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Targeted Libraries and their Applications for Directed Evolution of Enzymes

Recorded event — October 28, 2020

Cellibre is a next-generation cellular agriculture company working on producing cannabinoids in an advantaged host. One of their first milestones was discovering and engineering a novel enzyme for the synthesis of CBGA, the molecule from which most cannabinoids are derived. Using gene synthesis, they quickly screened a panel of 88 novel enzymes identified by traditional and artificial intelligence-based discovery methods. Five of these showed potential for CBGA production, and one was selected for engineering. After modeling the enzyme’s structure, they identified active site residues and performed iterative rounds of saturation mutagenesis by leveraging the BioXp™ system — the world’s only synthetic biology workstation — for fast and efficient library construction. For the latest round of engineering, they combined beneficial mutations in specific combinations. This highly focused library was made possible by adapting the workflow for targeted libraries for which hundreds of specific constructs were delivered individually. Compared to the wild-type enzyme, their current top engineered variant has significantly improved specificity — from 11% to over 98% — and a 6-fold increase in total activity. They are currently designing even more diverse, complex libraries that would not be possible without targeted libraries.

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