|Title||Computer-assisted Recombination (CompassR) teaches us how to recombine beneficial substitutions from directed evolution campaigns.|
|Publication Type||Journal Article|
|Year of Publication||2019|
|Authors||Cui H, Cao H, Cai H, Karl-Erich J, Davari MDolatabadi, Schwaneberg U|
|Date Published||2019 Sep 25|
A main remaining challenge in protein engineering is how to recombine beneficial substitutions. Systematic recombination studies show that poorly performing variants are usually obtained after recombination of 3 to 4 beneficial substitutions. This limits researchers to exploit nature's potential in generating better enzymes. The Computer-assisted Recombination (CompassR) strategy provides a selection guide for beneficial substitutions that can be recombined to gradually improve enzyme performance by analysis of the relative free energy of folding (ΔΔG fold ). The performance of CompassR was evaluated by analysis of 84 recombinants located on 13 positions of Bacillus subtilis lipase A. The finally obtained variant F17S/V54K/D64N/D91E had a 2.7-fold improved specific activity in 18.3% (v/v) 1-Butyl-3-methylimidazolium Chloride ([BMIM][Cl]). In essence, the deducted CompassR rule allows to recombine beneficial substitutions in an iterative manner and empowers researchers to generate better enzymes in a time-efficient manner.