Effect of iron-sulfur cluster assembly proteins on the expression of Escherichia coli lipoic acid synthase
Effect of iron-sulfur cluster assembly proteins on the expression of Escherichia coli lipoic acid synthase
Lipoic Acid Synthase (LipA) can accommodate a [4Fe-4S] cluster that is thought to be essential for the insertion of sulfur into an octanoyl substrate during the biosynthesis of lipoic acid. With the objective of improving soluble holo-LipA expression, a series of multi-cistronic plasmids were constructed carrying lipA in combination with one of the three systems: groE/SL, trxA, or the isc operon. Co-expression of lipA with the isc operon approximately trebled the isolated yield of soluble LipA and resulted in efficient assembly of the Fe-S cluster. This strategy may be helpful in the soluble expression of a wide range of Fe-S cluster-dependent proteins.
lipoate synthase, gene-cluster, lipa, biosynthesis, complex
241-245
Kriek, Marco
c3e820c2-c59b-49c8-a792-f64a69e4401a
Peters, Liv
c88eab5b-ef1e-41a5-adc5-bcd40f1e4a2f
Takahashi, Yasahiro
5c8696ca-cfa4-47ab-9fb6-6f2038499fca
Roach, Peter L.
ca94060c-4443-482b-af3e-979243488ba9
1 April 2003
Kriek, Marco
c3e820c2-c59b-49c8-a792-f64a69e4401a
Peters, Liv
c88eab5b-ef1e-41a5-adc5-bcd40f1e4a2f
Takahashi, Yasahiro
5c8696ca-cfa4-47ab-9fb6-6f2038499fca
Roach, Peter L.
ca94060c-4443-482b-af3e-979243488ba9
Kriek, Marco, Peters, Liv, Takahashi, Yasahiro and Roach, Peter L.
(2003)
Effect of iron-sulfur cluster assembly proteins on the expression of Escherichia coli lipoic acid synthase.
Protein Expression and Purification, 28 (2), .
(doi:10.1016/S1046-5928(02)00680-0).
Abstract
Lipoic Acid Synthase (LipA) can accommodate a [4Fe-4S] cluster that is thought to be essential for the insertion of sulfur into an octanoyl substrate during the biosynthesis of lipoic acid. With the objective of improving soluble holo-LipA expression, a series of multi-cistronic plasmids were constructed carrying lipA in combination with one of the three systems: groE/SL, trxA, or the isc operon. Co-expression of lipA with the isc operon approximately trebled the isolated yield of soluble LipA and resulted in efficient assembly of the Fe-S cluster. This strategy may be helpful in the soluble expression of a wide range of Fe-S cluster-dependent proteins.
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Published date: 1 April 2003
Keywords:
lipoate synthase, gene-cluster, lipa, biosynthesis, complex
Identifiers
Local EPrints ID: 20002
URI: http://eprints.soton.ac.uk/id/eprint/20002
ISSN: 1046-5928
PURE UUID: 34d4a4dd-5155-49e5-af43-e1d60f552810
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Date deposited: 24 Feb 2006
Last modified: 15 Mar 2024 06:21
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Contributors
Author:
Marco Kriek
Author:
Liv Peters
Author:
Yasahiro Takahashi
Author:
Peter L. Roach
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