Optimalisering: MarSynth - Marine DNA modifying enzymes for synthetic biology

Synthetic biology is a rapidly evolving field, and is heralded as a possible solution for the challenges in future bioeconomy and bioenergy. The ultimate vision of synthetic biology is to create new biological operating systems of cells that predictably are able to carry out useful tasks. One of the key steps in a synthetic biology pipeline is the assembly of DNA fragments into larger functional constructs often involving multiple assemblies. A current bottleneck is however the lack of a robust room-temperature method to do multiple DNA assemblies without time-consuming manual treatment steps. The background for this project is the large marine bioprospecting efforts in the Arctic, coupled with knowledge based and innovative efforts in a previous Biotek2021 project, leading to identification and characterization of several marine heat-labile DNA-modifying enzymes with a high potential for industrial and biotechnological purposes. The proposed project aims at developing enzymes specialized for performing multiple DNA fragment assembly at room temperature. Through Proof of Principle studies, protocol and formulations of the engineered enzymes will be optimized to ensure a quick, efficient and reliable DNA assembly platform. The best enzyme composition will be selected for Proof of Concept study. An important part of this project is to continuously engage with end users to ensure that our assembly platform meets customers need. Key elements of responsible research and innovation will be integrated throughout the project.

We have developed a new technology platform based on two unique enzymes originating from marine bioprospecting in Tromsø. This technology outperforms the existing solutions and satisfies a need in the fast growing synthetic biology market. The project has contributed to enhance the existing competence and relationship between various players in the marine biotechnological research and innovation community and industry in Tromsø. We believe that the results of this project will further put Norway, and Tromsø in particular, on the map for exploitation of cold-adapted marine resources.

The ultimate vision of synthetic biology is to create new biological operating systems of cells that predictably are able to carry out useful tasks. One of the key steps in a synthetic biology pipeline is the assembly of DNA fragments into larger functional constructs often involving multiple assemblies. A current bottleneck is however the lack of a robust room-temperature method to do sequential (or multiple) DNA assemblies without time consuming manual treatment steps. A new DNA assembly method able to bypass the current hurdles is therefore highly desired. The proposed project aims at developing a sequence independent isothermal room-temperature DNA assembly platform, with potential for contributing to automation of a complete Synthetic Biology Pipeline. An easy-to-use DNA assembly kit format will be developed to serve the R&D market. The background for this project is the large marine bioprospecting efforts in the Arctic, coupled with knowledge based and innovative efforts during an ongoing Biotek2021 project, leading to identification and characterization of several marine heat-labile DNA-modifying enzymes with a high potential for industrial and biotechnological purposes. Through comparative structural analysis and rational design, enzymes specialized for performing isothermal DNA assembly at room temperature will be developed. The enzymes´s intrinsic heat-lability are key features for the versatility of the method. Through Proof of Principle (PoP) studies, protocol and formulations of the engineered enzymes will be optimized to ensure a quick, efficient and reliable DNA assembly platform. The best enzyme composition will be selected for Proof of Concept (PoC) and a User Case study, where the complete lactose operon will be assembled using the developed technology thus enabling the synthetic engineered bacteria to grow and utilize lactose as a carbon source. Key elements of responsible research and innovation will be integrated throughout the project.