title: Improving Properties of Recombinant SsoPox by Site-Specific Pegylation.
abstract: SsoPox, a ~35 kDa enzyme from Sulfolobus solfataricus, can hydrolyze and inactivate a variety of organophosphate (OP)-compounds. The enzyme is a potential candidate for the development of prophylactic and therapeutic agent against OP-poisoning in humans. However, the therapeutic use of recombinant SsoPox suffers from certain limitations associated with the use of recombinant protein pharmaceuticals. Some of these limitations could be overcome by conjugating SsoPox enzyme with polyethylene glycol (PEG). In this study, we report generation and in vitro characterization of N-terminal mono-PEGylated rSsoPox(2p) (a variant of rSsoPox(wt) having enhanced OP-hydrolyzing activity). The enzyme was PEGylated with mPEG-propionaldehyde and the PEGylated protein was isolated using ion-exchange chromatography. Compared with the unmodified enzyme, mono-PEGylation of rSsoPox results in improvement in the thermostability and protease resistance of the enzyme. PEGylated rSsoPox(2p) can be developed as a candidate for the prevention / treatment of OP-poisoning.
authors.forename: Harsh
authors.initials: H
authors.lastname: Parikh
authors.forename: Priyanka
authors.initials: P
authors.lastname: Bajaj
authors.forename: Rajan K
authors.initials: RK
authors.lastname: Tripathy
authors.affiliation: Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), Sector 67, S.A.S. Nagar, (Mohali)-160 062, Punjab, India. abbupande@yahoo.co.in.
authors.forename: Abhay H
authors.initials: AH
authors.lastname: Pande
mesh_terms: D000581:Amidohydrolases / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D004795:Enzyme Stability; D044345:Phosphoric Triester Hydrolases / Q000737:chemistry* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism; D048229:Sulfolobus solfataricus / Q000201:enzymology*; D014357:Trypsin
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D044345:Phosphoric Triester Hydrolases; D014357:Trypsin; D000581:Amidohydrolases