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Article

Structural and Functional Characterization of New SsoPox Variant Points to the Dimer Interface as a Driver for the Increase in Promiscuous Paraoxonase Activity

1
Institute of Biochemistry and Cell Biology, CNR, 80131 Naples, Italy
2
The Weizmann Institute of Science, Rehovot, 76100 Israel
3
Institute of Biostructures and Bioimages, CNR, 80131 Naples, Italy
4
Bundeswehr Institute of Pharmacology and Toxicology, Munich, Germany
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(5), 1683; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21051683
Received: 17 December 2019 / Revised: 2 February 2020 / Accepted: 25 February 2020 / Published: 1 March 2020
(This article belongs to the Special Issue Thermophilic and Hyperthermophilic Microbes and Enzymes)
Increasing attention is more and more directed toward the thermostable Phosphotriesterase-Like-Lactonase (PLL) family of enzymes, for the efficient and reliable decontamination of toxic nerve agents. In the present study, the DNA Staggered Extension Process (StEP) technique was utilized to obtain new variants of PLL enzymes. Divergent homologous genes encoding PLL enzymes were utilized as templates for gene recombination and yielded a new variant of SsoPox from Saccharolobus solfataricus. The new mutant, V82L/C258L/I261F/W263A (4Mut) exhibited catalytic efficiency of 1.6 × 105 M−1 s−1 against paraoxon hydrolysis at 70°C, which is more than 3.5-fold and 42-fold improved in comparison with C258L/I261F/W263A (3Mut) and wild type SsoPox, respectively. 4Mut was also tested with chemical warfare nerve agents including tabun, sarin, soman, cyclosarin and VX. In particular, 4Mut showed about 10-fold enhancement in the hydrolysis of tabun and soman with respect to 3Mut. The crystal structure of 4Mut has been solved at the resolution of 2.8 Å. We propose that, reorganization of dimer conformation that led to increased central groove volume and dimer flexibility could be the major determinant for the improvement in hydrolytic activity in the 4Mut. View Full-Text
Keywords: Molecular evolution; Phosphotriesterase-Like-Lactonases (PLLs); organophosphate (OP) compounds; nerve agents; enzymatic detoxification/remediation Molecular evolution; Phosphotriesterase-Like-Lactonases (PLLs); organophosphate (OP) compounds; nerve agents; enzymatic detoxification/remediation
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MDPI and ACS Style

Suzumoto, Y.; Dym, O.; Roviello, G.N.; Worek, F.; Sussman, J.L.; Manco, G. Structural and Functional Characterization of New SsoPox Variant Points to the Dimer Interface as a Driver for the Increase in Promiscuous Paraoxonase Activity. Int. J. Mol. Sci. 2020, 21, 1683. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21051683

AMA Style

Suzumoto Y, Dym O, Roviello GN, Worek F, Sussman JL, Manco G. Structural and Functional Characterization of New SsoPox Variant Points to the Dimer Interface as a Driver for the Increase in Promiscuous Paraoxonase Activity. International Journal of Molecular Sciences. 2020; 21(5):1683. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21051683

Chicago/Turabian Style

Suzumoto, Yoko, Orly Dym, Giovanni N. Roviello, Franz Worek, Joel L. Sussman, and Giuseppe Manco. 2020. "Structural and Functional Characterization of New SsoPox Variant Points to the Dimer Interface as a Driver for the Increase in Promiscuous Paraoxonase Activity" International Journal of Molecular Sciences 21, no. 5: 1683. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms21051683

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