title: N-terminal deletion of Tk1689, a subtilisin-like serine protease from Thermococcus kodakaraensis, copes with its cytotoxicity in Escherichia coli.
abstract: Tk1689, a subtilisin-like serine protease from Thermococcus kodakaraensis, was found to be toxic to the host cells when produced in the pro-protein form (Pro-Tk1689) in Eschericia coli. Cytotoxic effect of Pro-Tk1689 was reduced when signal and pro-peptide both were removed and the protein was produced in the mature form (MP-Tk1689). The mature protein was produced in E. coli in the soluble form. Recombinant MP-Tk1689 was catalytically active and exhibited optimum activity at 55°C and pH 7. Specific activity of the enzyme was 700 U/mg. The enzyme displayed a half life of 80 min at 60°C.
authors.affiliation: School of Biological Sciences, University of the Punjab, Lahore 54570, Pakistan.
authors.forename: Nouman
authors.initials: N
authors.lastname: Rasool
authors.forename: Naeem
authors.initials: N
authors.lastname: Rashid
authors.forename: Saima
authors.initials: S
authors.lastname: Iftikhar
authors.forename: Muhammad
authors.initials: M
authors.lastname: Akhtar
mesh_terms: D001483:Base Sequence; D055162:Biocatalysis; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D004926:Escherichia coli / Q000166:cytology*; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013381:Subtilisins / Q000737:chemistry / Q000378:metabolism*; D019712:Thermococcus / Q000201:enzymology*
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D013381:Subtilisins