Kemp elimination catalysts by computational enzyme design Export

@article{citeulike:2568656, abstract = {The design of new enzymes for reactions not catalysed by naturally occurring biocatalysts is a challenge for protein engineering and is a critical test of our understanding of enzyme catalysis. Here we describe the computational design of eight enzymes that use two different catalytic motifs to catalyse the Kemp elimination—a model reaction for proton transfer from carbon—with measured rate enhancements of up to 105 and multiple turnovers. Mutational analysis confirms that catalysis depends on the computationally designed active sites, and a high-resolution crystal structure suggests that the designs have close to atomic accuracy. Application of in vitro evolution to enhance the computational designs produced a >200-fold increase in kcat/Km (kcat/Km of 2,600 M-1s-1 and kcat/kuncat of >106). These results demonstrate the power of combining computational protein design with directed evolution for creating new enzymes, and we anticipate the creation of a wide range of useful new catalysts in the future.}, author = {Rothlisberger, Daniela and Khersonsky, Olga and Wollacott, Andrew M. and Jiang, Lin and DeChancie, Jason and Betker, Jamie and Gallaher, Jasmine L. and Althoff, Eric A. and Zanghellini, Alexandre and Dym, Orly and Albeck, Shira and Houk, Kendall N. and Tawfik, Dan S. and Baker, David}, citeulike-article-id = {2568656}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nature06879}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nature06879}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/18354394}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=18354394}, day = {19}, doi = {10.1038/nature06879}, issn = {0028-0836}, journal = {Nature}, keywords = {clip2, design, enzyme, protein}, month = {March}, number = {7192}, pages = {190--195}, posted-at = {2008-05-08 12:06:27}, priority = {2}, publisher = {Nature Publishing Group}, title = {Kemp elimination catalysts by computational enzyme design}, url = {http://dx.doi.org/10.1038/nature06879}, volume = {453}, year = {2008} }

TY - JOUR ID - citeulike:2568656 L3 - citeulike-article-id:2568656 N2 - The design of new enzymes for reactions not catalysed by naturally occurring biocatalysts is a challenge for protein engineering and is a critical test of our understanding of enzyme catalysis. Here we describe the computational design of eight enzymes that use two different catalytic motifs to catalyse the Kemp elimination—a model reaction for proton transfer from carbon—with measured rate enhancements of up to 105 and multiple turnovers. Mutational analysis confirms that catalysis depends on the computationally designed active sites, and a high-resolution crystal structure suggests that the designs have close to atomic accuracy. Application of in vitro evolution to enhance the computational designs produced a >200-fold increase in kcat/Km (kcat/Km of 2,600 M-1s-1 and kcat/kuncat of >106). These results demonstrate the power of combining computational protein design with directed evolution for creating new enzymes, and we anticipate the creation of a wide range of useful new catalysts in the future. IS - 7192 JF - Nature SN - 0028-0836 EP - 195 TI - Kemp elimination catalysts by computational enzyme design PB - Nature Publishing Group VL - 453 SP - 190 KW - clip2 KW - design KW - enzyme KW - protein AU - Rothlisberger, Daniela AU - Khersonsky, Olga AU - Wollacott, Andrew AU - Jiang, Lin AU - DeChancie, Jason AU - Betker, Jamie AU - Gallaher, Jasmine AU - Althoff, Eric AU - Zanghellini, Alexandre AU - Dym, Orly AU - Albeck, Shira AU - Houk, Kendall AU - Tawfik, Dan AU - Baker, David PY - 2008/03/19/ UR - http://dx.doi.org/10.1038/nature06879 ER -