Abstract
Two independent mutant strains of methylotrophic yeast Pichia methanolica (mth1 arg1 and mth2 arg4) from the initial line 616 (ade1 ade5) were investigated. The mutant strains possessed defects in genes MTH1 and MTH2 which resulted in the inability to assimilate methanol as a sole carbon source and the increased activity of alcohol oxidase (AO). The function of the AUG2 gene encoding one of the subunits of AO and CTA1, a probable homolog of peroxisomal catalase of Saccharomyces cereviseae, was investigated by analyses of the molecular forms of isoenzymes. It was shown that optimal conditions for the expression of the AUG2 gene on a medium supplemented with 3% of methanol leads to an increasing synthesis of peroxisomal catalase. The mutant mth1 possessed a dominant formation of AO isoform with electrophoretic mobility which is typical for isogenic form 9, the product of the AUG2 gene, and a decreased level of peroxisomal catalase. The restoration of growth of four spontaneous revertants of the mutant mth1 (Rmth1) on the methanol containing medium was accompanied by an increase in activity of AO isogenic form 9 and peroxisomal catalase. The obtained results confirmed the functional continuity of the structural gene AUG2 in mutant mth1. The correlation of activity of peroxisomal catalase and AO isogenic form 1 in different conditions evidenced the existence of common regulatory elements for genes AUG2 and CTA1 in methilotrophic yeast Pichia methanolica.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Sibirny, A.A., Titorenko, V.I., Efremov, B.D., and Tolstorukov, I.I., Yeast, 1987, vol. 3, pp. 233–241.
Raymond, C.K., in Gene Expression Systems, Using Nature of the Art of Expression, Fernandez, J. M. and Hoeffler, J. P., Eds., New York: Academic, 1999, pp. 193–209.
Johnson, M.A., Waterham, H.R., Ksheminska, G.P., Fayura, L.R., Cereghino, J.L., Stasyk, O.V., Veenhuis, M., Kulachkovsky, A.R., Sibirny, A.A., and Cregg, J.M., Genetics, 1999, vol. 151, pp. 1379–1391.
Nakagawa, T., Miyaji, T., Yurimoto, H., Sakai, Y., Kato, N., and Tomizuka, N., Appl. Environ. Microbiol., 2000, vol. 66, pp. 4253–4257.
Nakagawa, T., Yamada, K., Fujimura, S., Ito, T., Miyaji, T., and Tomizuka, N., Microbiology, 2005, vol. 151, pp. 2047–2052.
Sahm, H. and Wagner, F., Eur. J. Biochem., 1973, vol. 36, pp. 250–256.
Veenhuis, M., van Dijen, J.P., and Harder, W., Adv. Microb. Physiol., 1983, vol. 24, pp. 1–82.
Sahm, H., Adv. Biochem. Eng., 1997, vol. 6, pp. 77–81.
Raymond, C.K., Bukowski, T., Holderman, S.D., Ching, A.F., Vanaja, E., and Stamm, M.R., Yeast, 1998, vol. 14, pp. 11–23.
Nakagawa, T., Uchimura, T., and Komagata, K., J. Ferment. Bioeng., 1996, vol. 81, pp. 498–503.
Nakagawa, T., Mukaiyama, H., Yurimoto, H., Sakai, Y., and Kato, N., Yeast, 1999, vol. 15, pp. 1223–1230.
Gruzman, M.B., Titorenko, V.I., Lusta, K.A., and Trotsenko, Yu.A., Biokhimiya, 1996, vol. 61, pp. 2131–2139.
Nakagawa, T., Mizumura, T., Mukaiyama, H., Miyaji, T., Yurimoto, H., Kato, N., Sakai, Y., and Tomizuka, N., Yeast, 2002, vol. 12, pp. 1067–1073.
Roggenkamp, R., Janowicz, Z., Stanikowski, B., and Hollenberg, C.P., Mol. Gen. Genet., 1984, vol. 194, pp. 489–497.
Harder, W. and Veenhuis, M., The Yeast, London: Academic, 1989, vol. 3, pp. 289–316.
Melik-Adamyan, W.R., Barynin, V.V., and Vagin, A.A., J. Mol. Biol., 1986, vol. 188, pp. 63–72.
Cohen, G., Rapatz, W., and Ruis, H., Eur. J. Biochem., 1988, vol. 176, pp. 159–163.
Gasch, A.P., Spellman, P.T., Kao, C.M., Carmel-Harel, O., Eisen, M.B., Storz, G., Botstein, D., and Brown, P.O., Mol. Biol. Cell, 2000, vol. 11, pp. 4241–4257.
Bissinger, P.H., Wieser, R., Hamilton, B., and Ruis, H., Mol. Cell. Biol., 1989, vol. 9, pp. 1309–1315.
Motruk, O.M., Tolstorukov, I.I., and Sibirnyi, A.A., Biotekhnologiya, 1989, vol. 5, no. 6, pp. 692–698.
Tolstorukov, I.I., Dutova, T.A., Benevolenskii, S.V., and Soom, Ya.O., Genetika, 1977, vol. 13, pp. 322–326.
Sibirnyi, A.A., Titorenko, V.I., Benevolenskii, S.V., and Tolstorukov, I.I., Genetika, 1986, no. 12, pp. 584–592.
Shol’ts, K.F. and Ostrovskii, D.N., Metody sovremennoi biokhimii (Methods of Modern Biochemistry), Moscow: Nauka, 1975.
Laemmli, U.K., Nature, 1970, vol. 227, pp. 680–685.
Lledias, F., Rangel, P., and Hansberg, W., J. Biol. Chem., 1998, vol. 273, pp. 10630–10637.
Egli, T., van Dijken, J.P., Veenhuis, M., Harder, W., and Fietcher, A., Arch. Microbiol., 1980, vol. 124, pp. 115–121.
Egli, T., Lindley, N.D., and Quayle, J.R., J. Gen. Microbiol., 1983, vol. 129, p. 1269.
De Coning, W., Glesson, M.A.G., Harder, W., and Dijkhuiren, C., Arch. Microbiol., 1987, vol. 147, pp. 375–382.
Nakagawa, T., Sakai, Y., Mukaiyama, H., Mizumura, T., Miyaji, T., Yurimoto, H., Kato, N., and Tomizuka, N., J. Biosci. Bioeng, 2001, vol. 91, pp. 225–227.
Leonovich, O.A., Pogutse, O.M., Serkova, N.N., Dutova, T.A., Tolstorukov, I.I., and Rabinovich, Ya.M., Biotekhnologiya, 2000, no. 3, pp. 9–15.
Author information
Authors and Affiliations
Corresponding author
Additional information
Original Russian Text © O.A. Leonovich, Yu.A. Kurales, T.A. Dutova, E.P. Isakova, Y.I. Deryabina, Ya.M. Rabinovich, 2009, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2009, Vol. 45, No. 2, pp. 156–162.
Rights and permissions
About this article
Cite this article
Leonovich, O.A., Kurales, Y.A., Dutova, T.A. et al. The regulation of peroxisomal matrix enzymes (alcohol oxidase and catalase) formation by the product of the gene Mth1 in methylotrophic yeast Pichia methanolica . Appl Biochem Microbiol 45, 137–142 (2009). https://doi.org/10.1134/S0003683809020045
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S0003683809020045