High level secretion of TaqI restriction endonuclease by recombinant Escherichia coli

E. Toksoy; P. H. Özdinler; Z. I. Önsan; B. Kirdar

doi:10.1023/A:1008938302312

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

Neurology

Research output: Contribution to journal › Article › peer-review

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 language	English (US)
Pages (from-to)	803-808
Number of pages	6
Journal	Biotechnology Techniques
Volume	13
Issue number	11
DOIs	https://doi.org/10.1023/A:1008938302312
State	Published - 1999

Keywords

Extracellular production
MBP fusion protein
Secretion
TaqI restriction endonuclease

ASJC Scopus subject areas

Biochemistry
Applied Microbiology and Biotechnology

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10.1023/A:1008938302312

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title = "High level secretion of TaqI restriction endonuclease by recombinant Escherichia coli",

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.",

keywords = "Extracellular production, MBP fusion protein, Secretion, TaqI restriction endonuclease",

author = "E. Toksoy and {\"O}zdinler, {P. H.} and {\"O}nsan, {Z. I.} and B. Kirdar",

note = "Funding Information: The financial support for this research was provided by TUBITAK Research Council TBAG-1533 and Bogazici University Research Fund Projects 96A0522, 97A0501 and DPT-95K120320.",

year = "1999",

doi = "10.1023/A:1008938302312",

language = "English (US)",

volume = "13",

pages = "803--808",

journal = "Biotechnology Techniques",

issn = "0951-208X",

publisher = "Springer Netherlands",

number = "11",

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TY - JOUR

T1 - High level secretion of TaqI restriction endonuclease by recombinant Escherichia coli

AU - Toksoy, E.

AU - Özdinler, P. H.

AU - Önsan, Z. I.

AU - Kirdar, B.

N1 - Funding Information: The financial support for this research was provided by TUBITAK Research Council TBAG-1533 and Bogazici University Research Fund Projects 96A0522, 97A0501 and DPT-95K120320.

PY - 1999

Y1 - 1999

N2 - 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.

AB - 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.

KW - Extracellular production

KW - MBP fusion protein

KW - Secretion

KW - TaqI restriction endonuclease

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DO - 10.1023/A:1008938302312

M3 - Article

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SN - 0951-208X

VL - 13

SP - 803

EP - 808

JO - Biotechnology Techniques

JF - Biotechnology Techniques

IS - 11

ER -

High level secretion of TaqI restriction endonuclease by recombinant Escherichia coli

Abstract

Keywords

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