Abstract
Thermostable phosphotriesterase-like lactonases (PLLs) from extremophile archaea, like SsoPox from Sulfolobus solfataricus, are attractive biotechnological tools with industrial applications as organophosphate decontaminants, but their manufacturing still remains an unresolved issue because of the high costs and the low production yields. In this paper, for the first time, an efficient biotechnological process for the production and purification of a recombinant, engineered PLL, SsoPox W263F, expressed in E. coli, has been set up by studying new induction strategies, by designing high cell density cultivations and a new membrane-based downstream process. In fed batches, the enzyme production was boosted of 69-fold up to 4660.0 U L−1 using galactose as inducer in the replacement of IPTG; the process was scalable from 2.5 up to 150 L. By coupling a single thermo-precipitation step and an ultrafiltration process, a total enzyme recovery of 77% with a purity grade of almost 80% was reached.
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Acknowledgements
This work was supported by the Italian Ministry of the University and Research (MIUR) and by the European Union Commission (project PON_01585 to Giuseppe Manco). All the authors declare that they have no conflict of interest.
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Restaino, O.F., Borzacchiello, M.G., Scognamiglio, I. et al. Boosted large-scale production and purification of a thermostable archaeal phosphotriesterase-like lactonase for organophosphate decontamination. J Ind Microbiol Biotechnol 44, 363–375 (2017). https://doi.org/10.1007/s10295-016-1892-x
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DOI: https://doi.org/10.1007/s10295-016-1892-x