High-throughput screening for distinguishing nitrilases from nitrile hydratases in Aspergillus and application of a Box-Behnken design for the optimization of nitrilase.

Nitrilases and nitrile hydratases/amidases hydrolyze nitriles into carboxylic acids and/or amides, which are used in industrial chemical processes. In the present study, twenty-six microorganisms, including yeasts and filamentous fungi, in a minimum solid mineral medium supplemented with glucose and phenylacetonitrile were screened to evaluate their biocatalytic potential. Of these microorganisms, five fungi of the genus Aspergillus were selected and subjected to colorimetry studies to evaluate the production and distinction of nitrilase and nitrile hydratase/amidase enzymes. A. parasiticus Speare 7967 and A. niger Tiegh. 8285 produced nitrilases and nitrile hydratase, respectively. Nitrilase optimization was performed using a Box-Behnken design (BBD) and fungus A. parasiticus Speare 7967 with phenylacetonitrile volume (μL), pH, and carbohydrate source (starch:glucose; g/g) as independent variables and nitrilase activity (U mL-1 ) as dependent variable. Maximum activity (2.97×10-3 U mL-1 ) was obtained at pH 5.5, 80 μL of phenylacetonitrile, and 15 g of glucose. A. parasiticus Speare 7967 showed promise in the biotransformation of nitriles to carboxylic acids. Microbial screening of the species isolated from different places of Bahia-Brazil. Determination of the enzymatic pathway (nitrilase and/or nitrile hydratase/amidase) by colorimetric assay. Nitrilase of the Aspergillus parasiticus Speare 7967 optimized by Box-Behnken Design. This article is protected by copyright. All rights reserved.