Automated method, based on micro-sequential injection, for the study of enzyme kinetics and inhibition.

A micro-reactor system with continual spectrophotometric detection has been operated in Sequential Injection lab-on-valve (SI-LOV) mode and applied to enzyme kinetics and inhibition studies, using acetylcholinesterase (AChE) and angiotensin-converting enzyme (ACE) as model systems. With the advantages of automation, real-time kinetic measurement, and thorough mixing, the SI-LOV micro-reactor system allows for the monitoring of initial reaction rates and determination of reactant concentrations in the reaction mixture, both of which are essential for the determination of kinetic constants for enzymes and inhibitors. Enzyme, substrate, and inhibitor are precisely metered by the syringe pump and delivered to a stirred micro-reactor, followed by a reference scan that establishes the baseline for the following reaction rate measurement. Michaelis constants (Km) for AChE and ACE were determined to be 0.16 mM and 0.30 mM, respectively, which are consistent with literature values. The type of inhibition (competitive, uncompetitive, or mixed), the dissociation constants for the inhibitors, and the inhibitor dose-response curves were also determined.     

Remarks on dispersionless KP,KdV, and 2D gravity

Summary We use the SDiff(2) framework of Takasaki and Takebe and the (L, M) program (L is the Lax operator andMω=ω_λ) to show that =semiclassical limit ofM is , where ( ) are action angle variables in the Gibbons-Kodama theory of Hamilton-Jacobi type for dispersionless KP. We also show is the semiclassical limit ofWxW ⁻¹ (W is the gauge operator), whereG=WxW ⁻¹ is a quantity studied by the author in an earlier paper in connection with symmetries. We give then a semiclassical version of the Jevicki-Yoneya action principle for 2D gravity, where again arises in calculations, and this yields directly the Landau-Ginsburg equation that corresponds to the semiclassical limit of an integrated string equation. For KdV we also show how inverse scattering data are connected to Hamiltonians for dispersionless KdV. We also discuss Hirota bilinear formulas relative to the dispersionless hierarchies and establish various limiting formulas.     

An exact numerical analysis of coupled planar electron waveguides with asymmetric rectangular potential profiles

An exact numerical analysis is given of electron waves in a pair of coupled semiconductor heterojunction electron waveguides modelled by a rectangular potential distribution. The waveguide lengths required for electron current transfer between the guides agree asymptotically with estimates from Yariv's coupled mode theory and are usefully more accurate. Moreover the electron wave coupling due to an ideal launcher can be worked out, leading to estimates of the off/on current ratio occurring in the waveguides when regarded as an ultrafast switch. For a Poisson probability distribution of the number of electrons N making up a bit-pulse, we have calculated the minimum average N, dependent upon , which would be required to keep the switching bit-error rate below prescribed limits. Pulse rise times and switching delays are derived from the explicit forms for the longitudinal electron wave group velocity. Combining the bit-error and switching-time calculations leads to estimates of the waveguide current densities which would be required in practice. Some normalized sets of curves relating to our analysis are given and used to predict the performance of particular examples of recently proposed AlGaAs/GaAs heterojunction electron waveguide switch/couplers.     

The oxidative decarboxylation of polyaminocarboxylic acids

Summary The rates of oxidation of four chelating agents, NTA, EDTA, CDTA, and DTPA with Ce(IV), in sulfuric acid media, were determined spectrophotometrically by a stopped-flow technique. The reductive ability is in the order CDTA > EDTA > DTPA > NTA. The influence of varying the acidity of the medium was studied, and in each case a maximum in the rate constant vs. [H⁺] plot was observed. A possible interpretation of the reactivities and the influence of acidity is advanced.

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Oxydative Decarboxylierung von PolyaminocarbonsäurenII. Vergleichende kinetische Untersuchung der Oxydation von NTA, ÄDTA, CDTA und DTPA mit Ce(IV) in saurer Lösung

Zusammenfassung Die Oxydationsgeschwindigkeiten von 4 Chelaten (NTA, ÄDTA, CDTA und DTPA) mit Ce(IV) in saurer Lösung wurden spektrophotometrisch mit Hilfe der stopped-flow-Technik bestimmt. Die Reduzierfähigkeit nimmt in der Reihenfolge CDTA > ÄDTA > DTPA > NTA ab. Der Einfluß verschiedener Säuregehalte in der Lösung wurde untersucht, und in jedem Fall wurde ein Maximum in der graphischen Darstellung der Geschwindigkeitskonstante gegen [H⁺] beobachtet. Eine mögliche Erklärung des Reaktionsvermögens und des Säureeinflusses wird gegeben.

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Part I: Z. Anal. Chem. 246, 231 (1969).     

Retaining and recovering enzyme activity during degradation of TCE by methanotrophs

To determine if compounds added during trichloroethylene (TCE) degradation could reduce the loss of enzyme activity or increase enzyme recovery, different compounds serving as energy and carbon sources, pH buffers, or free radical scavengers were tested. Formate and formic acid (reducing power and a carbon source), as well as ascorbic acid and citric acid (free radical scavengers) were added during TCE degradation at a concentration of 2 mM. A saturated solution of calcium carbonate was also tested to address pH concerns. In the presence of formate and methane, only calcium carbonate and formic acid had a beneficial effect on enzyme recovery. The calcium carbonate and formic acid both reduced the loss of enzyme activity and resulted in the highest levels of enzyme activity after recovery.