Enzymatic analysis of the effect of naturally occurring Leu138Pro mutation identified in SHV β-lactamase on hydrolysis of penicillin and ampicillin
1 Department of Infectious Diseases, College of Veterinary Medicine, KRF Zoonotic Disease Priority Research Institute, Brain Korea 21 for Veterinary Science, Seoul National University, Seoul 151-742, S. Korea
2 School of Chemical and Biological Engineering, Seoul National University, Seoul 151-744, S. Korea
BMC Microbiology 2011, 11:29 doi:10.1186/1471-2180-11-29Published: 4 February 2011
The aim of this study was to analyze the significance of leucine to proline substitution at position 138(Leu138Pro) on the hydrolysis of penicillin and ampicillin that we identified in the blaSHV gene of clinical Escherichia coli swine isolate.
Kinetic analysis of the mutant proteins showed that Km value of the purified L138P mutant was comparatively higher than SHV-1, SHV-33 and SHV-33(L138P) enzyme for penicillin and ampicillin. Docking simulation of the SHV-1 and SHV-(L138P) enzymes also confirmed that β-lactamases preferred penicillin to ampicillin and the SHV-1 had a higher binding affinity for antibiotics compared to the SHV-(L138P) and other mutants.
Our result demonstrated that L138P has a reduced role in penicillin and ampicillin hydrolyzing properties of SHV β-lactamases. These naturally occurring mutations rendering reduced function of the existing protein could trigger the emergence or acquisition of more effective alternative mechanisms for β-lactam hydrolysis.