As part of our ongoing nitrilase discovery project, we are trying a panel of about 50 substrates of various types against an ever-growing panel of nitrilases. The first round panel has 20 enzymes, and using a chromogenic assay we are seeing the rather nice selectivity that the literature indicates these enzymes show. Below, an example of two nitrilases (with each repeated) and their response to 4 simple nitriles- the depth of colour scales nicely with the amount of ammonia released by the nitrile hydrolysis.
There are similar levels of diversity of response for aryl over alkyl nitriles, and short chain over long chain nitriles. After the colour screening we are moving onto GCMS for confirmatory results, as well as doing a bit of NMR for that final structural evidence.
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