How can we turn naturally occurring enzymes into powerful tools for making medicines without spending months or huge budgets on protein engineering?
A new open-access study in iScience introduces RISE (Rapid In Vitro Semi-rational Engineering): a streamlined workflow that allows chemistry labs to quickly improve enzymes using simple, cell-free methods.
What is new is that RISE combines computer-guided design of promising enzyme variants with fast PCR-based DNA mutagenesis (no cloning needed) and cell-free protein synthesis (no living cells required). These steps are linked in short, iterative cycles of testing and improvement. Together, this approach makes enzyme engineering much faster, cheaper, and more accessible, even for non-specialist laboratories.
Using RISE, the team engineered a ketimine reductase enzyme and achieved:
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Inverted stereoselectivity in a key reaction (from >99% R to up to 80% S)
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Up to 400-fold higher activity in another reaction
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Successful gram-scale synthesis of important intermediates for ACE2 inhibitor drugs
All this was accomplished in just a few iterative rounds, starting from a poorly performing wild-type enzyme.
Biocatalysis has huge potential for greener chemical manufacturing but enzyme optimisation is often too slow and expensive in early drug development. However, RISE enables rapid creation of “good enough” biocatalysts that can demonstrate synthetic feasibility, helping enzymatic routes move forward instead of being abandoned.
In short, RISE lowers technical and financial barriers to enzyme engineering and opens the door for wider adoption of biocatalysis in chemical and pharmaceutical research.
The article Accessible biocatalyst development by rapid in vitro semi-rational engineering (RISE) of enzymes was published in iScience, Volume 29, Issue 1. Read the full article here.