A model system for a fluorometric biosensor using permeabilized Zymomonas mobilis or enzymes with protein confined dinucleotides

Journal Article · Thu Jul 01 00:00:00 EDT 1993 · Biotechnology and Bioengineering; (United States)

DOI:https://doi.org/10.1002/bit.260420317· OSTI ID:6289757

Thordsen, O; Lee, S J; Degelau, A ^[1]; Scheper, T ^[2]; Loos, H; Rehr, B; Sahm, H ^[3]

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1. Univ. Hannover (Germany)
2. Univ. Muenster (Germany)
3. Inst. fuer Biotechnologie, Juelich (Germany)

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.

• OSTI ID: 6289757
• Journal Information: Biotechnology and Bioengineering; (United States), Vol. 42:3; ISSN 0006-3592
• Country of Publication: United States
• Language: English

Related Subjects

09 BIOMASS FUELS
OXIDOREDUCTASES
ENZYME ACTIVITY
SORBITOL
BIOSYNTHESIS
ZYMOMONAS MOBILIS
GROWTH
BIOLOGICAL MODELS
FLUORESCENCE
FRUCTOSE
GLUCOSE
NADP
NUCLEOTIDES
PERMEABILITY
REDOX REACTIONS
SENSITIVITY ANALYSIS
ALDEHYDES
BACTERIA
CARBOHYDRATES
CHEMICAL REACTIONS
COENZYMES
DIURETICS
DRUGS
ENZYMES
HEXOSES
KETONES
LUMINESCENCE
MICROORGANISMS
MONOSACCHARIDES
ORGANIC COMPOUNDS
PROTEINS
SACCHARIDES
SYNTHESIS
090900* - Biomass Fuels- Processing- (1990-)