By Brakmann S. (ed.), Johnson K. (ed.)
Fourteen chapters supply a image of a few of the hot advances within the software of evolutionary and combinatorial concepts within the improvement of proteins. After outlining the basic rules of the sector, papers handle such themes because the directed evolution of binding proteins by way of mobilephone floor reveal, screening recommendations of biocatalyst discovery, directed evolution as a way to create enantioselective enzymes to be used in natural chemistry, and utilized molecular evolution of enzymes fascinated about synthesis and service of DNA.
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Extra resources for Directed Molecular Evolution of Proteins
An engineered E. coli selection system lacking endogenous chorismate mutase activity . The genes encoding the bifunctional enzymes chorismate mutase-prephenate dehydrogenase and chorismate mutase-prephenate dehydratase were deleted, and monofunctional versions of the dehydrogenase and dehydratase were supplied on plasmid pKIMP-UAUC. Poten- tial chorismate mutases from random gene libraries are evaluated on the basis of their ability to complement the genetic defect and allow the cells to grow in the absence of added tyrosine (Tyr) and phenylalanine (Phe).
For instance, Arg90 might be replaced with lysine or methionine to examine what happens when the guanidinium is replaced with a different cation or when the positive charge is removed completely. But because a selection system is available, all 20 natural amino acids can be evaluated at position 90 simultaneously. Those amino acids that yield a functional enzyme can be quickly identified by sequencing the genes able to complement the chorismate mutase deficiency. When this experiment was performed, the results were dramatic .
F. Stadler, J. Theor. Biol, 2001, 212, 35 ± 40.  S. Govindarajan, R. A. Goldstein, Biopolymers, 1997, 42, 427 ± 438.  S. Govindarajan, R. A. Goldstein, Proteins, 1997, 29, 461 ± 466.  S. Gavrilets, Trends in Ecology and Evolution, 1997, 12, 307 ± 312.  S. Dalal, S. Balasubramanian, L. Regan, Nat. Struct. , 1997, 4, 548 ± 552. Directed Molecular Evolution of Proteins: or How to Improve Enzymes for Biocatalysis. Edited by Susanne Brakmann and Kai Johnsson Copyright ã 2002 Wiley-VCH Verlag GmbH & Co.
Directed Molecular Evolution of Proteins by Brakmann S. (ed.), Johnson K. (ed.)