A novel enzyme, D-3-hydroxyaspartate aldolase from Paracoccus denitrificans IFO 13301: purification, characterization, and gene cloning.
A novel enzyme, D-3-hydroxyaspartate aldolase (D-HAA), catalyzing the conversion of D-3-hydroxyaspartate to glyoxylate plus glycine, was purified to homogeneity from Paracoccus denitrificans IFO 13301. D-HAA is strictly D-specific as to the alpha-position, whereas the enzyme does not distinguish between threo and erythro forms at the beta-position. In addition to D-3-hydroxyaspartate, the enzyme also acts on d-threonine, D-3-3,4-dihydroxyphenylserine, D-3-3,4-methylenedioxyphenylserine, and D-3-phenylserine. The D-HAA gene was cloned and sequenced. The gene contains an open reading frame consisting of 1,161 nucleotides corresponding to 387 amino acid residues. The predicted amino acid sequence displayed 35% and 22% identity with that of the D-threonine aldolase of Arthrobacter sp. DK-38 and Alcaligenes xylosoxidan IFO 12669, respectively. This is the first paper reporting both a purified enzyme with D-3-hydroxyaspartate aldolase activity and also its gene cloning.
Liu
J Q
JQ
Biotechnology Research Center, Toyama Prefectural University, Kurokawa 5180, 939-0398 Kosugi, Toyama, Japan.
Dairi
T
T
Itoh
N
N
Kataoka
M
M
Shimizu
S
S
D000595:Amino Acid Sequence; D001224:Aspartic Acid / Q000031:analogs & derivatives / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010231:Paracoccus denitrificans / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature
C011274:3-hydroxyaspartic acid; D001224:Aspartic Acid; D005634:Fructose-Bisphosphate Aldolase