Novel arginine deiminase-based method to assay L-arginine in beverages

被引:24
|
作者
Stasyuk, N. Ye. [1 ]
Gayda, G. Z. [1 ]
Fayura, L. R. [1 ]
Boretskyy, Y. R. [1 ,2 ]
Gonchar, M. V. [1 ,3 ]
Sibirny, A. A. [1 ,4 ]
机构
[1] Natl Acad Sci Ukraine, Inst Cell Biol, Drahomanov Str 14-16, UA-79005 Lvov, Ukraine
[2] Lviv State Univ Phys Culture, Dept Biochem & Hyg, Kosciuszko St 11, UA-79000 Lvov, Ukraine
[3] Rzeszow Univ, Inst Appl Biotechnol & Basic Sci, Sokolowska Str 26, PL-36100 Kolbuszowa, Poland
[4] Rzeszow Univ, Dept Biotechnol & Microbiol, Cwiklinskiej 2, PL-35601 Rzeszow, Poland
关键词
L-Arginine; Arginine deiminase; Enzymatic assay; Beverages; Ethyl carbamate; HIGH-LEVEL EXPRESSION; LACTIC-ACID BACTERIA; ETHYL CARBAMATE; COLORIMETRIC DETERMINATION; NITROGEN-FERTILIZATION;
D O I
10.1016/j.foodchem.2016.01.093
中图分类号
O69 [应用化学];
学科分类号
081704 ;
摘要
A highly selective and sensitive enzymatic method for the quantitative determination of L-arginine (Arg) has been developed. The method is based on the use of recombinant bacterial arginine deiminase (ADI) isolated from the cells of a recombinant strain Escherichia coli and o-phthalaldehyde (OPA) as a chemical reagent. Ammonia, the product of the enzymatic digestion of Arg by ADI, reacts with OPA and forms in the presence of sulfite a product, which can be detected by spectrophotometry (S) and fluorometry (F). The linear concentration range for Arg assay in the final reaction mixture varies for ADI-OPA-F variant of the method from 0.35 mu M to 24 mu M with the detection limit of 0.25 mu M. For ADI-OPA-S variant of the assay, the linearity varies from 0.7 mu M to 50 mu M with the detection limit of 0.55 mu M. The new method was tested on real samples of wines and juices. A high correlation (R = 0.978) was shown for the results obtained with the proposed and the reference enzymatic method. (C) 2016 Elsevier Ltd. All rights reserved.
引用
收藏
页码:320 / 326
页数:7
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