The use of clade‐specific PCR assays to identify novel nitrilase genes from environmental isolates

Abstract

Abstract Nitrilase enzymes (EC 3.5.5.1) are responsible for the direct hydration of nitriles to their corresponding carboxylic acids and ammonia. The utilization of nitrilase enzymes in biocatalysis toward bio‐pharmaceuticals and industrial applications facilitates the move towards green chemistry. The body of research presented describes a novel clade‐specific touchdown PCR protocol for the detection of novel nitrilase genes. The presented study identified partial sequences of 15 novel nitrilase genes across 7 genera, with partial DNA sequence homology (%) displayed across an additional 16 genera. This research will prove valuable in the screening of microorganisms for the identification of novel clade‐specific nitrilase genes, with predicted enantioselective profiles as determined by their clade characterizations.

Keywords

• clade specific
• degenerate primers
• environmental isolates
• gene detection
• nitrilase
• touchdown PCR