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https://doi.org/10.37358/Rev.Chim.1949

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Revista de Chimie (Rev. Chim.), Year 2020, Volume 71, Issue 9, 285-294

https://doi.org/10.37358/RC.20.9.8339

Cosmin Stefan Mocanu, Ana-Simona Bocec, Vasile Robert Gradinaru, Dana-Teodora Anton-Paduraru

A Biochemical Method for Tyrosine Determination in Phenylketonuria Using a Colorimetric Enzymatic Approach

Abstract:

Phenylketonuria is a serious genetic disease caused by a deficiency of phenylalanine metabolism, an essential amino acid found in daily nutrition. This disorder is caused by the lack of a specific enzyme called phenylalanine hydroxylase which mediates the conversion of phenylalanine to tyrosine). Thus, after ingestion of phenyl alanine-rich proteins, the amino acid concentration increases considerably in the blood due to proteolysis. Genetic deffects of enzymes responsible for phenylalanine metabolic conversion were intensively studied. Among them the defect of gene encoding phenylalanine hydroxylase has a higher notoriety. This genetic defect is translated in an inactive enzyme constructs that impair the aminoacid hydroxylation. This physiological stage is also called hyperphenylalaninemia where slightly high levels of phenylalanine are noticed in the blood or urine. Consequently, the amino acid is converted to phenylpyruvic acid by transamination, the later displaying a particularly toxic effect against brain tissues. The aim of this study was to quantify tyrosine in the blood of patients suffering of phenylketonuria by an alternative enzymatic method. The tyrosinase used in this assay was extracted from commercial mushrooms (Agaricus bisporus) following the Haghbeen protocol with some modifications. Two chromatographic steps (molecular exclusion chromatography and ionic exchange chromatography) were used during the enzyme purification process. High purity samples were concentrated using ultrafiltration. The tyrosinase was screened by a classical enzymatic microplate assay having DOPA as a substrate. Finally the pure enzyme was used in order to quantify tyrosine from different standard solutions. The level of tyrosine from deproteinized serum samples was determined using a similar enzymatic strategy.
Keywords:
phenylketonuria; tyrosinase; enzyme extraction; chromatography; spectroscopy

Issue: 2020, Volume 71, Issue 9
Pages: 285-294
Publication date: 2020/9/5
https://doi.org/10.37358/RC.20.9.8339
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Creative Commons License
This article is published under the Creative Commons Attribution 4.0 International License
Citation Styles
Cite this article as:
MOCANU, C.S., BOCEC, A., GRADINARU, V.R., ANTON-PADURARU, D., A Biochemical Method for Tyrosine Determination in Phenylketonuria Using a Colorimetric Enzymatic Approach, Rev. Chim., 71(9), 2020, 285-294.

Vancouver
Mocanu CS, Bocec A, Gradinaru VR, Anton-Paduraru D. A Biochemical Method for Tyrosine Determination in Phenylketonuria Using a Colorimetric Enzymatic Approach. Rev. Chim.[internet]. 2020 Sep;71(9):285-294. Available from: https://doi.org/10.37358/RC.20.9.8339


APA 6th edition
Mocanu, C.S., Bocec, A., Gradinaru, V.R. & Anton-Paduraru, D. (2020). A Biochemical Method for Tyrosine Determination in Phenylketonuria Using a Colorimetric Enzymatic Approach. Revista de Chimie, 71(9), 285-294. https://doi.org/10.37358/RC.20.9.8339


Harvard
Mocanu, C.S., Bocec, A., Gradinaru, V.R., Anton-Paduraru, D. (2020). 'A Biochemical Method for Tyrosine Determination in Phenylketonuria Using a Colorimetric Enzymatic Approach', Revista de Chimie, 71(9), pp. 285-294. https://doi.org/10.37358/RC.20.9.8339


IEEE
C.S. Mocanu, A. Bocec, V.R. Gradinaru, D. Anton-Paduraru, "A Biochemical Method for Tyrosine Determination in Phenylketonuria Using a Colorimetric Enzymatic Approach". Revista de Chimie, vol. 71, no. 9, pp. 285-294, 2020. [online]. https://doi.org/10.37358/RC.20.9.8339


Text
Cosmin Stefan Mocanu, Ana-Simona Bocec, Vasile Robert Gradinaru, Dana-Teodora Anton-Paduraru,
A Biochemical Method for Tyrosine Determination in Phenylketonuria Using a Colorimetric Enzymatic Approach,
Revista de Chimie,
Volume 71, Issue 9,
2020,
Pages 285-294,
ISSN 2668-8212,
https://doi.org/10.37358/RC.20.9.8339.
(https://revistadechimie.ro/Articles.asp?ID=8339)
Keywords: phenylketonuria; tyrosinase; enzyme extraction; chromatography; spectroscopy


RIS
TY - JOUR
T1 - A Biochemical Method for Tyrosine Determination in Phenylketonuria Using a Colorimetric Enzymatic Approach
A1 - Mocanu, Cosmin Stefan
A2 - Bocec, Ana-Simona
A3 - Gradinaru, Vasile Robert
A4 - Anton-Paduraru, Dana-Teodora
JF - Revista de Chimie
JO - Rev. Chim.
PB - Revista de Chimie SRL
SN - 2668-8212
Y1 - 2020
VL - 71
IS - 9
SP - 285
EP - 294
UR - https://doi.org/10.37358/RC.20.9.8339
KW - phenylketonuria
KW - tyrosinase
KW - enzyme extraction
KW - chromatography
KW - spectroscopy
ER -


BibTex
@article{RevCh2020P285,
author = {Mocanu Cosmin Stefan and Bocec Ana-Simona and Gradinaru Vasile Robert and Anton-Paduraru Dana-Teodora},
title = {A Biochemical Method for Tyrosine Determination in Phenylketonuria Using a Colorimetric Enzymatic Approach},
journal = {Revista de Chimie},
volume = {71},
number = {9},
pages = {285-294},
year = {2020},
issn = {2668-8212},
doi = {https://doi.org/10.37358/RC.20.9.8339},
url = {https://revistadechimie.ro/Articles.asp?ID=8339}
}
 
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