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

Abstract

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 lM to 24lM with the detection limit of 0.25 lM. For ADI-OPA-S variant of the assay, the linearity varies from 0.7 lM to 50 lM with the detection limit of 0.55 lM. 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.