金属β-内酰胺酶CcrA的热力学研究.docVIP

  Thermokinetic characterization of imipenem hydrolysis with metallo-β-lactamase CcrA from Bacteroides fragilis# Zhai Le, Yang Kewu, Liu Chengcheng, Gao Huizhou, Yang Xia, Shi Ying, Wen Jing ** 5 10 15 20 25 (Northwest University, Xian 710069) Abstract: Imipenem is a widely used carbapenem antibiotic clinically. In an effort to probe the antibiotic resistance in bacteria, we first report in this paper the thermokinetic parameters of the imipenem hydrolysis with B1 subclass metallo-β-lactamase CcrA from Bacteroides fragilis. The values of activation free energy are 87.540 ± 0.027, 88.781 ± 0.031, 89.901 ± 0.036, and 91.057 ± 0.021 kJ·mol-1 at 293.15, 298.15, 303.15, and 308.15 K, respectively, the activation enthalpy is 19.099 ± 0.005 kJ·mol-1, the activation entropy is -233.57 ± 0.09 J·mol-1·K-1 and the apparent activation energy E is 21.598 kJ·mol-1. The results indicated that the imipenem hydrolysis by CcrA was an exothermic and spontaneous reaction with order of 1.4 and had higher activation enthalpy and lower activation entropy than the cefazolin hydrolysis. Keywords: Microcalorimetry;Metallo-β-lactamase CcrA;Antibiotic hydrolysis;Imipenem;Thermokinetic parameters 0 Introduction The β-lactam containing compounds, one of the most useful antibiotics which covalently modify peptidoglycan biosynthetic enzymes responsible for catalyzing the final transpeptidation step of cell wall biosynthesis, are the most preferred treatment regimens for many infectious diseases [1, 2]. In this antibiotic series, the cyclic amide has been substituted with various functional groups, thereby creating a family of active compounds with different specificities such as penicillins, cephalosporins, and carbapenems [3-5]. Due to the outstanding sterilization effect, the β-lactam containing antibiotics have been widely used clinically. However, the overuse of these antibiotics have resulted large number of antibiotic resistant bacteria. The most prevalent resistance mechanism is the exp