Department of Chemistry, University of Copenhagen , Copenhagen , Denmark.
PeerJ. 2013 Aug 29;1:e145. doi: 10.7717/peerj.145. eCollection 2013.
Our previously presented method for high throughput computational screening of mutant activity (Hediger et al., 2012) 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.
我们之前提出的高通量计算筛选突变体活性的方法（Hediger 等人，2012 年）已经针对 22 种南极假丝酵母脂肪酶 B（CalB）突变体的实验测量酰胺酶活性进行了基准测试。使用计算出的势垒的适当截止标准，正确预测了 22 个突变体中的 15 个的定性活性。该方法确定了六个最活跃的突变体中的四个，其活性≥野生型的 3 倍，八个最不活跃的突变体中的七个，其活性≤野生型的 0.5 倍。该方法还用于筛选由最活跃的单突变体构建的所有空间可能的（386）双、三、四重突变体。根据基准测试，至少确定了 20 个有前途的新突变体。
