3N5F

Crystal Structure of L-N-carbamoylase from Geobacillus stearothermophilus CECT43

Summary for 3N5F

• Entry DOI: 10.2210/pdb3n5f/pdb
• Descriptor: N-carbamoyl-L-amino acid hydrolase, ISOPROPYL ALCOHOL, COBALT (II) ION, ... (5 entities in total)
• Functional Keywords: carbamoylase, hinge domain, m20 peptidase family, evolution, binding residue, dimerization domain, hydrolase
• Biological source: Bacillus stearothermophilus (Geobacillus stearothermophilus)
• Total number of polymer chains: 2
• Total formula weight: 88715.67

Authors

Garcia-Pino, A.,Martinez-Rodriguez, S.,Gavira, J.A. (deposition date: 2010-05-25, release date: 2011-05-25, Last modification date: 2023-09-06)

Primary citation

Martinez-Rodriguez, S.,Garcia-Pino, A.,Las Heras-Vazquez, F.J.,Clemente-Jimenez, J.M.,Rodriguez-Vico, F.,Garcia-Ruiz, J.M.,Loris, R.,Gavira, J.A.
Mutational and structural analysis of L-N-carbamoylase reveals new insights into a peptidase m20/m25/m40 family member.
J.Bacteriol., 194:5759-5768, 2012

PubMed Abstract: N-Carbamoyl-L-amino acid amidohydrolases (L-carbamoylases) are important industrial enzymes used in kinetic resolution of racemic mixtures of N-carbamoyl-amino acids due to their strict enantiospecificity. In this work, we report the first L-carbamoylase structure belonging to Geobacillus stearothermophilus CECT43 (BsLcar), at a resolution of 2.7 Å. Structural analysis of BsLcar and several members of the peptidase M20/M25/M40 family confirmed the expected conserved residues at the active site in this family, and site-directed mutagenesis revealed their relevance to substrate binding. We also found an unexpectedly conserved arginine residue (Arg(234) in BsLcar), proven to be critical for dimerization of the enzyme. The mutation of this sole residue resulted in a total loss of activity and prevented the formation of the dimer in BsLcar. Comparative studies revealed that the dimerization domain of the peptidase M20/M25/M40 family is a "small-molecule binding domain," allowing further evolutionary considerations for this enzyme family.

PubMed: 22904279
DOI: 10.1128/JB.01056-12

Experimental method

X-RAY DIFFRACTION (2.75 Å)