Expression, purification and characterization of a cysteine desulfurase, IscS, from Acidithiobacillus ferrooxidans.
Iron-sulfur clusters are one of the most common types of redox center in nature. Three proteins of IscS (a cysteine desulfurase), IscU (a scaffold protein) and IscA (an iron chaperon) encoded by the operon iscSUA are involved in the iron-sulfur cluster assembly in Acidithiobacillus ferrooxidans. In this study the gene of IscS from A. ferrooxidans ATCC 23270 was cloned and expressed in Escherichia coli, the protein was purified by one-step affinity chromatography to homogeneity. The molecular mass of recombinant IscS was 46 kDa by SDS-PAGE. The IscS was a pyridoxal phosphate-containing protein, that catalyzed the elimination of S from L: -cysteine to yield L: -alanine and elemental sulfur or H(2)S, depending on whether or not a reducing agent was added to the reaction mixture.
Department of Bioengineering, School of Resources Processing and Bioengineering, Central South University, Changsha, 410083, PR China.
Jia
J
Zeng
Yanfei
Y
Zhang
Yuandong
Y
Liu
Xiaojian
X
Zhang
Leixian
L
Xia
Jianshe
J
Liu
Guanzhou
G
Qiu
D042763:Acidithiobacillus / Q000201:enzymology* / Q000235:genetics; D000595:Amino Acid Sequence; D001051:Apoenzymes / Q000378:metabolism; D013437:Carbon-Sulfur Lyases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D020035:Holoenzymes / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D013056:Spectrophotometry, Ultraviolet
D001051:Apoenzymes; D020035:Holoenzymes; D011994:Recombinant Proteins; D013437:Carbon-Sulfur Lyases; C080546:cysteine desulfurase