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

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.