A comparison of simultaneous OmniChange with iterative CAST for cpADH5 engineering
Congratulations to Yunus Ensari on his recent publication!
In Yunus´s new publication, the first comprehensive comparison regarding an improvement in enzymatic activity and required screening efforts during iterative and simultaneous site saturation mutagenesis experiment was performed. Screening of 7200 clones from 33 site saturation mutagenesis libraries (exploring 17 recombination paths) yielded the cpADH5 W286A variant with a 82-fold improved initial activity toward methyl 3- hydroxyhexanoate. Screening of 3500 clones from a single OmniChange library with four positions (C57, W116, L119, and W286; 1.8 % of the generated sequence space) yielded the cpADH5 C57V/W286S variant with a 108-fold improvement in initial activity toward methyl 3-hydroxyhexanoate. In conclusion, combination of high throughput screening system with simultaneous site saturation of four and more positions seems to be highly promising to take full advantage of nature’s diversity.
To learn more, please access the full paper on Publications & Patents and
Ensari, Y., Dhoke, G., Davari, M., Ruff, A. J. and Schwaneberg, U. (2018), A comparative reengineering study of cpADH5 through iterative and simultaneous multi‐site saturation mutagenesis. ChemBioChem. Accepted Author Manuscript. doi:10.1002/cbic.201800159