fig. 12). The delay... 

Fig. 12. On-line enzyme reactor system designs, merging stream system (Top) and immobilized-enzyme reactor system (Bottom). A = mobile phase, B = enzyme solution...

Nakayama K, Puchkaev A, Pikuleva lA. 2001. Membrane binding and substrate access merge in cytochrome P450 7A1, a key enzyme in degradation of cholesterol. J Biol... 

Figure 3.10 — Flow manifolds for implementation of flow-through biosensors. (A) Flow injection merging-zones manifold for the bioluminescence detennination of ATP. ATP standards (30 fiL) and luciferin (30 fiL) are injected into the buffered carrier streams, each pumped at 0.7 mL/min and synchronously merged 12.5 cm downstream. Distance from merging point to immobilized enzyme coil, 2.2 cm. (Reproduced from [59] with permission of Elsevier Science Publishers). (B) Completely continuous flow manifold for the determination of NADH. (Reproduced from [71] with permission of the Royal Society of Chemistry). (C) Segmented-flow manifold for the determination of L-(+)-lactate. (Reproduced from [65] with permission of Marcel Dekker, Inc.). (D) Single-channel flow injection manifold with immobilized reagent for the detennination of glucose. (Reproduced from [77] with permission of Elsevier Science Publishers).

The mixture was finally merged with the bioluminescent solution (FMN + DTT) prior to arrival at the biosensor (a 5-mm x 0.5 mm ID nylon coil containing the luminescent enzymes wound around a Plexiglass support positioned in front of the photomultiplier tube), where the following reactions occurred ... 

The active T-4 circulating in the vascular system merges with receptors and triggers metabolic activity but when it reaches the liver it is changed into the more active thyroid hormone L-Triiodothyronine (T-3) by an enzyme called 5-deiodinase. T-3 is about 5 times more active than T-4. The newly formed T-3 is released into the vascular system where it may contact and merge with cellular receptors which initiates all the metabolic activity discussed earlier. 

M 16] [P 16] The same findings as for the chemical reaction (see Chemical reaction in electrode dot devices with sequential voltage) were made for the luciferin-luciferase enzyme reaction with adenosine triphosphate on the six-phase electrode array device [100, 101]. The reaction rapidly followed the mixing, as evident from the luciferin luminescence of the droplet after merging. 

The amount of immobilized protein can be determined by the mass balance between initial and final solutions [56], A combined qualitative method merged from the classical in-situ detection of enzyme activity and western blot analysis can be applied to determine the enzyme spatial distribution through the membrane thickness and along the membrane module and its activity after the immobilization [57-59]. 

A merging of chemistry and biology is essential to effectively probe the immune system for catalytic antibodies (Fig. 3). Haptens that are successful in eliciting catalytic antibodies are variations of the central theme that transition state stabilization in the antibody combining site will yield functional catalysts for a desired chemical reaction. The evolution of hapten design will be discussed further in subsequent sections. Once the hapten is selected and synthesized, it is attached to an immunogenic carrier protein, usually via an amide bond, for hyperimmunization. A preliminary screen for antibodies that bind the hapten using an enzyme-linked immunosorbent assay (ELISA) is followed by another screen for catalysis of the reaction for which the hapten... 

Restriction maps of entire genomes can be obtained by two approaches. In the top-down approach, infrequently-cutting restriction enzymes and pulsed-field gel electrophoresis are used to map a chromosome much as one would a plasmid. In the bottom-up approach, analysis of many clones in parallel leads to the accumulation of a set of clones which represent the whole genome. Restriction maps of the clones can then be merged to make the genome map. The clones themselves are also a valuable resource. The earliest complete and nearly complete physical mapping projects, both top-down E. co//[10], Schizosaccharomycespombe 19 ) and bottom-up ( . coli[ 1], Saccharomyces cerevisiae[ 2, Caenorhabditis [80,81 ]), were on genomes which were already... 

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