Abstract
We studied the biosynthesis of isocitric acid from rapeseed (canola) oil by the yeast Yarrowia lipolytica and its regulation. We determined a fundamental possibility for directed biosynthesis of isocitric acid by Y. lipolytica yeast, with only minimal amounts of citric acid byproduct, when grown on a medium containing canola oil. Wild type strains of Y. lipolytica were mutagenized by UV irradiation and treatment with N-methyl-N′-nitro-N-nitrosoguanidine (NG). Subsequent selection on media with acetate and isocitrate resulted in isolation of a mutant Y. lipolytica UV/NG that synthesized isocitrate and citrate at a ratio of 2.7 : 1. In the parent strain, this ratio is 1 : 1. Inhibition of isocitrate lyase, a key enzyme in the metabolism of isocitric acid, by the addition of itaconic acid resulted in increased synthesis of isocitrate, with a ratio of isocitrate to citrate reaching 6 : 1. The cultivation of the Y. lipolytica UV/NG mutant in a pilot industrial fermenter in the presence of itaconic acid resulted in the production of 88.7 g/L of isocitric acid with a yield of 90%.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Finogenova, T.V., Morgunov, I.G., Kamzolova, S.V., and Chernyavskaya, O.G., Prikl. Biokhim. Mikrobiol., 2005, vol. 41, no. 5, pp. 478–486.
Stottmeister, U., Aurich, A., Wilde, H., Andersch, J., Schmidt, S., and Sicker, D., J. Ind. Microbiol. Biotechnol., 2005, vol. 32, nos. 11–12, pp. 651–664.
Sauer, M., Porro, D., Mattanovich, D., and Branduardi, P., Trends Biotechnol., 2008, vol. 26, no. 2, pp. 100–108.
Aurich, A., Specht, R., Muller, R.A., Stottmeister, U., Yovkova, V., Otto, C., Holz, M., Barth, G., Heretsch, P., Thomas, F.A., Sicker, D., and Giannis, A., Reprogramming Microbial Metabolic Pathways Subcellular Biochemistry, 2012, vol. 64, pp. 391–423.
Groenewald, M., Boekhout, T., Neuveglise, C., Gaillardin, C., van Dijck, P.W., and Wyss, M., Crit. Rev. Microbiol., 2014, vol. 40, no. 3, pp. 187–206.
Rywinska, A., Juszczyk, P., Wojtatowicz, M., Robak, M., Lazar, Z., Tomaszewska, L., and Rymowicz, W., Biomass Bioenergy, 2013, vol. 48, no. 1, pp. 148–166.
Vickery, H.B., J. Biol. Chem., 1962, vol. 237, no. 6, pp. 1739–1741.
Finogenova, T.V., Illarionova, V.I., and Lozinov, A.B., Mikrobiologiya, 1973, vol. 42, no. 4, pp. 790–794.
Lozinov, A.B., Finogenova, T.V., Glazunova, L.M., and Illarionova, V.I., Mikrobiologiya, 1974, vol. 43, no. 5, pp. 786–790.
Finogenova, T.V., Shishkanova, N.V., Fausek, E.A., and Eremina, S.S., Appl. Microbiol. Biotechnol., 1991, vol. 36, no. 2, pp. 231–235.
Kamzolova, S.V., Morgunov, I.G., Aurich, A., Perevoznikova, O.A., Shishkanova, N.V., Stottmeister, U., and Finogenova, T.V., Food Technol. Biotechnol., 2005, vol. 43, no. 2, pp. 113–122.
Kamzolova, S.V., Finogenova, T.V., Lunina, Yu.N., Perevoznikova, O.A., Minachova, L.N., and Morgunov, I.G., Microbiology (Moscow), 2007, vol. 76, no. 1, pp. 20–24.
Forster, A., Jacobs, K., Juretzek, T., Mauersberger, S., and Barth, G., Appl. Microbiol. Biotechnol., 2007, vol. 77, no. 4, pp. 861–869.
Forster, A., Aurich, A., Mauersberger, S., and Barth, G., Appl. Microbiol. Biotechnol., 2007, vol. 75, no. 6, pp. 1409–1417.
Heretsch, P., Thomas, F., Aurich, A., Krautscheid, H., Sicker, D., and Giannis, A., Angew. Chem., Int. Ed. Engl., 2008, vol. 47, no. 10, pp. 1958–1960.
Kamzolova, S.V., Finogenova, T.V., and Morgunov, I.G., Food Technol. Biotechnol., 2008, vol. 46, no. 1, pp. 51–59.
Holz, M., Forster, A., Mauersberger, S., and Barth, G., Appl. Microbiol. Biotechnol., 2009, vol. 81, no. 6, pp. 1087–1096.
Kamzolova, S.V., Lunina, J.N., and Morgunov, I.G., J. Am. Oil Chem. Soc., 1965–1976, vol. 88, no. 12.
Kamzolova, S.V., Dedyukhina, E.G., Samoilenko, V.A., Lunina, J.N., Puntus, I.F., Allayarov, R.K., Chiglintseva, M.N., Mironov, A.A., and Morgunov, I.G., Appl. Microbiol. Biotechnol., 2013, vol. 97, no. 20, pp. 9133–9144.
Burkholder, P., McVeigh, J., and Moyer, D., J. Bacteriol., 1944, vol. 48, no. 4, pp. 385–391.
Velot, Ch., Lebreton, S., Morgunov, I., Usher, K., and Srere, P., Biochemistry, 1999, vol. 38, no. 49, pp. 16195–16204.
Finogenova, T.V., Puntus, I.F., Kamzolova, S.V., Lunina, Yu.N., Monastyrskaya, S.E., Morgunov, I.G., and Boronin, A.M., Appl. Biochem. Microbiol., 2008, vol. 44, no. 2, pp. 197–202.
Berg, I.A., Filatova, L.V., and Ivanovsky, R.N., FEMS Microbiol. Letts., 2002, vol. 216, no. 1, pp. 49–54.
Kratky, M. and Vinsova, J., Curr. Med. Chem., 2012, vol. 19, no. 36, pp. 6126–6137.
Karklin’, R.Ya., Peltsmane, I.Zh., Raminya, L.O., and Korde, G.V., Metabolizm alkanov i sverkhsintez produktov mikroorganizmami (Metabolism of Alkanes and Overproduction of Products by Microorganisms), Finogenova, T.V. and Shar’sheva, A.A, Eds., Pushchino: Pushch. Nauch. Tsentr, 1991.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © S.V. Kamzolova, Y.N. Lunina, R.K. Allayarov, I.F. Puntus, I.A. Laptev, V.A. Samoilenko, I.G. Morgunov, 2015, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2015, Vol. 51, No. 2, pp. 251–257.
Rights and permissions
About this article
Cite this article
Kamzolova, S.V., Lunina, Y.N., Allayarov, R.K. et al. Biosynthesis of isocitric acid by the yeast Yarrowia lipolytica and its regulation. Appl Biochem Microbiol 51, 249–254 (2015). https://doi.org/10.1134/S0003683815020076
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0003683815020076