E!12757 COLDLIGASE

COLDLIGASE, a collaboration project between UiT -The Arctic University of Norway, the Danish company Legomics and the Norwegian Company ArcticZymes Technologies ASA, aims to develop DNA ligases enabling a gene synthesis technology platform. We have identified and successfully characterized a handful of enzymes that show ligase activity at cold temperature. We have solved X-ray structures of two psychrophilic ligases and used a structured guided rational design to mutate key residues to improve the performance of these enzymes. Crystal structures of these ligase variants show additional interaction with the DNA substrate and are currently under investigation to evaluate their ligation reaction performance.

Characterization of interesting ligases identified in this project and engineering thereof have resulted in a number of ligases and mutens with promising new activity and substrate selectivity. The two X-ray structures and the additional X-ray structures of the ligases resulted in an array of new mutants based on a structure guided design. These will be of potential use in other molecular applications and will expand the ArcticZymes’ portfolio of DNA modifying enzymes. Biochemical and structural characterization of the cold ligases have shown new findings and insights into the structure-function relationship of these enzymes. This has open up the possibility for UiT and ArcticZymes of filing a patent and publish the results in peer-reviewed journals.

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