Ingenza - Technology

Directed Evolution of Enzyme Activity and Specificity

In order to rapidly produce enzymes with useful activities it is important to have the right toolbox of techniques and equipment on hand. Ingenza has spent considerable effort developing a range of high throughput screening methodologies, from readily adaptable robotic-based chromatography-coupled mass spectrometry screens to colorimetric plate assays and plate based microbial selections, that can screen libraries of up to 10¹⁰ members. This range of screens on top of a foundation of molecular biology and process chemistry allows Ingenza to identify and evolve a wide range of different enzyme classes for formulation into biocatalysts and readily develop bioprocesses for new molecules and applications.

Of particular importance for the generation of oxidase biocatalysts for the broad application of the deracemisation technology is the ability to screen for variants with altered substrate specificity and improved activities. Using a colony-based colorimetric screen it is possible to screen around 10⁶ enzyme variants in each experiment. The screen relies on the detection of the hydrogen peroxide by-product of successful oxidase reactions and as such is applicable to the evolution of all related oxidase enzymes (including amine oxidases and amino acid oxidases).

Directed Evolution of Enzyme Activity and Specificity

The cycle of laboratory evolution of new oxidase specificities and improved performance.