20.500.12556/RUL-182932 Addressing sensitivity and non-uniqueness in the determination of enzyme kinetic parameters research data underlying the article Accurate determination of enzyme kinetic parameters is critical for model-based design and intensification of biocatalytic processes, particularly in microscale systems. While Michaelis-Menten kinetics provides a foundational framework, its extension to reversible, multi-substrate, and inhibited reactions introduces significant challenges in parameter estimation-most notably, parameter sensitivity and non-uniqueness. This study systematically investigates these challenges across three case studies of increasing complexity: (i) mono-substrate Michaelis-Menten kinetics, (ii) reversible enzymatic reactions with four parameters, and (iii) a six-parameter reversible mono-substrate kinetic model with substrate and product inhibition. In the first two cases, we show that vastly different parameter sets can yield nearly indistinguishable model fits to experimental data, exposing the limitations of classical graphical and nonlinear regression methods. In the mono-substrate case based on real experimental data, two parameter sets differing by nearly two orders of magnitude produce virtually identical model outputs, demonstrating practical non-uniqueness even for simple kinetic models. For the six-parameter inhibited system, a theoretical and numerical analysis reveals intrinsic non-uniqueness of the parameter estimation problem, characterized by an infinite family of parameter vectors yielding identical solutions. These results demonstrate that parameter non-uniqueness is not merely a consequence of experimental noise, but a structural property of complex kinetic models, emphasizing the need for more robust and structurally informed modeling approaches in biocatalysis. enzyme kinetics kinetic parameter estimation parameter non-uniqueness encimska kinetika določevanje kinetičnih parametrov neenoličnost parametrov true false false Angleški jezik Slovenski jezik Neznano 2026-05-28 21:56:09 2026-05-28 22:21:14 2026-05-29 11:03:42 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 README.pdf README.pdf README 1 397F43B62E82BCE52E6A8151B88C9E4A d6ac1aae92cf5a2a64d598fcc2352ea45574a30eb2e1b5b8dfced04ab22087db 66902b31-5ad1-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=234621 QABEQ.nb QABEQ.nb QABEQ 1 E4CAB18B77FA4A73E3E14F38642ABB01 ceed2a9f341438b6355a71088fabd15a89b31a41d0bb3eb3c6de15beb5bc459c 9ad6a760-5ad1-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=234622 QABEQ.pdf QABEQ.pdf QABEQ 1 E72F24229A71205F36AF1D1D95643E4F ec6b04469d444965b9c0c23ab72ce0b4d2c9fd1d9c0d6fe454472151463da2ae ba7eb1cc-5ad1-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=234623 http://silverstripe.fkit.hr/cabeq/past-issues/article/2459 Article 0 4b0e13f6-5ad2-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=234624 Fakulteta za kemijo in kemijsko tehnologijo Meritve in testi 0 0 0