By Cotor V., Alfonso I., Garcia-Urdiales E. (eds.)
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Extra resources for Asymmetric Organic Synthesis with Enzymes
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The formation of focused mutant libraries also provides the experimenter with a useful tool in directed evolution . To apply such methods, information regarding the structure of the enzyme (X-ray data or homology model), may be necessary. An intelligent decision is then made regarding the identiﬁcation of a hot spot, a position in the amino acid sequence of the enzyme that is believed to be important with respect to a given catalytic property, for example, near the binding pocket. Following this choice, the position is randomized by saturation mutagenesis leading to an enzyme library containing the 20 theoretically different mutants corresponding to the 20 proteinogenic amino acids at a deﬁned position [7,17].
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