Hydantoin racemase from Arthrobacter aurescens DSM 3747: heterologous expression, purification and characterization

Molecular Cloning and Expression of thehyuGenes fromMicrobacterium liquefaciensAJ 3912, Responsible for the Conversion of 5-Substituted Hydantoins to α-Amino Acids, inEscherichia coli

“…The estimated molecular mass (25,251 Da) agreed well with the apparent molecular mass of MHR as determined by SDS-PAGE in our previous study (27 kDa), 11) and was similar to those of known HRase proteins (25-27 kDa). [6][7][8][9][10] The deduced amino acid sequence of MHR showed homology with those of other HRase proteins (Fig. 1), particularly the HRase from A. aurescens DSM 3747 (AaHyuA).…”

“…The literature on AaHyuA notes the contribution of its cysteine residues to the expression of HRase activity and the existence of two highly conserved cysteine residues in the sequences of HRase proteins in general. 7) In the case of MHR, HRase activity was inhibited by cysteinemodifying reagents, 11) and the molecular subunit contains three cysteine residues at positions 77, 182, and 208. The cysteine residues at positions 77 and 182 were highly conserved among HRase proteins (Fig.…”

“…NS761, Arthrobacter aurescens DSM 3747, Agrobacterium tumefaciens C58, and Sinorhizobium meliloti CECT 4114. [6][7][8][9][10] Recently, we also isolated a new HRase protein from Microbacterium liquefaciens AJ 3912 (MHR). 11) This strain was first isolated as a potent producer of L-amino acids from the corresponding DL-5-substituted-hydantoins, and is also known to possess both a D,L-nonstereoselective hydantoinase (MHH) and a L-stereoselective N-carbamoylase (MCH).…”

Molecular Cloning, Purification, and Biochemical Characterization of Hydantoin Racemase from the Legume Symbiont Sinorhizobium meliloti CECT 4114

“…2) was very similar to those of Pseudomonas sp. strain NS671 (32 kDa), Arthrobacter aurescens DSM 3747 (31 kDa), and Agrobacterium tumefaciens C58 (31 kDa) (8,20,21). In all cases, these apparent molecular masses were greater than those calculated from the amino acid sequence (25 to 27 kDa).…”

“…Likewise, the hydantoin racemase enzyme involved in the production of L-amino acids from Pseudomonas sp. strain NS671 (20) and Arthrobacter aurescens DSM 3747 (21), and more recently the one involved in the production of D-amino acids from Agrobacterium tumefaciens (8,10), have been purified and biochemically characterized. Sinorhizobium meliloti is an ␣-proteobacterium of the family Rhizobiaceae, as is Agrobacterium tumefaciens, which forms agronomically important N 2 -fixing root nodules in legumes (4).…”