Abstract
In an attempt to clarify the mechanism of lipid accumulation inMortierella ramanniana var.angulispora, diacylglycerol acyltransferase (DGAT) in the membrane fraction from this fungus was characterized. The enzyme had an optimum pH of 7.0–7.5, and enzyme activity was blocked by SH-reagents. Metal ions were not essential for maintaining DGAT activity.n-Octyl-β-d-glucoside, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate and Tween 80 were found to preserve activity, while Triton X-100 and sucrose monolaurate inhibited it. As the inhibition of DGAT activity by Triton X-100 was overcome by the addition of diacylglycerol (DG), the dependency of DGAT activity on exogenous DG was determined in the presence of 0.1% Triton X-100. DGAT activity in the membrane fraction was traced in fungi cultured for different time periods or in media at different carbon to nitrogen (C/N) ratios. Although the increase in total lipid content with culture time was accompanied by an increase in DGAT activity, total lipid changes related to changes in C/N ratio did not correlate with DGAT activity. Factors other than DGAT activity in the membrane fraction would appear to be involved in the regulation of total lipid content in this fungus.
Similar content being viewed by others
Abbreviations
- CHAPS:
-
3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate
- C/N ratio:
-
carbon to nitrogen ratio
- DG:
-
diacylglycerol
- DGAT:
-
diacylglycerol acyltransferase
- EDTA:
-
ethylenediaminetetraacetic acid
- TG:
-
triacylglycerol
- TLC:
-
thin-layer chromatography
References
Tijburg, L.B.M., Geelen, M.J.H., and van Golde, L.M.G. (1989)Biochim. Biophys. Acta 1004, 1–19.
Martin, B.A., and Wilson, R.F. (1983)Lipids 18, 1–6.
Ichihara, K., and Noda, M. (1982)Phytochemistry 21, 1895–1901.
Christiansen, S. (1979)Biochim. Biophys. Acta 574, 448–460.
Goldberg, D.M., Roomi, M.W., Yu, A., and Roncari, D.A.K. (1981)Biochem. J. 196, 337–346.
Haagsman, H.P., and van Golde, L.M.G. (1981)Arch. Biochem. Biophys. 208, 395–402.
Mayorek, N., Grinstein, I., and Bar-Tana, J. (1989)Eur. J. Biochem. 182, 395–400.
Brindley, D.N. (1984)Prog. Lipid Res. 23, 115–153.
Ichihara, K., Murota, N., and Fujii, S. (1990)Biochim. Biophys. Acta 1043, 227–234.
Hosaka, K., and Yamashita, S. (1984)Biochim. Biophys. Acta 796, 110–117.
Yamashita, S., Hosaka, K., Miki, Y., and Numa, S. (1981)Methods Enzymol. 71, 528–536.
Kwanyuen, P., Wilson, R.F., and Burton, J.W. (1988) inBiotechnology for the Fats and Oils Industry (Applewhite, T.E., ed.) pp. 294–297, American Oil Chemists’ Society, Champaign.
Bafor, M., Stobart, A.K., and Stymne, S. (1990)J. Am. Oil Chem. Soc. 67, 217–225.
Suzuki, O. (1988) inBiotechnology for the Fats and Oils Industry (Applewhite, T.E., ed.) pp. 110–116, American Oil Chemists’ Society, Champaign.
Kamisaka, Y., Kikutsugi, H., Yokochi, T., Nakahara, T., and Suzuki, O. (1988)J. Jpn. Oil Chem. Soc. (Yukagaku) 37, 344–348.
Yokochi, T., and Suzuki, O. (1989)J. Jpn. Oil Chem. Soc. (Yukagaku) 38, 1007–1015.
Kamisaka, Y., Yokochi, T., Nakahara, T., and Suzuki, O. (1990)Lipids 25, 54–60.
Kamisaka, Y., Yokochi, T., Nakahara, T., and Suzuki, O. (1990)Lipids 25, 787–792.
Bligh, E.G., and Dyer, W.J. (1959)Can. J. Biochem. Physiol. 37, 911–917.
Jigami, Y., Suzuki, O., and Nakasato, S. (1979)Lipids 14, 937–942.
Bradford, M. (1976)Anal. Biochem. 72, 248–254.
Ichihara, K., Takahashi, T., and Fujii, S. (1988)Biochim. Biophys. Acta 958, 125–129.
Cao, Y.-Z., and Huang, A.H.C. (1986)Plant Physiol. 82, 813–820.
Bell, R.M., and Coleman, R.A. (1980)Annu. Rev. Biochem. 49, 459–487.
Coleman, R., and Bell, R.M. (1976)J. Biol. Chem. 251, 4537–4543.
Hosaka, K., Schiele, U., and Numa, S. (1977)Eur. J. Biochem. 76, 113–118.
Kwanyuen, P., and Wilson, R.F. (1986)Biochim. Biophys. Acta 877, 238–245.
Weiss, S.B., Kennedy, E.P., and Kiyasu, J.Y. (1960)J. Biol. Chem. 235, 40–44.
Ratledge, C. (1989) inMicrobial Lipids (Ratledge, C., and Wilkinson, S.G., eds.) Vol. 2, pp. 567–668, Academic Press, London.
Suzuki, O., Yukochi, T., and Yamashina, T. (1982)Yukagaku 31, 921–931.
Hasson, L., and Dostalek, M. (1988)Appl. Microbiol. Biotechnol. 28, 240–246.
Jigami, Y., Suzuki, O., and Nakasato, S. (1979)Yukagaku 28, 867–869.
Author information
Authors and Affiliations
About this article
Cite this article
Kamisaka, Y., Yokochi, T., Nakahara, T. et al. Characterization of the diacylglycerol acyltransferase activity in the membrane fraction from a fungus. Lipids 28, 583–587 (1993). https://doi.org/10.1007/BF02536050
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02536050