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Plasmid DNA Preparation> | … the incorporation at the N-terminus of the mature Pif97 sequence a Trx-His tag and a TEV cleavage site (Figure S1), and this combined sequence was subcloned into the expression vector E4 plasmid for E coli expression as performed by GenScript USA (Piscataway, NJ) using … | Get A Quote |
(R)-stereospecific amine transaminases (R-ATAs) are important biocatalysts for the production of (R)-amine compounds in a strict stereospecific manner. An improved R-ATA, ATA-117-Rd11, was successfully engineered for the manufacture of sitagliptin, a widely used therapeutic agent for type-2 diabetes. The effects of the individual mutations, however, have not yet been demonstrated due to the lack of experimentally determined structural information. Here we describe three crystal structures of the first isolated R-ATA, its G136F mutant and engineered ATA-117-Rd11, which indicated that the mutation introduced into the 136(th) residue altered the conformation of a loop next to the active site, resul... More