Crystallographic characterization of the (R)-selective amine transaminase from Aspergillus fumigatus.
The importance of amine transaminases for producing optically pure chiral precursors for pharmaceuticals and chemicals has substantially increased in recent years. The X-ray crystal structure of the (R)-selective amine transaminase from the fungus Aspergillus fumigatus was solved by S-SAD phasing to 1.84 Å resolution. The refined structure at 1.27 Å resolution provides detailed knowledge about the molecular basis of substrate recognition and conversion to facilitate protein-engineering approaches. The protein forms a homodimer and belongs to fold class IV of the pyridoxal-5'-phosphate-dependent enzymes. Both subunits contribute residues to form two active sites. The structure of the holoenzyme shows the catalytically important cofactor pyridoxal-5'-phosphate bound as an internal aldimine with the catalytically responsible amino-acid residue Lys179, as well as in its free form. A long N-terminal helix is an important feature for the stability of this fungal (R)-selective amine transaminase, but is missing in branched-chain amino-acid aminotransferases and D-amino-acid aminotransferases.
Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489 Greifswald, Germany.
Maren
M
Thomsen
Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489 Greifswald, Germany.
Lilly
L
Skalden
Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489 Greifswald, Germany.
Gottfried J
GJ
Palm
Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489 Greifswald, Germany.
Matthias
M
Höhne
Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489 Greifswald, Germany.
Uwe T
UT
Bornscheuer
Institute of Biochemistry, University of Greifswald, Felix-Hausdorff-Strasse 4, 17489 Greifswald, Germany.
Winfried
W
Hinrichs
D001232:Aspergillus fumigatus / Q000201:enzymology*; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D011485:Protein Binding; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D040681:Structural Homology, Protein; D000637:Transaminases / Q000737:chemistry* / Q000378:metabolism
D000637:Transaminases
(R)-selective amine transaminase; S-SAD; pyridoxal phosphate