Molecular cloning, purification, and biochemical characterization of hydantoin racemase from the legume symbiont Sinorhizobium meliloti CECT 4114.
Hydantoin racemase from Sinorhizobium meliloti was functionally expressed in Escherichia coli. The native form of the enzyme was a homotetramer with a molecular mass of 100 kDa. The optimum temperature and pH for the enzyme were 40 degrees C and 8.5, respectively. The enzyme showed a slight preference for hydantoins with short rather than long aliphatic side chains or those with aromatic rings. Substrates, which showed no detectable activity toward the enzyme, were found to exhibit competitive inhibition.
Martínez-Rodríguez
Sergio
S
Departamento de Química-Física, Bioquímica y Química Inorgánica, Universidad de Almería, La Cañada de San Urbano, E-04120 Almería, Spain.
Las Heras-Vázquez
Francisco Javier
FJ
Mingorance-Cazorla
Lydia
L
Clemente-Jiménez
Josefa María
JM
Rodríguez-Vico
Felipe
F
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D007887:Fabaceae / Q000382:microbiology*; D006827:Hydantoins / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D019998:Racemases and Epimerases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016415:Sequence Alignment; D016962:Sinorhizobium meliloti / Q000201:enzymology*; D013237:Stereoisomerism; D013379:Substrate Specificity; D013559:Symbiosis
D006827:Hydantoins; D019998:Racemases and Epimerases; C074773:hydantoin racemase