Recombination strategies 2GenReP and InSiReP employing CompassR yield highly organic solvent-tolerant enzymes


Congratulations Haiyang Cui on his recent publication!

In Haiyang(Ocean)´s new publication, 15 beneficial substitutions from Bacillus subtilis lipase A (BSLA), which improve organic cosolvent 1,4-dioxane resistance, were studied to compare two recombination strategies 2GenReP (two gene recombination process) and InSiReP (in silico guided recombination process) employing CompassR (Computer-assisted Recombination). CompassR was developed as a selection tool to identify beneficial substitutions that are compatible and can therefore likely be combined in a beneficial manner. However, the question of how to efficiently explore the protein sequence space when ten or more beneficial substitutions are identified has not yet been addressed. In this work, two strategies, 2GenReP and InSiReP, to recombine beneficial substitutions selected by CompassR analysis are compared with the goal of minimizing experimental efforts and maximizing property improvements. A remarkable multiple organic solvent resistant BSLA variant with 6 substitutions (up to 14.6-fold stability improvement in organic cosolvents) was obtained by both strategies after screening of less than 300 clones.

This work was realized in the division Computational Biology and was supported by computing resources granted by JARA-HPC from RWTH Aachen University (JARA0169). Haiyang Cui is financially supported by the China Scholarship Council (CSC) scholarship.

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Cui, H., Jaeger, K., Davari, M..D. and Schwaneberg, U. (2020), CompassR yields highly organic solvent‐tolerant enzymes through recombination of compatible substitutions. Chem. Eur. J.. Accepted Author Manuscript.

  Overview of 2GenReP and InSiReP recombination strategies. Copyright: © Bio VI  

Figure Overview of 2GenReP and InSiReP recombination strategies.