Microbial Conversion ofDL-5-Substituted Hydantoins to the CorrespondingL5-Amino Acids byBacillus stearothermophilusNS1122A

N-Carbamyl-L-Amino Acid Amidohydrolase ofPseudomonassp. Strain NS671: Purification and Some Properties of the Enzyme Expressed inEscherichia coli

“…The N-carbamyl-L-amino acid amidohydrolase of strain NS671 required Co 2 +, Mn 2 +, or Ni 2 + for its activity, and the requirement was similar to a thermostable enzyme of Bacillus stearothermophilus NS ll22A 7) and fJ-ureidopropionase of P. putida IFOI2996.…”

Purification and Characterization of an ATP‐dependent Amidohydrolase, N‐methylhydantoin Amidohydrolase, from Pseudomonas putida 77

“…The stereospecificity for 5-methylhydantoin is also different; the present enzyme hydrolyzed only the L-isomer, whereas dihydropyrimidinases hydrolyze the D-iSOmer but not the L-isomer [Il,161. Recently, L-isomer-specific hydantoin amidohydrolase and non-stereospecific hydantoin amidohydrolase were found in Bacillus brevis AJ-12299 [24] and Pseudomonus sp. NS671 [25], respectively. These enzymes also required ATP, Mg", and K ' for amide substrate hydrolysis, but detailed characterization of these enzyme proteins has still not been performed.…”

“…NMethylhydantoin amidohydrolase resembles dihydropyrimidinase, which is widely distributed from microorganisms to mammals [lo-161, in that both enzymes hydrolyze hydantoin compounds. In contrast to N-methylhydantoin amidohydrolase, however, dihydropyrimidinase does not require ATP for the hydrolysis of its substrates.Enzymes that catalyze this type of ATP-dependent amidohydrolysis reaction include 5-oxoprolinase [17-211, urea amidolyase [22, 231, L-isomer-specific hydantoinase [24] and non-stereospecific hydantoinase [25], in addition to N-methylhydantoin amidohydrolase. Through extensive studies by Meister and coworkers, it has been shown that rat kidney 5-oxoprolinase is involved in the metabolism of glutathione [26], and that phosphorylation of the substrate is involved in the reaction mechanism 127-291.…”

“…Enzymes that catalyze this type of ATP-dependent amidohydrolysis reaction include 5-oxoprolinase [17-211, urea amidolyase [22, 231, L-isomer-specific hydantoinase [24] and non-stereospecific hydantoinase [25], in addition to N-methylhydantoin amidohydrolase. Through extensive studies by Meister and coworkers, it has been shown that rat kidney 5-oxoprolinase is involved in the metabolism of glutathione [26], and that phosphorylation of the substrate is involved in the reaction mechanism 127-291.…”

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

“…Such inclusion body formation has also been observed during heterologous expression studies on the hyuH and hyuC genes from A. aurescens DSM 3747. 23) From the industrial viewpoint, it would be useful to be able to use not only bacterial strains with native hyu genes, 24) but also recombinant strains that expressing hyu genes. 23,25) In order to utilize recombinant strains for industrial purposes, it will be necessary to establish a strain that expresses hyu genes in optimal proportions.…”