Gene for aspartate racemase from the sulfur-dependent hyperthermophilic archaeum, Desulfurococcus strain SY.
Amino acid racemases are ubiquitous throughout eubacteria. However, no amino acid racemases have yet been found in eukaryotes and archaea. We cloned a gene highly homologous to that for the aspartate racemase from the sulfur-dependent hyperthermophilic archaeum, Desulfurococcus strain SY. The product of the gene showed 35.2% amino acid sequence identity with the aspartate racemase of Streptococcus thermophilus IAM10064, and was also homologous to glutamate racemases around the putative catalytic cysteine residues. The encoded protein was expressed in Escherichia coli. The recombinant protein had amino acid racemizing activity, which was highly specific for aspartate and increased with temperature from 37 degrees C to 90 degrees C. Therefore, this was identified as the first hyperthermophilic archaeal amino acid racemase. A little aspartate racemizing activity was also detected in the crude extract of Desulfurococcus strain SY. The function of this aspartate racemase might be the uptake of -aspartate formed at high temperature or the production of -aspartate as a cell component. The fact that the amino acid racemases are distributed among both eubacteria and archaea suggests that endogenous -amino acids in mammals are also synthesized by amino acid racemases.
Yohda
M
M
Biochemical Systems Laboratory, The Institute of Physical and Chemical Research, Wako, Saitama 351-01.
Endo
I
I
Abe
Y
Y
Ohta
T
T
Iida
T
T
Maruyama
T
T
Kagawa
Y
Y
D000591:Amino Acid Isomerases / Q000235:genetics*; D000595:Amino Acid Sequence; D001105:Archaea / Q000201:enzymology*; D001483:Base Sequence; D004269:DNA, Bacterial; D004795:Enzyme Stability; D005784:Gene Amplification; D006358:Hot Temperature; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D000591:Amino Acid Isomerases; C019749:aspartate racemase