N-terminal deletion of Tk1689, a subtilisin-like serine protease from Thermococcus kodakaraensis, copes with its cytotoxicity in Escherichia coli

Nouman Rasool; Naeem Rashid; Saima Iftikhar; Muhammad Akhtar

doi:10.1016/j.jbiosc.2010.04.005

N-terminal deletion of Tk1689, a subtilisin-like serine protease from Thermococcus kodakaraensis, copes with its cytotoxicity in Escherichia coli

J Biosci Bioeng. 2010 Oct;110(4):381-5. doi: 10.1016/j.jbiosc.2010.04.005. Epub 2010 May 8.

Authors

Nouman Rasool¹, Naeem Rashid, Saima Iftikhar, Muhammad Akhtar

Affiliation

¹ School of Biological Sciences, University of the Punjab, Lahore 54570, Pakistan.

PMID: 20547373
DOI: 10.1016/j.jbiosc.2010.04.005

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.

MeSH terms

Base Sequence
Biocatalysis
DNA Primers
Electrophoresis, Polyacrylamide Gel
Enzyme Stability
Escherichia coli / cytology*
Hot Temperature
Hydrogen-Ion Concentration
Models, Molecular
Recombinant Proteins / chemistry
Recombinant Proteins / metabolism
Subtilisins / chemistry
Subtilisins / metabolism*
Thermococcus / enzymology*

Substances

DNA Primers
Recombinant Proteins
Subtilisins