Applied Biochemistry and Microbiology

, Volume 53, Issue 2, pp 217–221 | Cite as

Synthesis of L-lactate oxidaze in yeast Yarrowia lipolytica during submerged cultivation

  • E. N. Biryukova
  • A. Yu. Arinbasarova
  • A. G. Medentsev


The biosynthesis of L-lactate oxidase in the Yarrowia lipolytica yeast during submerged cultivation in laboratory bioreactors ANKUM-2M has been studied. It has been shown under optimal conditions of yeast cultivation with L-lactate that 24.5 U/L enzyme accumulated in the medium and the yield was 2.0 U/(L h). An increase in the biosynthesis of L-lactate oxidase to 75 U/L and the yield to 3.2 U/(L h) was achieved in the medium with L-lactate (1%) and glucose (2%). The enzyme was purified 251 times to homogeneity by hydrophobic and ion exchange chromatography state with a yield of 45% and a specific activity of 55.3 U/mg. Techniques of gel filtration and denaturing electrophoresis showed that L-lactate oxidase from Y. lipolytica is a tetramer with a molecular mass of 200–230 kDa. The enzyme showed a strict specificity to L-lactate and did not oxidize fumarate, pyruvate, succinate, ascorbate, dihydroxyacetone, glycolate, D-lactate, D, L-2-hydroxybutyrate and D, L-alanine or D-serine.


L-lactate oxidase yeast Yarrowia lipolytica submerged cultivation 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Duncan, J.D., Wallis, J.O., and Azari, M.R., Biochem. Biophys. Res. Commun., 1989, vol. 164, no. 2, pp. 919–926.CrossRefPubMedGoogle Scholar
  2. 2.
    Takemori, S., Nakazawa, K., Nakai, Y., Suzuki, K., and Katagiri, M.A., J. Biol. Chem., 1968, vol. 243, no. 2, pp. 313–319.PubMedGoogle Scholar
  3. 3.
    Tzanetakis, N. and Litopoulou-Tzanetaki, E., J. Dairy Sci., 1989, vol. 72, pp. 859–863.CrossRefGoogle Scholar
  4. 4.
    Atsusi, T. and Yoshiaki, N., J. Ferment. Bioeng., 1998, vol. 85, no. 5, pp. 507–510.CrossRefGoogle Scholar
  5. 5.
    Sullivan, P.A., Soon, C.Y., Schreurs, W.J., Cutfield, J.F., and Shepherd, M.G., Biochem. J., 1977, vol. 165, no. 2, pp. p. 375–383.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Kupletskaya, M.B., Sukhacheva, M.V., Kurakov, A.V., and Netrusov, A.I., Appl. Biochem. Microbiol., 2007, vol. 43, no. 2, pp. 178–181.CrossRefGoogle Scholar
  7. 7.
    Sztajer, H., Wang, W., Lunsdorf, H., Stocker, A., and Schmid, R.D., Appl. Microbiol. Biotechnol., 1996, vol. 45, no. 5, pp. p. 600–606.CrossRefPubMedGoogle Scholar
  8. 8.
    Biryukova, E.N., Stupar’, Ya.O., Arinbasarova, A.Yu., and Medentsev, A.G., Microbiology (Moscow), 2009, vol. 78, no. 5, pp. 650–652.CrossRefGoogle Scholar
  9. 9.
    Streitenberger, S.A., Lopez-Mas, J.A., Sanchez-Ferrer, A., and Garcia-Carmona, F., Biotechnol. Prog., 2002, vol. 18, no. 3, pp. p. 657–659.CrossRefPubMedGoogle Scholar
  10. 10.
    Esders, T.W., Goodhue, C.T., and Schubert, R.M., US Patent no. 4166763, 1979.Google Scholar
  11. 11.
    Bradford, M.M., A, Anal. Biochem., 1976, vol. 72, pp. 248–254.CrossRefGoogle Scholar
  12. 11a.
    Laemmli, U.K., Nature, 1970, vol. 227, pp. 680–685.CrossRefPubMedGoogle Scholar
  13. 11b.
    Garvie, E.I., Microbiol. Rev., 1980, vol. 44, no. 1, pp. 106–139.PubMedPubMedCentralGoogle Scholar
  14. 12.
    Ghisla, S. and Massey, V., in Chemistry and Biochemistry of Flavoenzymes. II, Muller, F., Ed., Boca Raton, FL: CRC Press, 1991, pp. 243–289.Google Scholar
  15. 13.
    Labeyrie, F., Baudras, A., and Lederer, F., Methods Enzymol., 1978, vol. 53, pp. 238–256.CrossRefPubMedGoogle Scholar

Copyright information

© Pleiades Publishing, Inc. 2017

Authors and Affiliations

  • E. N. Biryukova
    • 1
  • A. Yu. Arinbasarova
    • 1
  • A. G. Medentsev
    • 1
  1. 1.Institute of the Biochemistry and Physiology of MicroorganismsRussian Academy of SciencesPushchino, Moscow oblastRussia

Personalised recommendations