A simple, efficient and rapid screening technique for differentiating nitrile hydratase and nitrilase producing...

Area of Science:

• Biochemistry and Microbiology
• Enzyme Technology

Background:

• Nitrile hydrolyzing enzymes are crucial for converting nitriles to amides and acids.
• Identifying bacteria with these enzymes is important for industrial applications like acrylamide production.

Purpose of the Study:

• To develop and validate a rapid, dye-based screening method for nitrile-degrading bacteria.
• To differentiate between nitrile hydratase (NHase) and nitrilase producing microorganisms.

Main Methods:

• Screening of 108 bacterial strains from soil samples using a 96-well plate assay.
• Utilized bromothymol blue as an indicator for detecting amides and acids via color change.
• Employed nesslerization and quantitative assays for enzyme activity confirmation.

Main Results:

• Six bacterial strains exhibited NHase activity, and eleven showed nitrilase activity.
• The screening method successfully differentiated between NHase and nitrilase producers.
• Rhodococcus rhodochrous demonstrated the highest NHase activity in quantitative assays.

Conclusions:

• The developed dye-based method enables rapid and efficient screening of nitrile-degrading bacteria.
• This method is valuable for identifying potent microbial candidates for acrylamide and acrylic acid production from acrylonitrile.
• The study highlights the potential of soil bacteria in biocatalysis for industrial chemical synthesis.