Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

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)
Please use this url to cite or link to this publication:
author
; ; ; ; and
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
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004), Muscle biology (013212015)
id
dae8dc4d-6138-411d-8c3e-ef3d216e44b6 (old id 138457)
date added to LUP
2016-04-01 15:54:47
date last changed
2022-03-14 20:48:05
@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}},
  doi          = {{10.1016/j.aca.2004.05.048}},
  volume       = {{518}},
  year         = {{2004}},
}