  Our previously presented method for high throughput computational screening
of mutant activity (Hediger et al., arXiv:1203.2950) is benchmarked against
experimentally measured amidase activity for 22 mutants of Candida antarctica
lipase B (CalB). Using an appropriate cutoff criterion for the computed
barriers, the qualitative activity of 15 out of 22 mutants is correctly
predicted. The method identifies four of the six most active mutants with
>=3-fold wild type activity and seven out of the eight least active mutants
with <=0.5-fold wild type activity. The method is further used to screen all
sterically possible (386) double-, triple- and quadruple-mutants constructed
from the most active single mutants. Based on the benchmark test at least 20
new promising mutants are identified.
