Model-based experimental analysis of enzyme kinetics in aqueous–organic biphasic systems

Zavrel, Michael; Schmidt, Timothy R.; Michalik, Claas; Schwendt, Tilman; Spieß, Antje C.; Ansorge‐Schumacher, Marion B.; Janzen, Christoph; Marquardt, Wolfgang; Büchs, Jochen

doi:10.1016/j.jbiotec.2007.07.148

Cited by 2 publications

(10 citation statements)

References 227 publications

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“…These weighing differences can translate into error propagation at later time points. 19 This observation became apparent in an earlier study for the transesterification of ethyl butyrate over the same temperature range. 17 Presumably higher reaction rates at higher temperatures are overcompensated by increasing destabilization of the enzyme.…”

Section: Resultsmentioning

confidence: 76%

“…Only the last data point obtained after 1440 min shows a discrepancy, which probably arises from slightly different initial compositions. These weighing differences can translate into error propagation at later time points . This observation became apparent in an earlier study for the transesterification of ethyl butyrate over the same temperature range .…”

Section: Kinetics Investigationmentioning

confidence: 82%

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Model validation for enzymatic reactive distillation to produce chiral compounds

Wierschem

Langen

Lins

et al. 2017

J of Chemical Tech & Biotech

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BACKGROUND Enzymatic reactive distillation (ERD) is a biocatalyzed process, in which enzymes are immobilized in catalytic packing. The combination of enzymatic reaction and thermal separation helps to overcome chemical reaction and phase equilibrium limitations. Processing of chiral molecules, in particular, can benefit from ERD application, which might lead to more eco‐efficient processing of these valuable chemicals. Therefore an integrated approach to evaluate this technology is followed. RESULTS To evaluate ERD the transesterification of racemic (R/S)‐1‐phenylethanol (RPE/SPE) to (R)‐phenylethyl acetate (PEA) catalyzed by Candida antarctica lipase B (EC: 3.1.1.3) is investigated with regard to kinetics and physical property data, which provide a basis for the modeling of an ERD process. Furthermore, ERD experiments show a selective conversion of RPE to PEA, which is predicted by the established ERD model with high accuracy. CONCLUSION The ERD experiments demonstrate the feasibility of chiral processing for the transesterification by means of ERD and a validated ERD model is developed, allowing for a conceptual evaluation of ERD. This provides the basis for a future comparison of ERD with benchmark processes that will reveal the economic potential of ERD processes. © 2017 Society of Chemical Industry

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Section: Resultsmentioning

confidence: 76%

Section: Kinetics Investigationmentioning

confidence: 82%

Model validation for enzymatic reactive distillation to produce chiral compounds

Wierschem

Langen

Lins

et al. 2017

J of Chemical Tech & Biotech

View full text Add to dashboard Cite

show abstract

“…[4][5][6] It was shown that the classical approach of taking samples from the organic phase is insufficient to correctly understand the processes. 7 Therefore, improved measurement techniques are necessary and will be introduced in detail below. It was demonstrated that measurements in the bead center are well suited for the identification of the diffusion kinetics.…”

Section: Introductionmentioning

confidence: 99%

“…8 On the other hand, it was shown that gathering measurement information along the radius of a bead is optimal in the case of a reactive hydrogel system. 7 For diffusion measurements in hydrogels, established methods are Fourier transform infrared spectroscopy (FT-IR) and, in the case of non-fluorescent reactants, Raman spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. [9][10][11] Confocal laser scanning microscopy (CLSM) is used for qualitative analysis of protein distribution in hydrogel beads.…”

Section: Introductionmentioning

confidence: 99%

“…Afterwards, the raw data is used for parameter estimation and determination of kinetic constants in order to establish a mechanistic kinetic model that takes all relevant phenomena into account. 7 The fluorescence of the substrate enables the direct quantification of concentration gradients using a multiphoton approach. In this work, new techniques of microscopic resolution are demonstrated for the noninvasive detection of concentration gradients in enzyme-catalyzed biphasic spatially distributed systems.…”

Section: Introductionmentioning

confidence: 99%

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Determination of Temporal and Spatial Concentration Gradients in Hydrogel Beads Using Multiphoton Microscopy Techniques

Schwendt

Michalik²,

Zavrel³

et al. 2010

Appl Spectrosc

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Multiphoton microscopy is a promising technique to detect spatially and temporally resolved concentration gradients of chemical compounds, e.g., reactants in hydrogel-encapsulated biocatalysts. In contrast to current techniques, the improved spatial and temporal resolution of this method in data acquisition and its ability to measure hydrogel beads facilitates the identification of various kinetic phenomena. To our knowledge, multiphoton microscopy is used here for the first time to examine diffusion, mass transfer, and reaction in immobilized hydrogel systems. In a first step, the phenomena of diffusion and diffusion-coupled mass transfer through the phase interface are investigated in the bead center. Finally, the complete system--consisting of diffusion, mass transfer, and enzymatic reaction--is observed by measuring concentration gradients along the bead radius with temporal and spatial resolution. This metrology enables a subsequent mechanistic model identification, which in turn leads to an enhanced knowledge of reaction kinetics and supports the design of biotechnological processes. This task was only possible due to excellent spatial (25 microm) and temporal (5 s) resolution and the accuracy (+/-1%) achieved by using a multiphoton microscopy setup.

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Model-based experimental analysis of enzyme kinetics in aqueous–organic biphasic systems

Cited by 2 publications

References 227 publications

Model validation for enzymatic reactive distillation to produce chiral compounds

Model validation for enzymatic reactive distillation to produce chiral compounds

Determination of Temporal and Spatial Concentration Gradients in Hydrogel Beads Using Multiphoton Microscopy Techniques

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