Dalwadi, MP;
Orol, D;
Walter, F;
Minton, NP;
King, JR;
Kovacs, K;
(2020)
Using singular perturbation theory to determine kinetic parameters in a non-standard coupled enzyme assay.
Journal of Mathematical Biology
, 81
(2)
pp. 649-690.
10.1007/s00285-020-01524-8.
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Abstract
We investigate how to characterize the kinetic parameters of an aminotransaminase using a non-standard coupled (or auxiliary) enzyme assay, where the peculiarity arises for two reasons. First, one of the products of the auxiliary enzyme is a substrate for the primary enzyme and, second, we explicitly account for the reversibility of the auxiliary enzyme reaction. Using singular perturbation theory, we characterize the two distinguished asymptotic limits in terms of the strength of the reverse reaction, which allows us to determine how to deduce the kinetic parameters of the primary enzyme for a characterized auxiliary enzyme. This establishes a parameter-estimation algorithm that is applicable more generally to similar reaction networks. We demonstrate the applicability of our theory by performing enzyme assays to characterize a novel putative aminotransaminase enzyme, CnAptA (UniProtKB Q0KEZ8) from Cupriavidus necator H16, for two different omega-amino acid substrates.
| Type: | Article |
|---|---|
| Title: | Using singular perturbation theory to determine kinetic parameters in a non-standard coupled enzyme assay |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1007/s00285-020-01524-8 |
| Publisher version: | https://doi.org/10.1007/s00285-020-01524-8 |
| Language: | English |
| Additional information: | © 2021 Springer Nature Switzerland AG. This article is licensed under a Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/). |
| Keywords: | Asymptotic analysis, Reaction kinetics, Synthetic biology, Enzyme characterization |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > UCL BEAMS UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Maths and Physical Sciences > Dept of Mathematics |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10135052 |
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