3178
T. OURA and S. KAJIWARA
A
B
80
100
80
60
40
20
0
70
60
50
40
30
20
10
0
PC
sn-1 sn-2
PC PC
PE
sn-1 sn-2
PE PE
PC
sn-1 sn-2
PC PC
PE
sn-1
PE
sn-2
PE
Fraction
Fraction
Fig. 4. ꢀ12 (A) and !3 (B) Desaturation at the sn-1 and sn-2 Positions in PC and PE in the S. kluyveri IFO1893 Strain.
Desaturase activities were calculated as (C18:2 + ꢀ-C18:3)/(C18:1 + C18:2 + ꢀ-C18:3) ꢁ 100 for Sk-FAD2, and as ꢀ-C18:3/(C18:2 +
ꢀ-C18:3) ꢁ 100 for Sk-FAD3, using values corresponding to the percentage of total fatty acids.
9) Ferrante, G., and Kates, M., Pathways for desaturation of
oleoyl chains in Candida lypolytica. Can. J. Biochem.
Cell Biol., 61, 1191–1196 (1983).
of the fungi, S. kluyveri, can recognize lipid substrates
in a manner different from ꢀ5 and ꢀ6 fatty acid
desaturases.
10) Sperling, P., Linscheid, M., Stocker, S., Muhlbach, H. P.,
and Heinz, E., In vivo desaturation of cis-delta 9-
monounsaturated to cis-delta 9,12-diunsaturated alkeny-
lether glycerolipids. J. Biol. Chem., 268, 26935–26940
(1993).
11) Los, D. A., and Murata, N., Structure and expression
of fatty acid desaturase. Biochim. Biophys. Acta, 1394,
3–15 (1998).
Acknowledgments
This work was financed by a Grant for Young
Scientists (no. 18880011) from the Japan Society for
the Promotion of Science.
12) Wallis, J. G., and Browse, J., Mutants of Arabidopsis
reveal many roles for membrane lipids. Prog. Lipid Res.,
41, 254–278 (2002).
13) Griffiths, G., Stobart, A. K., and Stymne, S., Delta 6- and
delta 12-desaturase activities and phosphatidic acid
formation in microsomal preparations from the devel-
oping cotyledons of common borage (Borago officina-
lis). Biochem. J., 252, 641–647 (1988).
14) Jackson, F. M., Fraser, T. C., Smith, M. A., Lazarus, C.,
Stobart, A. K., and Griffiths, G., Biosynthesis of C18
polyunsaturated fatty acids in microsomal membrane
preparations from the filamentous fungus Mucor circi-
nelloides. Eur. J. Biochem., 252, 513–519 (1998).
15) Galle-Le Bastard, A. M., Demandre, C., Oursel, A.,
Joseph, M., Mazliak, P., and Kader, J. C., Phosphati-
dylcholine molecular species involved in g-linolenic acid
biosynthesis in microsomes from borage seeds. Physiol.
Plant., 108, 118–124 (2000).
References
1) Kinsella, J. E., Lokesh, B., Broughton, S., and Whelan,
J., Dietary polyunsaturated fatty acids and eicosanoids:
potential effects on the modulation of inflammatory and
immune cells: an overview. Nutrition, 6, 24–44 (1990).
2) Wallis, J. G., Watts, J. L., and Browse, J., Polyunsatu-
rated fatty acid synthesis: what will they think of next?
Trends Biochem. Sci., 27, 467–473 (2002).
3) Tocher, D. R., Leaver, M. J., and Hodgson, P. A., Recent
advances in the biochemistry and molecular biology of
fatty acyl desaturases. Prog. Lipid Res., 37, 73–117
(1998).
4) Sperling, P., Ternes, P., Zank, T. K., and Heinz, E., The
evolution of desaturases. Prostaglandins Leukot. Essent.
Fatty Acids, 68, 73–95 (2003).
5) Schwartzbeck, J. L., Jung, S., Abbott, A. G., Mosley, E.,
Lewis, S., Pries, G. L., and Powell, G. L., Endoplasmic
oleoyl-PC desaturase references the second double bond.
Phytochemistry, 57, 643–652 (2001).
16) Domergue, F., Abbadi, A., Ott, C., Zank, T. K.,
Zahringer, U., and Heinz, E., Acyl carriers used as
¨
6) Sasata, R. J., Reed, D. W., Loewen, M. C., and Covello,
P. S., Domain swapping localizes the structural deter-
minants of regioselectivity in membrane-bound fatty
acid desaturases of Caenorhabditis elegans. J. Biol.
Chem., 279, 39296–39302 (2004).
substrates by the desaturases and elongases involved in
very long-chain polyunsaturated fatty acids biosynthesis
reconstituted in yeast. J. Biol. Chem., 278, 35115–35126
(2003).
17) Oura, T., and Kajiwara, S., Saccharomyces kluyveri
FAD3 encodes an !3 fatty acid desaturase. Micro-
biology, 150, 1983–1990 (2004).
7) Meesapyodsuk, D., Reed, D. W., Savile, C. K., Buist,
P. H., Schafer, U. A., Ambrose, S. J., and Covello, P. S.,
¨
Substrate specificity, regioselectivity and cryptoregio-
chemistry of plant and animal !-3 fatty acid desaturases.
Biochem. Soc. Trans., 28, 632–635 (2000).
18) Watanabe, K., Oura, T., Sakai, H., and Kajiwara, S.,
Yeast ꢀ12 fatty acid desaturase: gene cloning expression
and function. Biosci. Biotechnol. Biochem., 68, 721–727
(2004).
8) Reed, D. W., Schafer, U. A., and Covello, P. S.,
¨
Characterization of the Brassica napus extraplastidial
linoleate desaturase by expression in Saccharomyces
cerevisiae. Plant Physiol., 122, 715–720 (2000).
19) Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D.,
Seidman, J. G., Smith, J. A., and Struhl, K., ‘‘Current
Protocols in Molecular Biology,’’ Green Publishing