Mechanism of high D-aspartate production in the lactic acid bacterium Latilactobacillus sp. strain WDN19.

D-Aspartate (D-Asp) is a useful compound for a semisynthetic antibiotic and has potentially beneficial effects on humans. Several lactic acid bacteria (LAB) species produce D-Asp as a component of cell wall peptidoglycan. We previously isolated a LAB strain (named strain WDN19) that can extracellularly produce a large amount of D-Asp.

Development of an enzymatic screening method for d-aspartate-producing lactic acid bacteria.

d-Aspartate (d-Asp) is an important intermediate for synthetic penicillin and an endogenous amino acid that plays important roles in the endocrine and nervous systems in animals including humans. Lactic acid bacteria (LABs) have been used as probiotics in humans, and some LAB species produce d-Asp as a component of cell wall peptidoglycan. LAB strains with greater d-Asp production would therefore be valuable for industrial d-Asp production.

d-Aspartate N-methyltransferase catalyzes biosynthesis of N-methyl-d-aspartate (NMDA), a well-known selective agonist of the NMDA receptor, in mice.

N-Methyl-d-aspartate (NMDA), which is a selective agonist for the NMDA receptor, has recently been shown to be present in various biological tissues. In mammals, the activity of d-aspartate N-methyltransferase (DDNMT), which produces NMDA from d-aspartate, has been detected only in homogenates prepared from rat tissues. Moreover, the enzymatic properties of DDNMT have been poorly studied and its molecular entity has not yet been identified.

Liquid chromatography-electrospray ionization-tandem mass spectrometric assay for d-aspartate -methyltransferase activity in ark shells.

A liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method for the separation and quantification of the enantiomers of -methylaspartate and -methylglutamate, after derivatization with -(5-fluoro-2,4-dinitrophenyl)-L-leucinamide was established. The time required for the LC-ESI-MS/MS analysis was within 20 min and the detection limit was approximately 10 fmol per injection, demonstrating that this method can be used for the rapid determination of D-aspartate -methyltransferase activity in the ark shell clam . NMDA: -methyl-D-aspartate; NMLA: -methyl-L-aspartate; NMDG: -methyl-D-glutamate; NMLG: -methyl-L-glutamate; NMA: -methylaspartate; NMG: -methylglutamate; HPLC: high-performance liquid chromatography; SAM: -adenosyl-L-methionine; OPA: -phthalaldehyde; LC-ESI-MS/MS: liquid chromatography-electrospray ionization-tandem mass spectrometry; FDLA: -(5-fluoro-2,4-dinitrophenyl)-L-leucinamide; FDAA: -(5-fluoro-2,4-dinitrophenyl)-L-alaninamide; ESI: electrospray ionization; LC-ESI-MS: liquid chromatography-electrospray ionization-mass spectrometry; MS/MS: tandem mass spectrometry.

Aspartate racemase and D-aspartate in starfish; possible involvement in testicular maturation.

D-Aspartate, aspartate racemase activity, and D-aspartate oxidase activity were detected in tissues from several types of starfish. Aspartate racemase activity in male testes of was significantly elevated in the summer months of the breeding season compared with spring months. We also compared aspartate racemase activity with the gonad index and found that activity in individuals with a gonad index ≥6% was four-fold higher than that of individuals with a gonad index <6%.

Determination of D-aspartate N-methyltransferase activity in the starfish by direct analysis of N-methyl-D-aspartate with high-performance liquid chromatography.

We describe a method for the detection and quantification of D-aspartate N-methyltransferase activity. The enzyme catalyzes the S-adenosyl-L-methionine-dependent N-methylation of D-aspartate to form N-methyl-D-aspartate (NMDA). NMDA is detected directly by high-performance liquid chromatography (HPLC) of their (+)- and/or (-)-1-(9-fluorenyl)ethyl chloroformate fluorescent derivatives.

Nucleotides modulate the activity of aspartate racemase of Scapharca broughtonii.

The activity of D-aspartate racemase purified from Scapharca broughtonii has been found to depend markedly on some nucleotides. Purine nucleoside monophosphates enhanced the enzyme activity, which was, on the contrary, lowered by purine nucleoside triphosphates and not affected by pyrimidine nucleotides. AMP produced the highest increase of seven-fold in the enzyme activity at 6 mM and a half-maximum increase at approximately 3.

Purification and characterization of aspartate racemase from the bivalve mollusk Scapharca broughtonii.

High concentrations of D-aspartate occur in blood shell Scapharca broughtonii (Mollusca) tissues. We purified aspartate racemase from the foot muscle of the bivalve to electrophoretic homogeneity. The molecular mass shown by sodium dodecyl sulfate polyacrylamide gel was 39 kDa, while that shown by gel filtration ranged from 51 to 63 kDa.

N-methyl-D-glutamate and N-methyl-L-glutamate in Scapharca broughtonii (Mollusca) and other invertebrates.

The presence of N-methyl-D-glutamate (NMDG) and N-methyl-L-glutamate (NMLG) has been demonstrated in the tissues of Scapharca broughtonii, which are known to contain N-methyl-D-aspartate (NMDA). To our knowledge, this is the first report on the natural occurrence of NMDG and the occurrence of NMLG in eukaryotes. These compounds were identified according to the following findings; (a) their derivatives with (+)- and (-)-l-(9-fluorenyl)ethyl chloroformate (FLEC) showed identical behaviors with those of authentic NMDG and NMLA, respectively, on high-performance liquid chromatography (HPLC), (b) the HPLC peak of NMDG disappeared when the extract, as well as the authentic compound, was treated with D-aspartate oxidase before derivatization, (c) they behaved identically with authentic compounds on thin-layer chromatography and differently from NMDA.