High level secretion of TaqI restriction endonuclease by recombinant Escherichia coli

E. Toksoy, P. H. Özdinler, Z. I. Önsan, B. Kirdar*

*Corresponding author for this work

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

The production and high level secretion of TaqI restriction endonuclease using bacterial secretion signal within the male gene was achieved by cloning the PCR-amplified gene from Thermus aquaticus into E. coli. The maltose binding protein (MBP) part of the MBP-TaqI fusion protein expressed by this construct did not interfere with the biological activity of the TaqI restriction endonuclease. E. coli XL1 carrying pil185 produced 332 U ml-1 TaqI endonuclease 81% of which was secreted into the medium without apparent cell lysis. Optimization of culture conditions and selection of the host strain were found to be important for the efficient extracellular production of this protein.

Original languageEnglish (US)
Pages (from-to)803-808
Number of pages6
JournalBiotechnology Techniques
Volume13
Issue number11
DOIs
StatePublished - Jan 1 1999

Keywords

  • Extracellular production
  • MBP fusion protein
  • Secretion
  • TaqI restriction endonuclease

ASJC Scopus subject areas

  • Biochemistry
  • Applied Microbiology and Biotechnology

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