Reetz, Manfred T. and Prasad, Shreenath and Carballeira, Jose D. and Gumulya, Yosephine and Bocola, Marco (2010) Iterative Saturation Mutagenesis Accelerates Laboratory Evolution of Enzyme Stereoselectivity: Rigorous Comparison with Traditional Methods. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 132 (26). pp. 9144-9152. ISSN 0002-7863,
Full text not available from this repository.Abstract
Efficacy in laboratory evolution of enzymes is currently a pressing issue, making comparative studies of different methods and strategies mandatory Recent reports indicate that iterative saturation mutagenesis (ISM) provides a means to accelerate directed evolution of stereoselectivity and thermostability, but statistically meaningful comparisons with other methods have not been documented to date In the present study, the efficacy of ISM has been rigorously tested by applying it to the previously most systematically studied enzyme in directed evolution, the lipase from Pseudomonas aeruginosa as a catalyst in the stereoselective hydrolytic kinetic resolution of a chiral ester Upon screening only 10 000 transformants, unprecedented enantioselectivity was achieved (E = 594). ISM proves to be considerably more efficient than all previous systematic efforts utilizing error-prone polymerase chain reaction at different mutation rates, saturation mutagenesis at hot spots, and/or DNA shuffling, pronounced positive epistatic effects being the underlying reason
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PSEUDOMONAS-AERUGINOSA LIPASE; TOLUENE ORTHO-MONOOXYGENASE; CANDIDA-ANTARCTICA LIPASE; DIRECTED EVOLUTION; ACTIVE-SITE; BINDING-SITE; FORCE-FIELD; ENANTIOSELECTIVE ENZYMES; ORGANIC-CHEMISTRY; PROTEIN LIBRARIES; |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biophysik und physikalische Biochemie |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 22 Jul 2020 10:49 |
| Last Modified: | 22 Jul 2020 10:49 |
| URI: | https://pred.uni-regensburg.de/id/eprint/24466 |
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