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Flavocytochrome b(2)-Based Enzymatic Method of L-Lactate Assay in Food Products
L-lactate, a key metabolite of the anaerobic glycolytic pathway, plays an important role as a biomarker in medicine, in the nutritional sector and food quality control. For these reasons, there is a need for very specific, sensitive, and simple analytical methods for the accurate L-lactate measuring...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi Publishing Corporation
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3800577/ https://www.ncbi.nlm.nih.gov/pubmed/24223505 http://dx.doi.org/10.1155/2013/461284 |
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author | Smutok, Oleh Karkovska, Maria Smutok, Halyna Gonchar, Mykhailo |
author_facet | Smutok, Oleh Karkovska, Maria Smutok, Halyna Gonchar, Mykhailo |
author_sort | Smutok, Oleh |
collection | PubMed |
description | L-lactate, a key metabolite of the anaerobic glycolytic pathway, plays an important role as a biomarker in medicine, in the nutritional sector and food quality control. For these reasons, there is a need for very specific, sensitive, and simple analytical methods for the accurate L-lactate measuring. A new highly selective enzymatic method for L-lactate determination based on the use of flavocytochrome b (2) (EC 1.1.2.3; FC b (2)) isolated from the recombinant strain of the yeast Hansenula polymorpha has been developed. A proposed enzymatic method exploits an enzymatic oxidation of L-lactate to pyruvate coupled with nitrotetrazolium blue (NTZB) reduction to a colored product, formazan. The maximal absorption peak of the colored product is near λ = 525 nm and the linear range is observed in the interval 0.005–0.14 mM of L-lactate. The main advantages of the proposed method when compared to the LDH-based routine approaches are a higher sensitivity (2.0 μM of L-lactate), simple procedure of analysis, usage of inexpensive, nontoxic reagents, and small amount of the enzyme. Enzymatic oxidation of L-lactate catalyzed by flavocytochrome b (2) and coupled with formazan production from nitrotetrazolium blue was shown to be used for L-lactate assay in food samples. A high correlation between results of the proposed method and reference ones proves the possibility to use flavocytochrome b (2)-catalysed reaction for enzymatic measurement of L-lactate in biotechnology and food chemistry. |
format | Online Article Text |
id | pubmed-3800577 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-38005772013-11-10 Flavocytochrome b(2)-Based Enzymatic Method of L-Lactate Assay in Food Products Smutok, Oleh Karkovska, Maria Smutok, Halyna Gonchar, Mykhailo ScientificWorldJournal Research Article L-lactate, a key metabolite of the anaerobic glycolytic pathway, plays an important role as a biomarker in medicine, in the nutritional sector and food quality control. For these reasons, there is a need for very specific, sensitive, and simple analytical methods for the accurate L-lactate measuring. A new highly selective enzymatic method for L-lactate determination based on the use of flavocytochrome b (2) (EC 1.1.2.3; FC b (2)) isolated from the recombinant strain of the yeast Hansenula polymorpha has been developed. A proposed enzymatic method exploits an enzymatic oxidation of L-lactate to pyruvate coupled with nitrotetrazolium blue (NTZB) reduction to a colored product, formazan. The maximal absorption peak of the colored product is near λ = 525 nm and the linear range is observed in the interval 0.005–0.14 mM of L-lactate. The main advantages of the proposed method when compared to the LDH-based routine approaches are a higher sensitivity (2.0 μM of L-lactate), simple procedure of analysis, usage of inexpensive, nontoxic reagents, and small amount of the enzyme. Enzymatic oxidation of L-lactate catalyzed by flavocytochrome b (2) and coupled with formazan production from nitrotetrazolium blue was shown to be used for L-lactate assay in food samples. A high correlation between results of the proposed method and reference ones proves the possibility to use flavocytochrome b (2)-catalysed reaction for enzymatic measurement of L-lactate in biotechnology and food chemistry. Hindawi Publishing Corporation 2013-09-24 /pmc/articles/PMC3800577/ /pubmed/24223505 http://dx.doi.org/10.1155/2013/461284 Text en Copyright © 2013 Oleh Smutok et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Smutok, Oleh Karkovska, Maria Smutok, Halyna Gonchar, Mykhailo Flavocytochrome b(2)-Based Enzymatic Method of L-Lactate Assay in Food Products |
title | Flavocytochrome b(2)-Based Enzymatic Method of L-Lactate Assay in Food Products |
title_full | Flavocytochrome b(2)-Based Enzymatic Method of L-Lactate Assay in Food Products |
title_fullStr | Flavocytochrome b(2)-Based Enzymatic Method of L-Lactate Assay in Food Products |
title_full_unstemmed | Flavocytochrome b(2)-Based Enzymatic Method of L-Lactate Assay in Food Products |
title_short | Flavocytochrome b(2)-Based Enzymatic Method of L-Lactate Assay in Food Products |
title_sort | flavocytochrome b(2)-based enzymatic method of l-lactate assay in food products |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3800577/ https://www.ncbi.nlm.nih.gov/pubmed/24223505 http://dx.doi.org/10.1155/2013/461284 |
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