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 language | English |
|---|---|
| Pages (from-to) | 5790-5796 |
| Number of pages | 7 |
| Journal | Analytical Chemistry |
| Volume | 88 |
| Issue number | 11 |
| DOIs | |
| Publication status | Published - Jun 7 2016 |
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ASJC Scopus subject areas
- Analytical Chemistry
Cite this
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 journal › Article
}
TY - JOUR
T1 - Complex Media and Enzymatic Kinetics
AU - Bakalis, Evangelos
AU - Soldà, Alice
AU - Kosmas, Marios
AU - Rapino, Stefania
AU - Zerbetto, Francesco
PY - 2016/6/7
Y1 - 2016/6/7
N2 - 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.
AB - 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.
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U2 - 10.1021/acs.analchem.6b00343
DO - 10.1021/acs.analchem.6b00343
M3 - Article
AN - SCOPUS:84973636784
VL - 88
SP - 5790
EP - 5796
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 11
ER -