Flavocytochrome b2-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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| Format: | Article |
| Language: | English |
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Wiley
2013-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1155/2013/461284 |
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| author | Oleh Smutok Maria Karkovska Halyna Smutok Mykhailo Gonchar |
| author_facet | Oleh Smutok Maria Karkovska Halyna Smutok Mykhailo Gonchar |
| author_sort | Oleh Smutok |
| collection | DOAJ |
| 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 b2 (EC 1.1.2.3; FC b2) 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 b2 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 b2-catalysed reaction for enzymatic measurement of L-lactate in biotechnology and food chemistry. |
| format | Article |
| id | doaj-art-df67b831bf4a4d08b918da535b0999db |
| institution | Kabale University |
| issn | 1537-744X |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-df67b831bf4a4d08b918da535b0999db2025-02-03T01:22:14ZengWileyThe Scientific World Journal1537-744X2013-01-01201310.1155/2013/461284461284Flavocytochrome b2-Based Enzymatic Method of L-Lactate Assay in Food ProductsOleh Smutok0Maria Karkovska1Halyna Smutok2Mykhailo Gonchar3Department of Analytical Biothecnology, Institute of Cell Biology, Drahomanov Street 14/16, Lviv 79005, UkraineDepartment of Analytical Biothecnology, Institute of Cell Biology, Drahomanov Street 14/16, Lviv 79005, UkraineDepartment of Analytical Biothecnology, Institute of Cell Biology, Drahomanov Street 14/16, Lviv 79005, UkraineDepartment of Analytical Biothecnology, Institute of Cell Biology, Drahomanov Street 14/16, Lviv 79005, UkraineL-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 b2 (EC 1.1.2.3; FC b2) 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 b2 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 b2-catalysed reaction for enzymatic measurement of L-lactate in biotechnology and food chemistry.http://dx.doi.org/10.1155/2013/461284 |
| spellingShingle | Oleh Smutok Maria Karkovska Halyna Smutok Mykhailo Gonchar Flavocytochrome b2-Based Enzymatic Method of L-Lactate Assay in Food Products The Scientific World Journal |
| title | Flavocytochrome b2-Based Enzymatic Method of L-Lactate Assay in Food Products |
| title_full | Flavocytochrome b2-Based Enzymatic Method of L-Lactate Assay in Food Products |
| title_fullStr | Flavocytochrome b2-Based Enzymatic Method of L-Lactate Assay in Food Products |
| title_full_unstemmed | Flavocytochrome b2-Based Enzymatic Method of L-Lactate Assay in Food Products |
| title_short | Flavocytochrome b2-Based Enzymatic Method of L-Lactate Assay in Food Products |
| title_sort | flavocytochrome b2 based enzymatic method of l lactate assay in food products |
| url | http://dx.doi.org/10.1155/2013/461284 |
| work_keys_str_mv | AT olehsmutok flavocytochromeb2basedenzymaticmethodofllactateassayinfoodproducts AT mariakarkovska flavocytochromeb2basedenzymaticmethodofllactateassayinfoodproducts AT halynasmutok flavocytochromeb2basedenzymaticmethodofllactateassayinfoodproducts AT mykhailogonchar flavocytochromeb2basedenzymaticmethodofllactateassayinfoodproducts |