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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...

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Autores principales: Smutok, Oleh, Karkovska, Maria, Smutok, Halyna, Gonchar, Mykhailo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2013
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.
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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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