P4 Hot investigations into Cold-Sourced Enzymes

P4 Hot investigations into Cold-Sourced Enzymes

Adele Williamson, Bjørn Altermark, Marcin Pierechod, Gro Bjerga, Hege Pedersen, Nils-Peder Willassen, Arne Smalås

P4 Hot investigations into Cold-Sourced Enzymes

In the MARZymes project we aim to identify novel psychrophilic enzymes from marine Arctic sources for use in industrial and biotechnological applications. Here we present data on a
variety of proteins at different stages of investigation within the project. The target proteins are identified by either functional or in silico screening of marine bacteria and metagenomic DNA collected at temperatures between 0 and 4 °C. Activities of interest include proteins with putative functions in DNA or protein modification and carbohydrate degradation. Cold adaptation imparts technologically exploitable properties to psychrophilic enzymes such as high rates of catalytic activity, low temperature optima and low temperatures of thermal inactivation; however high flexibility and low stability present challenges for their expression and purification. Protein overproduction for enzyme characterization is carried out in E. coli
where these challenges are overcome using a combination of cold-adapted inducer systems as well as traditional culture condition modifications, co-expression partners and fusion tags.
The temperature dependence of an enzyme’s activity is essential in evaluating its potential for a particular application. Biophysical characterization of the recombinant protein to determine thermal and chemical stability can give insight into which assay and storage conditions are most appropriate to test.

Poster not available