Boosted large-scale production and purification of a thermostable archaeal phosphotriesterase-like lactonase for organophosphate decontamination

Autor: Elena Porzio, Giuseppe Manco, Odile Francesca Restaino, Cinzia Donatiello, Chiara Schiraldi, Luigi Fedele, Mario De Rosa, Ilaria Scognamiglio, Maria Giovanna Borzacchiello
Přispěvatelé: Restaino, Odile Francesca, Borzacchiello, Maria Giovanna, Scognamiglio, Ilaria, Porzio, Elena, Manco, Giuseppe, Fedele, Luigi, Donatiello, Cinzia, Rosa, Mario De, Schiraldi, Chiara
Rok vydání: 2016
Předmět:
0106 biological sciences
0301 basic medicine
ved/biology.organism_classification_rank.species
Ultrafiltration
Bioengineering
Protein Engineering
01 natural sciences
Applied Microbiology and Biotechnology
Genes
Archaeal
03 medical and health sciences
chemistry.chemical_compound
Industrial Microbiology
010608 biotechnology
Lactonase
Escherichia coli
Extremophile
Decontamination
biology
ved/biology
Sulfolobus solfataricus
Organophosphate
High cell density cultivation Organophosphates Sulfolobus solfataricus Thermostable phosphotriesterase-like lactonase Ultrafiltration
High cell
Human decontamination
biology.organism_classification
Organophosphates
Culture Media
030104 developmental biology
Phosphoric Triester Hydrolases
Biochemistry
chemistry
Batch Cell Culture Techniques
Fermentation
biology.protein
Microorganisms
Genetically-Modified
Carboxylic Ester Hydrolases
Biotechnology
Archaea
Zdroj: Journal of industrial microbiology & biotechnology 44 (2017): 363–375. doi:10.1007/s10295-016-1892-x
info:cnr-pdr/source/autori:Restaino O.F.; Borzacchiello M.G.; Scognamiglio I.; Porzio E.; Manco G.; Fedele L.; Donatiello C.; De Rosa M.; Schiraldi C./titolo:Boosted large-scale production and purification of a thermostable archaeal phosphotriesterase-like lactonase for organophosphate decontamination/doi:10.1007%2Fs10295-016-1892-x/rivista:Journal of industrial microbiology & biotechnology/anno:2017/pagina_da:363/pagina_a:375/intervallo_pagine:363–375/volume:44
ISSN: 1476-5535
DOI: 10.1007/s10295-016-1892-x
Popis: 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.
Databáze: OpenAIRE
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