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4a, b) such as Asp166Ala (abolishing interaction with the bound metal ion), Asp265Ala (removing H-bonding contacts with the galactose moiety), Ile266Gln (preventing rigid positioning of the galactose moiety in the GT1 cavity), did not produce any recombinant enzyme samples suitable for biochemical studies, and were consistent with previously published attempts describing a Asp166Ala-Asp168Ala double mutant, designed to abolish the GT1 DxD motif 34
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