A novel metal-activated L-serine O-acetyltransferase from Thermus thermophilus HB8.
L-Cysteine is an important amino acid in terms of its industrial applications. The biosynthesis of L-cysteine in enteric bacteria is regulated through the feedback inhibition by L-cysteine of L-serine O-acetyltransferase (SAT), a key enzyme in L-cysteine biosynthesis. We recently found that L-cysteine is overproduced in Escherichia coli strains expressing a gene encoding feedback inhibition-insensitive SAT. Further improvements in L-cysteine production are expected by the use of SAT with high stability. We report here the sat1 gene encoding SAT of an extreme thermophile, Thermus thermophilus HB8. The sat1 gene was cloned and overexpressed in E. coli cells based on the genome sequence in T. thermophilus HB8. The predicted amino acid sequence consists of 295 amino acids and is homologous to other O-acetyltransferase members. In particular, the carboxyl-terminal region shares approximately 30% identities with SATs found in bacteria and plants, despite showing only about 15% identity in the overall sequence. Enzymatic analysis and an atomic absorption study of the purified recombinant proteins revealed that the enzyme is highly activated by Co(2+) or Ni(2+), and contains Zn(2+) and Fe(2+). These results indicate that the T. thermophilus SAT is a novel type of enzyme different from other members of this protein family.
Kobayashi
Shin-ichiro
S
RIKEN Harima Institute/Spring-8, Hyogo 679-5148, Japan.
Masui
Ryoji
R
Yokoyama
Shigeyuki
S
Kuramitsu
Seiki
S
Takagi
Hiroshi
H
D000123:Acetyltransferases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D003545:Cysteine / Q000096:biosynthesis; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D019216:Metals, Heavy / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D051050:Serine O-Acetyltransferase; D013045:Species Specificity; D013379:Substrate Specificity; D016963:Thermus thermophilus / Q000201:enzymology* / Q000235:genetics*
D019216:Metals, Heavy; D011994:Recombinant Proteins; D000123:Acetyltransferases; D051050:Serine O-Acetyltransferase; D003545:Cysteine