Daniel Dourado / Almac group, Craigavon, United Kingdom
Alexandra Carvalho / Almac group, Craigavon, United Kingdom
Lyndsey Bannon / Almac Group, Craigavon, United Kingdom
Derek Quinn / Almac Group, Craigavon, United Kingdom
Stefan Mix / Almac Group, Craigavon, United Kingdom
Thomas Moody / Almac Group, Craigavon, United Kingdom
Topic: Industrial Biocatalysis
Enzyme engineering has been increasingly transforming the chemical and GMP API manufacture landscape. In-silico methods are responsible for the development of a new generation of improved biocatalysts by exploring the enzymes multidimensional fitness landscapes, obtaining variants at a fraction of the time and cost. Three examples will be presented. A methionine sulfoxide reductase was rational engineered to accept bulky sulfoxides. The best variant was found to catalyze these sulfoxides kinetic resolution with an ee up to 99%. A cyclase was first discovered and later engineered to produce Ambrox. For a reaction scale of 50g/L of substrate and 2.5% of enzyme loading, some variants show 142-fold improvement in activity over the WT. Finally, a nitrilase was engineered for one-pot, enantioselective, dynamic kinetic resolution for obtaining (R)-2-methoxymandelic acid. After process development an ee of 97% and 70% isolated yield were obtained on multigram scale.
