1NX9

Acetobacter turbidans alpha-amino acid ester hydrolase S205A mutant complexed with ampicillin

Summary for 1NX9

• Entry DOI: 10.2210/pdb1nx9/pdb
• Descriptor: alpha-amino acid ester hydrolase, (2S,5R,6R)-6-{[(2R)-2-AMINO-2-PHENYLETHANOYL]AMINO}-3,3-DIMETHYL-7-OXO-4-THIA-1-AZABICYCLO[3.2.0]HEPTANE-2-CARBOXYLIC ACID, GLYCEROL, ... (4 entities in total)
• Functional Keywords: alpha/beta hydrolase, jellyroll, hydrolase
• Biological source: Acetobacter pasteurianus
• Total number of polymer chains: 4
• Total formula weight: 293491.87

Authors

Barends, T.R.M.,Polderman-Tijmes, J.J.,Jekel, P.A.,Janssen, D.B.,Dijkstra, B.W. (deposition date: 2003-02-10, release date: 2004-03-09, Last modification date: 2024-02-14)

Primary citation

Barends, T.R.M.,Polderman-Tijmes, J.J.,Jekel, P.A.,Williams, C.,Wybenga, G.,Janssen, D.B.,Dijkstra, B.W.
Acetobacter turbidans {alpha}-Amino Acid Ester Hydrolase: HOW A SINGLE MUTATION IMPROVES AN ANTIBIOTIC-PRODUCING ENZYME.
J.Biol.Chem., 281:5804-5810, 2006

PubMed Abstract: The alpha-amino acid ester hydrolase (AEH) from Acetobacter turbidans is a bacterial enzyme catalyzing the hydrolysis and synthesis of beta-lactam antibiotics. The crystal structures of the native enzyme, both unliganded and in complex with the hydrolysis product D-phenylglycine are reported, as well as the structures of an inactive mutant (S205A) complexed with the substrate ampicillin, and an active site mutant (Y206A) with an increased tendency to catalyze antibiotic production rather than hydrolysis. The structure of the native enzyme shows an acyl binding pocket, in which D-phenylglycine binds, and an additional space that is large enough to accommodate the beta-lactam moiety of an antibiotic. In the S205A mutant, ampicillin binds in this pocket in a non-productive manner, making extensive contacts with the side chain of Tyr(112), which also participates in oxyanion hole formation. In the Y206A mutant, the Tyr(112) side chain has moved with its hydroxyl group toward the catalytic serine. Because this changes the properties of the beta-lactam binding site, this could explain the increased beta-lactam transferase activity of this mutant.

PubMed: 16377627
DOI: 10.1074/jbc.M511187200

Experimental method

X-RAY DIFFRACTION (2.2 Å)