The cloning, expression and crystallisation of a thermostable arginase

Maria C. Bewley; J. Shaun Lott; Edward N. Baker; Mark L. Patchett

doi:10.1016/0014-5793(96)00459-0

The cloning, expression and crystallisation of a thermostable arginase

Maria C. Bewley, J. Shaun Lott, Edward N. Baker, Mark L. Patchett

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Abstract

The gene for the thermostable arginase from the thermophilic bacterium 'Bacillus caldovelox' has been cloned and sequenced. Expression of recombinant arginase at high levels has been achieved in E. coli using an inducible T7 RNA polymerase-based system. A facile purification procedure incorporating a heat-treatment step yielded 0.2 g of recombinant arginase per litre of induced culture. The kinetic properties of the purified recombinant protein are essentially identical to the native enzyme. The recombinant protein has been crystallised and one crystal form is isomorphous to crystals of the native protein.

Original language	English (US)
Pages (from-to)	215-218
Number of pages	4
Journal	FEBS Letters
Volume	386
Issue number	2-3
DOIs	https://doi.org/10.1016/0014-5793(96)00459-0
State	Published - May 20 1996

All Science Journal Classification (ASJC) codes

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/0014-5793(96)00459-0

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title = "The cloning, expression and crystallisation of a thermostable arginase",

abstract = "The gene for the thermostable arginase from the thermophilic bacterium 'Bacillus caldovelox' has been cloned and sequenced. Expression of recombinant arginase at high levels has been achieved in E. coli using an inducible T7 RNA polymerase-based system. A facile purification procedure incorporating a heat-treatment step yielded 0.2 g of recombinant arginase per litre of induced culture. The kinetic properties of the purified recombinant protein are essentially identical to the native enzyme. The recombinant protein has been crystallised and one crystal form is isomorphous to crystals of the native protein.",

author = "Bewley, {Maria C.} and Lott, {J. Shaun} and Baker, {Edward N.} and Patchett, {Mark L.}",

note = "Funding Information: Acknowledgements: We gratefully acknowledge financial support from the Foundation for Research, Science and Technology. E.N.B. also receives support as an International Research Scholar of the Howard Hughes Medical Institute.",

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T1 - The cloning, expression and crystallisation of a thermostable arginase

AU - Bewley, Maria C.

AU - Lott, J. Shaun

AU - Baker, Edward N.

AU - Patchett, Mark L.

N1 - Funding Information: Acknowledgements: We gratefully acknowledge financial support from the Foundation for Research, Science and Technology. E.N.B. also receives support as an International Research Scholar of the Howard Hughes Medical Institute.

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The cloning, expression and crystallisation of a thermostable arginase

Abstract

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