Characterization of the Dihydroorotase from Methanococcus jannaschii.

ABSTRACT: The gene that codes for the putative dihydroorotase in the hyperthermophilic archaeon Methanococcus jannaschii was subcloned in pET-21a and expressed in Escherichia coli. A purification protocol was devised. The purity of the protein was evaluated by SDS-PAGE and the protein was confirmed by sequencing using LC-MS. The calculated molecular mass is 48104 Da. SEC-LS suggested that the protein is a monomer in solution. ICP-MS showed that there are two Zn ions per monomer. Kinetic analysis of the recombinant protein gave hyperbolic kinetics with V_max = 12.2 µmol/min/mg and K_m = 0.14 mM at 25 °C. Furthermore the activity of the protein increased with temperature consistent with the hyperthermophilic nature of the organism. A homology model was constructed using the mesophi

SUBMITTER: Vitali J 

PROVIDER: S-EPMC5624517 | biostudies-literature | 2017 Aug

REPOSITORIES: biostudies-literature