Specific detection of L-glutamate in food using flow-injection analysis and enzymatic recycling of substrate
(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:
https://lup.lub.lu.se/record/138457
- author
- Khampha, Wanida LU ; Yakovleva, Julia LU ; Isarangkul, D ; Wiyakrutta, S ; Meevootisom, V and Emnéus, Jenny LU
- organization
- publishing date
- 2004
- 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}}, }