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Specific detection of L-glutamate in food using flow-injection analysis and enzymatic recycling of substrate

Khampha, Wanida LU ; Yakovleva, Julia LU ; Isarangkul, D; Wiyakrutta, S; Meevootisom, V and Emnéus, Jenny LU (2004) In Analytica Chimica Acta 518(1-2). p.127-135
Abstract
A flow injection analysis (FIA) system for specific determination of L-glutamate in food samples based on a bi-enzymatic amplification system has been developed. The content of L-glutamate in the sample was amplified by cycling between L-glutamate dehydrogenase (GIDH) and a novel enzyme, D-phenylglycine aminotransferase (D-PhgAT). In this system, GIDH converts L-glutamate to 2-oxoglutarate with concomitant reduction of NAD(+) to NADH. D-PhgAT transfers an amino group from D-4-hydroxyphenylglycine to 2-oxoglutarate, thus recycling L-glutamate. Accumulation of NADH in the course of the enzymatic recycling was monitored both by fluorescence and UV absorbance and used for quantification of L-glutamate. The assay was characterized by high... (More)
A flow injection analysis (FIA) system for specific determination of L-glutamate in food samples based on a bi-enzymatic amplification system has been developed. The content of L-glutamate in the sample was amplified by cycling between L-glutamate dehydrogenase (GIDH) and a novel enzyme, D-phenylglycine aminotransferase (D-PhgAT). In this system, GIDH converts L-glutamate to 2-oxoglutarate with concomitant reduction of NAD(+) to NADH. D-PhgAT transfers an amino group from D-4-hydroxyphenylglycine to 2-oxoglutarate, thus recycling L-glutamate. Accumulation of NADH in the course of the enzymatic recycling was monitored both by fluorescence and UV absorbance and used for quantification of L-glutamate. The assay was characterized by high long-term stability (at least 70 days) and good reproducibility (within-day and between-day RSDs were 4.3-7.3% and 8.9%). The fluorimetric assay was slightly more sensitive with a L-glutamate detection limit of 0.4 muM and linear range of 2.5-50 muM. The assay was specific for L-glutamate, with recoveries between 95-103% in the presence of 17 different amino acids tested one by one. The method was applied to analysis of real food samples and results were correlated with a commercial Boehringer Mannheim assay kit. (C) 2004 Elsevier B.V. All rights reserved. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytica Chimica Acta
volume
518
issue
1-2
pages
127 - 135
publisher
Elsevier
external identifiers
  • wos:000223005700015
  • scopus:3142674988
ISSN
1873-4324
DOI
10.1016/j.aca.2004.05.048
language
English
LU publication?
yes
id
dae8dc4d-6138-411d-8c3e-ef3d216e44b6 (old id 138457)
date added to LUP
2007-06-27 09:28:27
date last changed
2017-01-01 06:50:15
@article{dae8dc4d-6138-411d-8c3e-ef3d216e44b6,
  abstract     = {A flow injection analysis (FIA) system for specific determination of L-glutamate in food samples based on a bi-enzymatic amplification system has been developed. The content of L-glutamate in the sample was amplified by cycling between L-glutamate dehydrogenase (GIDH) and a novel enzyme, D-phenylglycine aminotransferase (D-PhgAT). In this system, GIDH converts L-glutamate to 2-oxoglutarate with concomitant reduction of NAD(+) to NADH. D-PhgAT transfers an amino group from D-4-hydroxyphenylglycine to 2-oxoglutarate, thus recycling L-glutamate. Accumulation of NADH in the course of the enzymatic recycling was monitored both by fluorescence and UV absorbance and used for quantification of L-glutamate. The assay was characterized by high long-term stability (at least 70 days) and good reproducibility (within-day and between-day RSDs were 4.3-7.3% and 8.9%). The fluorimetric assay was slightly more sensitive with a L-glutamate detection limit of 0.4 muM and linear range of 2.5-50 muM. The assay was specific for L-glutamate, with recoveries between 95-103% in the presence of 17 different amino acids tested one by one. The method was applied to analysis of real food samples and results were correlated with a commercial Boehringer Mannheim assay kit. (C) 2004 Elsevier B.V. All rights reserved.},
  author       = {Khampha, Wanida and Yakovleva, Julia and Isarangkul, D and Wiyakrutta, S and Meevootisom, V and Emnéus, Jenny},
  issn         = {1873-4324},
  language     = {eng},
  number       = {1-2},
  pages        = {127--135},
  publisher    = {Elsevier},
  series       = {Analytica Chimica Acta},
  title        = {Specific detection of L-glutamate in food using flow-injection analysis and enzymatic recycling of substrate},
  url          = {http://dx.doi.org/10.1016/j.aca.2004.05.048},
  volume       = {518},
  year         = {2004},
}