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Applied Microbiology and Biotechnology

, Volume 62, Issue 1, pp 53–60 | Cite as

A novel enzyme, d-3-hydroxyaspartate aldolase from Paracoccus denitrificans IFO 13301: purification, characterization, and gene cloning

  • J. Q. Liu
  • T. Dairi
  • N. ItohEmail author
  • M. Kataoka
  • S. Shimizu
Original Paper

Abstract

A novel enzyme, d-3-hydroxyaspartate aldolase (d-HAA), catalyzing the conversion of d-3-hydroxyaspartate to glyoxylate plus glycine, was purified to homogeneity from Paracoccus denitrificans IFO 13301. d-HAA is strictly d-specific as to the α-position, whereas the enzyme does not distinguish between threo and erythro forms at the β-position. In addition to d-3-hydroxyaspartate, the enzyme also acts on d-threonine, d-3-3,4-dihydroxyphenylserine, d-3-3,4-methylenedioxyphenylserine, and d-3-phenylserine. The d-HAA gene was cloned and sequenced. The gene contains an open reading frame consisting of 1,161 nucleotides corresponding to 387 amino acid residues. The predicted amino acid sequence displayed 35% and 22% identity with that of the d-threonine aldolase of Arthrobacer sp. DK-38 and Alcaligenes xylosoxidan IFO 12669, respectively. This is the first paper reporting both a purified enzyme with d-3-hydroxyaspartate aldolase activity and also its gene cloning.

Keywords

Glyoxylate Aldolase Paracoccus Denitrificans Aldolase Activity Threonine Aldolase 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410CrossRefPubMedGoogle Scholar
  2. Balcar VJ, Jonston GA, Twitchin B (1977) Stereospecificity of the inhibition of l-glutamate and l-aspartate high affinity uptake in rat brain slices by threo-3-hydroxyaspartate. J Neurochem 28:1145–1146PubMedGoogle Scholar
  3. Gibbs RG, Morris JG (1964) Assay and properties of erythro-β-hydroxyasparte aldolase from Micrococcus denitrificans. Biochim Biochem Acta 85:501–503CrossRefGoogle Scholar
  4. Gibbs RG, Morris JG (1965) Purification and properties of erythro-β-hydroxyasparte dehydratase from Micrococcus denitrificans. Biochem J 97:547–554Google Scholar
  5. Higgins DG, Thompson JD, Gibson TJ (1996) Using CLUSTAL for multiple sequence alignments. Methods Enzymol 66:383–402Google Scholar
  6. Ishiyama T, Furuta T, Takai M, Okimoto Y (1975) l- threo-β-hydroxyaspartate acid as an antibiotic amino acid. J Antibiotics 23:821–823Google Scholar
  7. Kaneko T, Katsura H (1963) The synthesis of four optical isomers of β-hydroxyaspartatic acid. Bull Chem Soc Jpn 36:899–930Google Scholar
  8. Kornberg HL, Morris JG (1965) The utilization of glycollate by Micrococcus denitrificans: the β-hydroxyaspartate pathway. Biochem J 95:577–586Google Scholar
  9. Lebrun B, Sakitani M, Shimamoto K, Yasuda-Kamatani Y, Nakajima T (1997) New β-hydroxyaspartate derivatives are competitive blockers for the bovine glutamate/aspartate transporter. J Biol Chem 272:20336–20339CrossRefGoogle Scholar
  10. Liu JQ, Dairi T, Itoh N, Kataoka M, Shimizu S, Yamada H (1998) Novel metal-activated pyridoxal enzyme with a unique primary structure, low-specificity d-threonine aldolase from Arthrobacter sp. strain DK-38. J Biol Chem 273:16678–16685CrossRefPubMedGoogle Scholar
  11. Liu JQ, Dairi T, Itoh N, Kataoka M, Shimizu S, Yamada H (2000a) Diversity of microbial threonine aldolasees and their application. J Mol Cat B Enzymatic 10:107–115Google Scholar
  12. Liu JQ, Otani M, Dairi T, Itoh N, Kataoka M, Shimizu S, Yamada H (2000b) Gene cloning and overproduction of low-specificity d-threonine aldolase from Alcaligenes xylosoxidans and its application for production of a key intermediate for parkinsonism drug. Appl Microbiol Biotechnol 54:44–51CrossRefPubMedGoogle Scholar
  13. Ohashi N, Nagata S, Ishizumi K, Maeshima K (1984) European Patent 83,300,059Google Scholar
  14. Pearson WR, Lipman DJ (1988) Improved tools for biological sequence comparison. Proc Natl Acad Sci USA 85:2444–2448PubMedGoogle Scholar
  15. Saito H, Miura K (1963) Preparation of transforming deoxyribonucleic acid by phenol treatment. Biochim Biophys Acta 72:619–629Google Scholar
  16. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.Google Scholar
  17. Shimamoto K, Shigeri Y, Yasuda-Kamatani Y, Lebrun B, Yumoto N, Nakajima T (2000) Syntheses of optically pure β-hydroxyaspartate derivatives as glutamate transporter blockers. Bioorg Med Chem Lett 10:2407–2410CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • J. Q. Liu
    • 1
    • 3
  • T. Dairi
    • 1
  • N. Itoh
    • 1
    Email author
  • M. Kataoka
    • 2
  • S. Shimizu
    • 2
  1. 1.Biotechnology Research CenterToyama Prefectural UniversityKosugiJapan
  2. 2.Division of Applied Life Sciences, Graduate School of AgricultureKyoto UniversityKyotoJapan
  3. 3.Analytical and Microbiology Department, Kobe Technical CenterProcter and Gamble Far EastKobeJapan

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