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As seen for the Tyr265Ala variant, introduction of lysine at position 265 decreases the racemase activity of the enzyme by more than three orders of magnitude. Neither D- nor L-alanine was found to be a substrate for the Tyr265Lys mutant.
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As seen for the Tyr265Ala variant, introduction of lysine at position 265 decreases the racemase activity of the enzyme by more than three orders of magnitude. Neither D- nor L-alanine was found to be a substrate for the Tyr265Lys mutant.
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(2R,3S)-β-Phenylserine was purchased as a racemic mixture of D- and L-threo-β-phenylserine isomers. In the presence of the aldolases, only the D-isomer is converted to product (reference [3]). The erythro isomer, (2R,3R)- β-phenylserine, was synthesized from cinnamic acid in five steps following an asymmetric dihydroxylation strategy: H. C. Kolb, M. S. Van-Nieuwenhze, K. B. Sharpless, Chem. Rev. 1994, 94, 2483;
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34250770286
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The models of the enzyme complexes are based on earlier hybrid docking and ab initio calculations performed with aldimines of all four α-methyl-β-phenylserine diastereomers at the Tyr265Ala active site (reference [4, The (2R,3S, and (2R,3R)-β- phenylserine derivatives were superimposed on their α-methyl counterparts, and the additional point mutations (Tyr265Lys, Met134Phe, and Ile352Trp) were introduced with the MacPymol software Delano Scientific LLC, 2006, manually choosing the rotamers that exhibit no steric clashes
-
The models of the enzyme complexes are based on earlier hybrid docking and ab initio calculations performed with aldimines of all four α-methyl-β-phenylserine diastereomers at the Tyr265Ala active site (reference [4]). The (2R,3S)- and (2R,3R)-β- phenylserine derivatives were superimposed on their α-methyl counterparts, and the additional point mutations (Tyr265Lys, Met134Phe, and Ile352Trp) were introduced with the MacPymol software (Delano Scientific LLC, 2006), manually choosing the rotamers that exhibit no steric clashes.
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