A Model System for a Fluorometric Biosensor Using Permeabilized Zymomonas mobilis or Enzymes with Protein Confined Dinucleotides

authored by: O. Thordsen, S. J. Lee, A. Degelau, T. Scheper, H. Loos, B. Rehr, H. Sahm
Abstract: Using permeabilized Zymomonas mobilis or glucose–fructose oxidoreductase isolated from this microorganism a model system for biosensors with a protein confined NADP(H) cofactor for the determination of glucose, fructose, gluconolactone, and sorbitol was developed. Either permeabilized microorganisms containing the oxidoreductase or the pure enzyme were confined via membrane separation in a small measuring chamber, that was integrated into a flow injection analysis system (FIA). The measuring principle was the monitoring of the NAD(P)H fluorescence, excited at 360 nm and measured at 450 nm. NADP(H), which is confined in the protein complex, was oxidized or reduced during the enzymatic reactions and the changes in the fluorescence intensity were related to the substrate concentration. The sensitivity of the system covered a range from 0.001 to 100 g/L of the analyte depending on substrate and operating conditions. The applicability of this model system for bioprocess monitoring was proved using samples from a Pseudomonas pseudoflava cultivation. © 1993 John Wiley & Sons, Inc.
External Organisation(s): University of Münster
Forschungszentrum Jülich
Type: Article
Journal: Biotechnology and Bioengineering
Volume: 42
Pages: 387-393
No. of pages: 7
ISSN: 0006-3592
Publication date: 07.1993
Publication status: Published
Peer reviewed: Yes
ASJC Scopus subject areas: Biotechnology, Bioengineering, Applied Microbiology and Biotechnology
Electronic version(s): https://doi.org/10.1002/bit.260420317 (Access: Unknown)

BibTeX

@article{9553eebe30e049d392df80dfa51db2d7,
title = "A Model System for a Fluorometric Biosensor Using Permeabilized Zymomonas mobilis or Enzymes with Protein Confined Dinucleotides",
abstract = "Using permeabilized Zymomonas mobilis or glucose–fructose oxidoreductase isolated from this microorganism a model system for biosensors with a protein confined NADP(H) cofactor for the determination of glucose, fructose, gluconolactone, and sorbitol was developed. Either permeabilized microorganisms containing the oxidoreductase or the pure enzyme were confined via membrane separation in a small measuring chamber, that was integrated into a flow injection analysis system (FIA). The measuring principle was the monitoring of the NAD(P)H fluorescence, excited at 360 nm and measured at 450 nm. NADP(H), which is confined in the protein complex, was oxidized or reduced during the enzymatic reactions and the changes in the fluorescence intensity were related to the substrate concentration. The sensitivity of the system covered a range from 0.001 to 100 g/L of the analyte depending on substrate and operating conditions. The applicability of this model system for bioprocess monitoring was proved using samples from a Pseudomonas pseudoflava cultivation. {\textcopyright} 1993 John Wiley & Sons, Inc.",
keywords = "bioprocess monitoring, biosensor, fluorescence, fructose, gluconolactone, glucose, NADPH, permeabilization, sorbitol, Zymomonas mobilis",
author = "O. Thordsen and Lee, {S. J.} and A. Degelau and T. Scheper and H. Loos and B. Rehr and H. Sahm",
year = "1993",
month = jul,
doi = "10.1002/bit.260420317",
language = "English",
volume = "42",
pages = "387--393",
journal = "Biotechnology and Bioengineering",
issn = "0006-3592",
publisher = "Wiley-VCH Verlag",
number = "3",
}