TEM 116-type ESBL natural enzymes and kinetic characteristics of recombinant enzyme.docVIP

 PAGE \* MERGEFORMAT 23 TEM 116-type ESBL natural enzymes and kinetic characteristics of recombinant enzyme Author: Hsien-Ming Chen Xiushu LU Jian-xin Wang Yong-Liang Lou, SUN Chang-Gui Lu Yong-sui 【Abstract】 Objective To study the natural TEM  116-type ESBL enzymes and recombinant enzyme kinetic properties, and compare their differences. Methods Enzymatic detection of ultraviolet spectrophotometry antibiotic hydrolysis reaction to Lee  Wilson double-reciprocal equation improved data processing method for data processing, determination of the natural enzyme and recombinant enzyme Km, Vmax, and kcat. Observations of temperature and pH on the enzymatic reaction. Results are Lee  Wilson double-reciprocal equation improved data processing method for data processing convenient and accurate determination of the natural enzymes and recombinant enzyme Km and Vmax and calculate the kcat. Temperature and pH of natural enzymes and recombinant enzyme enzymatic reaction similar effect. Natural enzymes and recombinant enzyme were the highest priority cefoperazone substrate, followed by cephalexin; to ampicillin, amoxicillin, penicillin and piperacillin have the highest catalytic efficiency. Conclusion of natural enzymes and recombinant enzyme kinetic parameters was no significant difference. Keywords: β-lactamase kinetics of the recombinant protein β-lactam antibiotics Kinetic characteristics of native and recombinant TEM  116 beta  lactamases ABSTRACT Objective To investigate and compare the kinetic characteristics of native and recombinant TEM  116 beta  lactamases. Methods Hydrolysis reactions of beta  lactam antibiotics were detected by ultraviolet photometry. Data was processed according to Lee and Wilson’s improved double  reciprocal equation. Kinetic parameters of native and recombinant TEM  116 beta  lactamases, including Km, Vmax and kcat, were determined. Effects of temperature and pH on enzymatic reaction were observed.