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Appl. Sci. 2017, 7(12), 1305; doi:10.3390/app7121305

Intensification of Organophosphorus Hydrolase Synthesis by Using Substances with Gas-Transport Function

1
Faculty of Chemistry, Lomonosov Moscow State University, Moscow 119991, Russia
2
A. N. Nesmeyanov Institute of Organoelement Compounds of Russian Academy of Sciences, Moscow 119991, Russia
3
A. N. Frumkin Institute of Physical Chemistry and Electrochemistry of Russian Academy of Sciences, Moscow 119071, Russia
*
Author to whom correspondence should be addressed.
Received: 3 November 2017 / Revised: 6 December 2017 / Accepted: 13 December 2017 / Published: 17 December 2017
(This article belongs to the Section Chemistry)
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Abstract

We have performed studies and comparative analysis of the biosynthesis characteristics of intracellular recombinant enzyme, such as hexahistidine-containing organophosphorus hydrolase (His6-OPH) in Escherichia coli SG13009[pREP4] cells when various perfluorocarbon compounds (PFC) were introduced into the medium for cell cultivation. The PFC were found to facilitate the biosynthesis of His6-OPH: increased levels of the total OPH-activity (up to 37%) were measured upon introduction of 1,1,1,2,2,3,3,4,4,5,5,6,6,6-tetradecafluorohexane (PFH) and 4,7,10,13,16,19,22,25,28,31-decaoxaperfluoro-5,8,11,14,17,18,21,24,27,30-decamethyl tetratriacontane (Polyether II) into culture medium. We have demonstrated the possibility of effective and multiple (at least five-fold) use of PFH for biosynthesis of intracellular recombinant protein His6-OPH, which catalyzes the hydrolysis of organophosphorus pesticides (OP), is widely used in agriculture and can be applied as new antidote for OP-detoxification in vivo. The multiple use of PFH was achieved through recycling of this substance: sediment of Escherichia coli SG13009[pREP4] cell biomass was collected at the end of each culture growing step and disintegrated with ultrasound, and obtained residue containing almost all of the initially introduced PFC was then added to the medium at the start of the following culture growing step. View Full-Text
Keywords: perfluorocarbon compounds; perfluorohexane; recombinant intracellular protein; hexahisdine-containing organophosphorus hydrolase; Escherichia coli perfluorocarbon compounds; perfluorohexane; recombinant intracellular protein; hexahisdine-containing organophosphorus hydrolase; Escherichia coli
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Senko, O.; Stepanov, N.; Tyutyunov, A.; Sterlin, S.; Grinberg, V.; Makhlis, T.; Efremenko, E. Intensification of Organophosphorus Hydrolase Synthesis by Using Substances with Gas-Transport Function. Appl. Sci. 2017, 7, 1305.

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