Líffræðifélag Íslands
Líffræðiráðstefnan 2015
Erindi/veggspjald / Talk/poster E47
Fridjonsson OH (1), Hreggvidsson GO (1,2), Ævarsson A (3), Skírnisdottir S (1), Hjörleifsdottir S (4), Thorisdottir A (5), Pétursdóttir S (1), Ólafsdóttir S (1), Olgudóttir E (1), Ellertsson BÖ (1), Björnsdóttir S (1,2), Plotka M (6), Kaczorowska AK (6), Morzywolek A (6), Makowska J (6), Kozlowski LP (6), Dabrowski S (7), Bujnicki JM (6), Kaczorowski T (6), Kristjansson JK (2)
"1. Matís ohf; 2. Faculty of Life and Environmental Sciences, University of Iceland; 3. Prokazyme ltd; 4. Orf Genetics; 5. VESO Vikan, Namsos, Norway; 6. Department of Microbiology, University of Gdansk, Gdansk, Poland; 7. A&A Biotechnology, Gdynia, Poland."
Kynnir / Presenter: Ólafur H. Friðjónsson
Tengiliður / Corresponding author: Ólafur H. Friðjónsson (olafur@matis.is)
Many phage enzymes are key tools in molecular biology. As high temperatures may give a clear advantage in many DNA/RNA applications, thermophilic phages are of great interest as a source of thermostable molecular enzymes. The main objective of the EU project EXGENOMES was to develop new and improved thermostable enzymes for use in molecular biology and large-scale DNA synthesis. The target sources for the new enzymes were genome sequences of 18 thermophilic phages from Matís proprietary phage collection, isolated and sequenced in previous projects. Furthermore, prophages, plasmids, transposons and phage-like genes in 15 bacterial genomes from the genus Thermus were analyzed for this purpose. In addition about 2000 liters of hot spring water were filtered and fractionated by cytometry, resulting in two viral DNA metagenomes that were analyzed for new genes encoding molecular enzymes. 40 genes encoding novel enzymes were cloned and expressed and selected proteins were produced on a small scale and purified for activity evaluation and characterization. These included: helicase, RNAseH, lysozymes, RecA, DNA polymerases, primases, single strand binding proteins, and prototelomerase. Subsequently, 14 enzymes were obtained in pure and active form and taken for detailed characterization, protocol design and prototype construction and finally 6 enzymes were developed into commercial products with product sheets and QC. Some of these enzymes are currently commercially available.