New DNA Polymerases
Sygnis is actively working on the discovery and characterization of novel DNA polymerases of interest for biotechnological applications. Moreover, Sygnis also develops improved versions of the enzymes currently included in Sygnis’ portfolio:
Sygnis has identified a novel DNA polymerase derived from an animal ancestor with similarity to Phi29 polymerase and possible improved properties for DNA amplification and sequencing. It could be a next-generation DNA polymerase and give SYGNIS a unique position on the market. The enzyme shows structural properties that support a great strand displacement capacity as well as an extraordinary processivity. Additionally, the enzyme maintains the active site responsible for the proofreading activity, guaranteeing high synthesis fidelity. These characteristics are essential to perform isothermal multiple displacement amplification and sequencing, making this enzyme a very attractive candidate to improve current sequencing and amplification technologies based on Phi29 DNA polymerase.
Sygnis concurrently works in finding also new PrimPol enzymes with interesting and improved properties. On one hand, Sygnis has already identified new PrimPol enzymes from viruses and microorganisms living in extreme environments (e.g. an Antarctic lake). Sequence analysis, together with their adaptation to an extreme environment, indicate that these enzymes could show interesting properties from a biotechnological point of view. For instance, they could have an enhanced affinity for substrates due to energy restrictions. On the other hand, the availability of primary sequences from PrimPol orthologs in Bacteria and Archaea, together with the extrapolation of the 3D-structure from related enzymes, has enabled Sygnis to identify TthPrimPol aminoacid residues involved in dNTP and DNA binding as well as in primase and polymerase catalysis. A collection of single-point mutations has been designed to improve dNTP and/or DNA affinity, and to expand TthPrimPol initiation site sequence to improve the generation of DNA primers in all sequence contexts. Furthermore, truncated versions of the enzyme have been also designed to enhance enzyme stability, preserving the enzymatic activity. Improved versions of TthPrimPol will be incorporated into upgraded TruePrime™ series of kits in the near future.