• 한국군사과학기술학회지
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• 제2권2호
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• pp.140-147
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• 1999

A gene expressing organophosphorus acid (OPA) anhydrolases have been cloned from Alteromonas haloplanktis strain and expressed in bacterial strain BL21. Crude extract was prepared from the transformed bacterial strain BL2l and used in testing its degrading capability of real nerve gas, soman and sarin. Within 1 minute after the start of the reaction, nearly 65% of the soman added to the reactant(3mM) was degraded by adding 1 mg of the crude extract enzyme(20.0 Unit $mg^-{1}$ crude protein). In 6 minutes, the reaction reached at its steady state, which indicates that soman was completely degraded by that time. In the case of sarin, the degradation efficiency was observed to be about 0.7 times of that of soman. If the specific activity of OPAA is enhanced by both increased expression efficiency and purification, OPAA seems to be applied for the development of decontaminant of skin, especially of eye.

• 한국군사과학기술학회지
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• 제4권1호
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• pp.188-197
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• 2001

Organophosphorus acid anhydrolases(OPAA) catalyzing the hydrolysis of toxic organophosphates have been found in a variety of prokaryotic and eukaryotic organisms. Of the several kinds of OPAA that can degrade nerve agents, such as DFP, sarin and soman, a OPAA gene harbored in the chromosomal DNA of Alteromonas haloplanktis strain was subcloned in order to develope an enzymatic degradation method of toxic organophosphorus compounds. For this 1481 bp DNA fragment containing OPAA gene and its flanking regions has been synthesized through PCR using chromosomal DNA of A. haloplanktis strain. After subcloning and subsequent expression, crude OPA anhydrolase was prepared and assayed. It was shown that the OPAA had a very high hydrolytic activity on DFP. The specific activity of the enzyme was 1,110 $\mu$mole.$min^{p-1}.mg^{-1}$ protein. It seemed that OPAA with such a high hydrolytic activity may give a good prospects to its use, as a biodegradation tool, in detoxifying toxic organophosphorus compounds, such as pesticides and chemical stockpiles which are posing a potential threat to the field environment and human health.