ChemNet
 
Previous article Next article Contents  

D. A. Gudkov, I. V. Lyagin, V. V. Verkhusha, Ye. N. Efremenko

New chimeric proteins possessing organophosphoroushydrolase activity and deGFP4 fluorescence

Abstract

New genetic constructs encoding synthesis of chimeric proteins possessing organophosphoroushydrolase activity and fluorescence of pH-sensitive analogue of green fluorescent protein were developed. To obtain maximal yield of new chimeric proteins in soluble form in E. coli cells, the following conditions were revealed to be used: 0.1 mM of inductor of protein biosynthesis, cultivation of cells for 10 h after biosynthesis induction. Obtained level of biosynthesis of one of the chimeric proteins in soluble active form was 2-25-fold higher than the yields of soluble forms of analogue chimeric proteins described in literature. It was found that organophosphoroushydrolase as a part of the chimeric proteins possess features (pH-optimum, thermostability, substrate specificity and catalytic efficiency of action) differing from known characteristics of native enzyme. The fluorescence of green fluorescent protein being part of chimeric proteins depends on pH of medium by same manner known for fluorescence of the individual protein. The interrelation between fluorescence and OPH-activity possessing by obtained chimeric proteins was demonstrated in the reactions of hydrolysis of organophosphorous compounds.
Moscow University Chemistry Bulletin.
2011, Vol. 52, No. 2, P. 113
   

Copyright (C) Chemistry Dept., Moscow State University, 2002
   Overview
   Editorial board
   Tables of Contents
   Subscription

The site is supported by Russian Foundation for Basic Research
  The using of published on this page materials is not allowed without special permission
Copyright (C) Chemisty Department of Moscow State University
Web-Editor: B.I.Pokrovskii
Web-design: Copyright (C) MIG and VVM
webmaster@www.chem.msu.su