DIRECTED EVOLUTION: BRINGING THE CYCLE OF LIFE TO THE LABORATORY BENCH
Directed evolution is an engineering strategy used to customize enzymes for different purposes, ranging from industrial and environmental applications to drug discovery. Mimicking the processes of natural evolution in the laboratory, this method was pioneered by Prof. Frances H. Arnold, for which she was awarded the Nobel Prize in Chemistry 2018. Indeed, it is considered the most revolutionary protein engineering method known to date.
In this procedure, genetic diversity is first created by random mutagenesis and/or the recombination of parental genes. Once the mutant libraries are expressed in a suitable host organism, they can be screened in high-throughput platforms to search for specific biochemical traits, for example: activity under extreme pH or temperatures, changes in substrate preferences, heterologous expression, tolerance in the presence of strong inhibitors, etc. In this way, selective pressure can be strictly controlled by the researcher until the desired properties are attained.
We can help you to develop your directed evolution platforms as well as to design enzymes for a variety of applications, from bioremediation to novel green processes.
EVOSHUFFLER is the EvoEnzyme technological platform. The company’s hallmark has been designed around the DNA recombination machinery of Saccharomyces cerevisiae, which is fully exploited for mutant library creation and screening. Our broad repertoire of in vivo DNA recombination methods is ranging from classical DNA shuffling to focused evolution (e.g. MORPHING, SDR). In addition, cutting-edge computational methods based on rossetta design and phylogenetic inference are being applied to create smart mutant libraries and resurrect ancestral enzymes with promiscuous activities.
Some recent case studies of our products and genetic/computational tools can be seen at:
Gomez de Santos, P., Lazaro, S., Viña-Gonzalez, J., Hoang, M.D., Glieder, A., Hollmann, F. and Alcalde, M. (2020). Evolved peroxygenase-aryl alcohol oxidase fusions for self-sufficient oxyfunctionalization reactions. ACS Catalysis 10: 13524-13534. https://dx.doi.org/10.1021/acscatal.0c03029
Gomez-Fernandez, B., Risso, V.A., Rueda, A., Sanchez-Ruiz, J.M. and Alcalde, M. (2020). Ancestral resurrection and directed evolution of fungal Mesozoic laccases. Applied and Environmental Microbiology. 86:e00778-20. JOURNAL COVER & SPOTLIGHT. https://doi.org/10.1128/AEM.01732-20
Viña-Gonzalez, J. And Alcalde, M. (2020). In vivo site-directed recombination (SDR): An efficient tool to reveal beneficial epistasis. In: Methods in Enzymology 643. Tawfik, D. Ed. Academic Press Elsevier Inc, Cambridge, MA, EEUU. Pages 1-12. ISBN 978-0-12-821149-6. https://doi.org/10.1016/bs.mie.2020.04.021.