High level secretion of TaqI restriction endonuclease by recombinant Escherichia coli

Toksoy E., Ozdinler P., Onsan Z., Kirdar B.

BIOTECHNOLOGY TECHNIQUES, cilt.13, sa.11, ss.803-808, 1999 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 13 Sayı: 11
  • Basım Tarihi: 1999
  • Doi Numarası: 10.1023/a:1008938302312
  • Dergi Adı: BIOTECHNOLOGY TECHNIQUES
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.803-808
  • Anahtar Kelimeler: TaqI restriction endonuclease, MBP fusion protein, secretion, extracellular production, BACTERIOCIN-RELEASE-PROTEIN, EXTRACELLULAR PRODUCTION, EXPRESSION, SYSTEM, STREPTOKINASE, EXCRETION
  • Marmara Üniversitesi Adresli: Hayır

Özet

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 pH185 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.