Method for the enzymatic determination of aspartame

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase

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435 23, 435 25, 435 26, 435288, 435808, 435810, C12Q 137

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active

052799450

ABSTRACT:
The aspartame content in aqueous solutions is determined by enzymatic cleavage of aspartame to aspartic acid and phenylalanine methyl ester by means of a peptidase, followed by a detection reaction cocatalyzed by adenine dinucleotide. The phenylalanine is, after elimination of the ester group by means of chymotrypsin, converted by phenylalanine dehydrogenase in the presence of NAD.sup.+ into phenylpyruvate, and the aspartame concentration is measured via the NADH or NH.sub.3 formation which occurs as a consequence. Alternatively, the aspartic acid is converted by a cell extract which acts on aspartic acid in the presence of NADP.sup.+, and the resultant NADPH or NH.sub.3 is used to determine aspartame concentration. These enzymatic reactions preferably are carried out in consecutive enzyme columns with carrier-immobilized enzymes by the FIA technique. The proportion of hydrolysis products can be measured by analyzing another sample omitting the peptidase reaction. The enzymatic cell extract used for the aspartic acid determination is obtained from microorganisms which are isolated from decomposing organic matter. Preferably used for this purpose is the strain DSM 6705. The cell extract can be used as crude extract, without special purification for the analysis, and is generally suitable for detecting aspartic acid.

REFERENCES:
A. Mulchandani et al; Analytica Chimica Acta, 234, (1990) pp. 465-469.
G. G. Guilbault et al; Analytica Chimica Acta, 206, (1988), pp. 369-374.
O. Fatibello-Filho et al; Anal. Chem. 60, (1988), pp. 2397-2399.
P. Schadewaldt et al; Clinica Chimica Acta, 183, (1989) pp. 171-182.
W. Hummel et al; Appl Microbiol Biotechnol, 21 (1985) pp. 7-15.

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