Cloning and expression of the glutamate racemase gene of Bacillus pumilus.
A glutamate racemase gene (murI) was found in Bacillus pumilus cells and cloned into Escherichia coli WM335, a D-glutamate auxotroph, by means of a genetic complement method. MurI of B. pumilus encodes a 272-amino acid protein with an unusual initiation codon, TTG. The deduced amino acid sequence shows significant similarity with those of glutamate racemases from E. coli (ratio of identical residues, 28%), Pediococcus pentosaceus (44%), and Staphylococcus haemolyticus (49%). B. pumilus MurI was expressed as a fusion protein connected to the N-terminal 12 residues of beta-galactosidase; the fusion protein showed glutamate racemase activity, and resembled the enzyme of P. pentosaceus in physicochemical and enzymological properties.
Liu
L
L
Institute for Chemical Research, Kyoto University, Uji.
Yoshimura
T
T
Endo
K
K
Esaki
N
N
Soda
K
K
D000591:Amino Acid Isomerases / Q000235:genetics*; D000595:Amino Acid Sequence; D001407:Bacillus / Q000235:genetics*; D001412:Bacillus subtilis; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D004926:Escherichia coli; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D008969:Molecular Sequence Data; D009154:Mutation; D010442:Peptide Chain Initiation, Translational / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis
D003062:Codon; D011993:Recombinant Fusion Proteins; D000591:Amino Acid Isomerases; C062292:glutamate racemase