Amino Acids

, Volume 47, Issue 5, pp 975–985 | Cite as

Biosynthesis of d-aspartate in mammals: the rat and human homologs of mouse aspartate racemase are not responsible for the biosynthesis of d-aspartate

  • Satsuki Matsuda
  • Masumi Katane
  • Kazuhiro Maeda
  • Yuusuke Kaneko
  • Yasuaki Saitoh
  • Tetsuya Miyamoto
  • Masae Sekine
  • Hiroshi HommaEmail author
Original Article


d-Aspartate (d-Asp) has important physiological functions, and recent studies have shown that substantial amounts of free d-Asp are present in a wide variety of mammalian tissues and cells. Biosynthesis of d-Asp has been observed in several cultured rat cell lines, and a murine gene (glutamate-oxaloacetate transaminase 1-like 1, Got1l1) that encodes Asp racemase, a synthetic enzyme that produces d-Asp from l-Asp, was proposed recently. The product of this gene is homologous to mammalian glutamate-oxaloacetate transaminase (GOT). Here, we tested the hypothesis that rat and human homologs of mouse GOT1L1 are involved in Asp synthesis. The following two approaches were applied, since the numbers of attempts were unsuccessful to prepare soluble GOT1L1 recombinant proteins. First, the relationship between the d-Asp content and the expression levels of the mRNAs encoding GOT1L1 and d-Asp oxidase, a primary degradative enzyme of d-Asp, was examined in several rat and human cell lines. Second, the effect of knockdown of the Got1l1 gene on d-Asp biosynthesis during culture of the cells was determined. The results presented here suggest that the rat and human homologs of mouse GOT1L1 are not involved in d-Asp biosynthesis. Therefore, d-Asp biosynthetic pathway in mammals is still an urgent issue to be resolved.


Aspartate racemase d-Aspartate d-Amino acid Amino acid racemase Glutamate-oxaloacetate transaminase 



d-Amino acid oxidase


d-Aspartate oxidase


Glyceraldehyde-3-phosphate dehydrogenase


Glutamate-oxaloacetate transaminase


Glutamate-oxaloacetate transaminase 1-like 1


High-performance liquid chromatography








Pyridoxal phosphate



This work was supported by a Grant-in-Aid for Scientific Research (24590090) from the Japan Society for the Promotion of Science and a Kitasato University Research Grant for Young Researchers (to M.K.).

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Arriza JL, Eliasof S, Kavanaugh MP, Amara SG (1997) Excitatory amino acid transporter 5, a retinal glutamate transporter coupled to a chloride conductance. Proc Natl Acad Sci USA 94:4155–4160CrossRefPubMedCentralPubMedGoogle Scholar
  2. D’Aniello G, Ronsini S, Guida F, Spinelli P, D’Aniello A (2005) Occurrence of d-aspartic acid in human seminal plasma and spermatozoa: possible role in reproduction. Fertil Steril 84:1444–1449CrossRefPubMedGoogle Scholar
  3. D’Aniello G, Grieco N, Di Filippo MA, Cappiello F, Topo E, D’Aniello E, Ronsini S (2007) Reproductive implication of d-aspartic acid in human pre-ovulatory follicular fluid. Hum Reprod 22:3178–3183CrossRefPubMedGoogle Scholar
  4. De Miranda J, Santoro A, Engelender S, Wolosker H (2000) Human serine racemase: molecular cloning, genomic organization and functional analysis. Gene 256:183–188CrossRefPubMedGoogle Scholar
  5. Di Fiore MM, Santillo A, Baccari GC (2014) Current knowledge of d-aspartate in glandular tissues. Amino Acids 46:1805–1818CrossRefPubMedGoogle Scholar
  6. Errico F, Napolitano F, Nisticò R, Usiello A (2012) New insights on the role of free d-aspartate in the mammalian brain. Amino Acids 43:1861–1871CrossRefPubMedGoogle Scholar
  7. Errico F, Napolitano F, Squillace M, Vitucci D, Blasi G, de Bartolomeis A, Bertolino A, D’Aniello A, Usiello A (2013) Decreased levels of d-aspartate and NMDA in the prefrontal cortex and striatum of patients with schizophrenia. J Psychiatr Res 47:1432–1437CrossRefPubMedGoogle Scholar
  8. Fagg GE, Matus A (1984) Selective association of N-methyl aspartate and quisqualate types of L-glutamate receptor with brain postsynaptic densities. Proc Natl Acad Sci USA 81:6876–6880Google Scholar
  9. Hashimoto A, Nishikawa T, Oka T, Takahashi K, Hayashi T (1992) Determination of free amino acid enantiomers in rat brain and serum by high-performance liquid chromatography after derivatization with N-tert-buthloxycarbonyl-l-cysteine and o-phthalaldehyde. J Choromatogr 582:41–48CrossRefGoogle Scholar
  10. Hoffman HE, Jirásková J, Ingr M, Zvelebil M, Konvalinka J (2009) Recombinant human serine racemase: enzymologic characterization and comparison with its mouse ortholog. Protein Expr Purif 63:62–67CrossRefPubMedGoogle Scholar
  11. Kanai Y, Hediger MA (1992) Primary structure and functional characterization of a high-affinity glutamate transporter. Nature 360:467–471CrossRefPubMedGoogle Scholar
  12. Katane M, Homma H (2010) d-Aspartate oxidase: the sole catabolic enzyme acting on free d-aspartate in mammals. Chem Biodivers 7:1435–1449CrossRefPubMedGoogle Scholar
  13. Katane M, Homma H (2011) d-Aspartate—an important bioactive substance in mammals: a review from an analytical and biological point of view. J Chromatogr B 879:3108–3121CrossRefGoogle Scholar
  14. Katane M, Saitoh Y, Seida Y, Sekine M, Furuchi T, Homma H (2010) Comparative characterization of three d-aspartate oxidases and one d-amino acid oxidase from Caenorhabditis elegans. Chem Biodivers 7:1424–1434CrossRefPubMedGoogle Scholar
  15. Katane M, Osaka N, Matsuda S, Maeda K, Kawata T, Saitoh Y, Sekine M, Furuchi T, Doi I, Hirono S, Homma H (2013) Identification of novel d-amino acid oxidase inhibitors by in silico screening and their functional characterization in vitro. J Med Chem 56:1894–1907CrossRefPubMedGoogle Scholar
  16. Kim PM, Duan X, Huang AS, Liu CY, Ming G-L, Song H, Snyder SH (2010) Aspartate racemase, generating neuronal d-aspartate, regulates adult neurogenesis. Proc Natl Acad Sci USA 107:3175–3179CrossRefPubMedCentralPubMedGoogle Scholar
  17. Konno R (2003) Rat cerebral serine racemase: amino acid deletion and truncation at carboxy terminus. Neurosci Lett 349:111–114CrossRefPubMedGoogle Scholar
  18. Long Z, Homma H, Lee J-A, Fukushima T, Santa T, Iwatsubo T, Yamada R, Imai K (1998) Biosynthesis of d-aspartate in mammalian cells. FEBS Lett 434:231–235CrossRefPubMedGoogle Scholar
  19. Long Z, Lee J-A, Okamoto T, Nimura N, Imai K, Homma H (2000) d-Aspartate in a prolactin-secreting clonal strain of rat pituitary tumor cells (GH3). Biochem Biophys Res Commun 276:1143–1147CrossRefPubMedGoogle Scholar
  20. Long Z, Sekine M, Adachi M, Furuchi T, Imai K, Nimura N, Homma H (2002) Cell density inversely regulates d- and l-aspartate levels in rat pheochromocytoma MTP1 cells. Arch Biochem Biophys 404:92–97CrossRefPubMedGoogle Scholar
  21. Nimura N, Kinoshita T (1986) o-Phthalaldehyde-N-acetyl-l-cysteine as a chiral derivatization reagent for liquid chromatographic optical resolution of amino acid enantiomers and its application to conventional amino acid analysis. J Choromatogr 352:169–177CrossRefGoogle Scholar
  22. Nishikawa T (2011) Analysis of free d-serine in mammals and its biological relevance. J Chromatogr B 879:3169–3183CrossRefGoogle Scholar
  23. Ohide H, Miyoshi Y, Maruyama R, Hamase K, Konno R (2011) d-Amino acid metabolism in mammals: biosynthesis, degradation and analytical aspects of the metabolic study. J Chromatogr B 879:3162–3168CrossRefGoogle Scholar
  24. Olverman HJ, Jones AW, Mewett KN, Watkins JC (1988) Structure/activity relations of N-methyl-d-aspartate receptor ligands as studied by their inhibition of [3H]D-2-amino-5-phosphonopentanoic acid binding in rat brain membranes. Neuroscience 26:17–31Google Scholar
  25. Ota N, Shi T, Sweedler JV (2012) d-Aspartate acts as a signaling molecule in nervous and neuroendocrine systems. Amino Acids 43:1873–1886CrossRefPubMedCentralPubMedGoogle Scholar
  26. Pine G, Danbolt NC, Bjøås M, Zheng Y, Bendaham A, Eide L, Koepsell H, Storm-Mathisen J, Seeberg E, Kanner BI (1992) Cloning and expression of a rat brain l-glutamate transporter. Nature 360:464–467CrossRefGoogle Scholar
  27. Shibata K, Watanabe T, Yoshikawa H, Abe K, Takahashi S, Kera Y, Yamada R (2003) Purification and characterization of aspartate racemase from the bivalve mollusk Scapharca broughtonii. Comp Biochem Physiol B Biochem Mol Biol 134:307–314CrossRefPubMedGoogle Scholar
  28. Stříšovský K, Jirásková J, Mikulová A, Rulíšek L, Konvalinka J (2005) Dual substrate and reaction specificity in mouse serine racemase: identification of high-affinity decarboxylate substrate and inhibitor and analysis of the β-eliminase activity. Biochemistry 44:13091–13100CrossRefPubMedGoogle Scholar
  29. Wang L, Ota N, Romanova EV, Sweedler JV (2011) A novel pyridoxal 5′-phosphate-dependent amino acid racemase in the Aplysia californica central nervous system. J Biol Chem 286:13765–13774CrossRefPubMedCentralPubMedGoogle Scholar
  30. Wolosker H (2007) NMDA receptor regulation by d-serine: new findings and perspectives. Mol Neurobiol 36:152–164CrossRefPubMedGoogle Scholar
  31. Wolosker H, Sheth KN, Takahashi M, Mothet J-P, Brady RO Jr, Ferris CD, Snyder SH (1999a) Purification of serine racemase: biosynthesis of the neuromodulator d-serine. Proc Natl Acad Sci USA 96:721–725CrossRefPubMedCentralPubMedGoogle Scholar
  32. Wolosker H, Blackshaw S, Snyder SH (1999b) Serine racemase: a glial enzyme synthesizing d-serine to regulate glutamate-N-methyl-d-aspartate neurotransmission. Proc Natl Acad Sci USA 96:13409–13414CrossRefPubMedCentralPubMedGoogle Scholar
  33. Wolosker H, D’Aniello A, Snyder SH (2000) d-Aspartate disposition in neuronal and endocrine tissues: ontogeny, biosynthesis and release. Neuroscience 100:183–189CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Satsuki Matsuda
    • 1
  • Masumi Katane
    • 1
  • Kazuhiro Maeda
    • 1
  • Yuusuke Kaneko
    • 1
  • Yasuaki Saitoh
    • 1
  • Tetsuya Miyamoto
    • 1
  • Masae Sekine
    • 1
  • Hiroshi Homma
    • 1
    Email author
  1. 1.Laboratory of Biomolecular Science, Graduate School of Pharmaceutical SciencesKitasato UniversityTokyoJapan

Personalised recommendations