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.
Kriek
Marco
M
Department of Chemistry, Southampton University, Highfield, Southampton SO17 1BJ, UK.
Peters
Liv
L
Takahashi
Yasahiro
Y
Roach
Peter L
PL
D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D007506:Iron-Sulfur Proteins / Q000235:genetics / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D013466:Sulfurtransferases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D007506:Iron-Sulfur Proteins; D013466:Sulfurtransferases; C116435:lipoic acid synthase