Complex Media and Enzymatic Kinetics

Evangelos Bakalis, Alice Soldà, Marios Kosmas, Stefania Rapino, Francesco Zerbetto

Research output: Contribution to journalArticle

Abstract

Enzymatic reactions in complex environments often take place with concentrations of enzyme comparable to that of substrate molecules. Two such cases occur when an enzyme is used to detect low concentrations of substrate/analyte or inside a living cell. Such concentrations do not agree with standard in vitro conditions, aimed at satisfying one of the founding hypotheses of the Michaelis-Menten reaction scheme, MM. It would be desirable to generalize the classical approach and show its applicability to complex systems. A permeable micrometrically structured hydrogel matrix was fabricated by protein cross-linking. Glucose oxidase enzyme (GOx) was embedded in the matrix and used as a prototypical system. The concentration of H2O2 was monitored in time and fitted by an accurate solution of the enzymatic kinetic scheme, which is expressed in terms of simple functions. The approach can also find applications in digital microfluidics and in systems biology where the kinetics response in the linear regimes often employed must be replaced.

Original languageEnglish
Pages (from-to)5790-5796
Number of pages7
JournalAnalytical Chemistry
Volume88
Issue number11
DOIs
Publication statusPublished - Jun 7 2016

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Kinetics
Enzymes
Digital microfluidics
Glucose Oxidase
Hydrogel
Substrates
Large scale systems
Cells
Molecules
Proteins
Systems Biology

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Bakalis, E., Soldà, A., Kosmas, M., Rapino, S., & Zerbetto, F. (2016). Complex Media and Enzymatic Kinetics. Analytical Chemistry, 88(11), 5790-5796. https://doi.org/10.1021/acs.analchem.6b00343

Complex Media and Enzymatic Kinetics. / Bakalis, Evangelos; Soldà, Alice; Kosmas, Marios; Rapino, Stefania; Zerbetto, Francesco.

In: Analytical Chemistry, Vol. 88, No. 11, 07.06.2016, p. 5790-5796.

Research output: Contribution to journalArticle

Bakalis, E, Soldà, A, Kosmas, M, Rapino, S & Zerbetto, F 2016, 'Complex Media and Enzymatic Kinetics', Analytical Chemistry, vol. 88, no. 11, pp. 5790-5796. https://doi.org/10.1021/acs.analchem.6b00343
Bakalis E, Soldà A, Kosmas M, Rapino S, Zerbetto F. Complex Media and Enzymatic Kinetics. Analytical Chemistry. 2016 Jun 7;88(11):5790-5796. https://doi.org/10.1021/acs.analchem.6b00343
Bakalis, Evangelos ; Soldà, Alice ; Kosmas, Marios ; Rapino, Stefania ; Zerbetto, Francesco. / Complex Media and Enzymatic Kinetics. In: Analytical Chemistry. 2016 ; Vol. 88, No. 11. pp. 5790-5796.
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