3WWJ

Crystal structure of an engineered sitagliptin-producing transaminase, ATA-117-Rd11

Summary for 3WWJ

• Entry DOI: 10.2210/pdb3wwj/pdb
• Related: 3WWH 3WWI
• Descriptor: (R)-amine transaminase, PYRIDOXAL-5'-PHOSPHATE (3 entities in total)
• Functional Keywords: transaminase, amine-pyruvate aminotransferase, pyridoxal-5'-phosphate, multi-domain protein (alpha and beta), fold class iv plp-dependent enzyme, transferase
• Biological source: Arthrobacter sp. KNK168
• Total number of polymer chains: 12
• Total formula weight: 442250.32

Authors

Guan, L.J.,Ohtsuka, J.,Okai, M.,Miyakawa, T.,Mase, T.,Zhi, Y.,Hou, F.,Ito, N.,Yasohara, Y.,Tanokura, M. (deposition date: 2014-06-18, release date: 2015-08-12, Last modification date: 2018-11-21)

Primary citation

Guan, L.J.,Ohtsuka, J.,Okai, M.,Miyakawa, T.,Mase, T.,Zhi, Y.,Hou, F.,Ito, N.,Iwasaki, A.,Yasohara, Y.,Tanokura, M.
A new target region for changing the substrate specificity of amine transaminases.
Sci Rep, 5:10753-10753, 2015

PubMed Abstract: (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, resulting in a substrate-binding site with drastically modified volume, shape, and surface properties, to accommodate the large pro-sitagliptin ketone. Our findings provide a detailed explanation of the previously reported molecular engineering of ATA-117-Rd11 and propose that the loop near the active site is a new target for the rational design to change the substrate specificity of ATAs.

PubMed: 26030619
DOI: 10.1038/srep10753

Experimental method

X-RAY DIFFRACTION (2.2 Å)