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MALATE DETERMINATION BY FLOW INJECTION ANALYSIS ( FIA ) USING A COUPLED DEHYDROGENASE/TRANSFERASE SYSTEM
Chemnitius, Gabriele C. Schmid, Rolf D.
Chemnitius, Gabriele C.
Schmid, Rolf D.
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Issue Date
1989
Submitted date
2024-01-16
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Abstract
The determination of L-malate was based on its oxidation by NADt catalyzed by immobilized malate dehydrogenase (MDH, EC 1.1.1.37) in a flow injection system. The NADH produced in an enzyme reactor was detected fluorimetrically. Malate dehydrogenases from different sources were immobilized on aminoderivatized controlled pore glass by glutardialdehyde. Hence the equilibrium of the MDH catalyzed reaction is far in favour of L-malate only little response could be obtained by malate injection. To achieve higher substrate conversion, aspartate aminotransferase (AST, EC 2.6.1.1) was coimmobilized to transform the reaction product oxaloacetate. Enzyme reactors with and without AST were compared and optimized. With the bienzyme reactor, malate concentrations between 5 and 500 uM could be monitored.
Citation
Biosensors : applications in medicine, environmental protection and process control, 299 - 302
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Book chapter
conference paper
conference paper
Language
en
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Series/Report no.
GBF monographs ; Volume 13
ISSN
0930-4320
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ISBN
3527280324
0895739550
0895739550
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Attribution-NonCommercial-ShareAlike 4.0 International