Hybrid proteins with organophosphorus hydrolase activity and fluorescence of deGFP4 protein

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

Research output: Contribution to journalArticlepeer-review

Abstract

New genetic constructs encoding synthesis of hybrid proteins possessing organophosphorus hydrolase activity and properties of the pH sensitive analog of the green fluorescent protein were developed. It was established that 0.1 mM of the biosynthesis inducer and cultivation for 10 h after the induction were necessary for the maximum yield of the hybrid proteins in the soluble and active form in E. coli cells. The demonstrated synthesis level for one of the new proteins was 2- to 25-fold higher than the yield of the soluble hybrid protein analogs known from the literature. It was found that the organophosphorus hydrolase within hybrid proteins demonstrated characteristics (pH optimum, thermal stability and catalytic efficiency) different from the respective characteristics known for the native enzyme. It was shown that the fluorescence of the green fluorescent protein within the hybrid proteins depended on the pH of the medium in a manner similar to the individual protein. Interrelation of the fluorescent characteristics and OPH activity that manifested itself in the hydrolysis of organophosphorus compounds was shown by example of one of the hybrid proteins.

Original languageEnglish (US)
Pages (from-to)92-98
Number of pages7
JournalMoscow University Chemistry Bulletin
Volume66
Issue number2
DOIs
StatePublished - Apr 1 2011

Keywords

  • Green fluorescent protein
  • Hybrid proteins
  • Organophosphorus compounds
  • Organophosphorus hydrolase

ASJC Scopus subject areas

  • Chemistry(all)

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