Higher plant cytosolic er-based glycerol-3-phosphate...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/54 (2006.01) A01H 1/00 (2006.01) A01H 1/04 (2006.01) A01H 5/00 (2006.01) A01H 5/10 (2006.01) C07H 21/00 (2006.01) C12N 5/04 (2006.01) C12N 5/10 (2006.01) C12N 9/10 (2006.01) C12N 15/82 (2006.01) C12P 7/64 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2461050

Glycerol-3-phosphate acyltransferase is the initial enzyme of the glycerolipid biosynthetic pathway. Biochemical analyses indicated that the reaction mediated by glycerol-3-phosphate acyltransferase represents a potential rate- limiting step for the synthesis of phospholipids and storage neutralipid, triacylglycerol. The present invention relates to the cloning of genes encoding extraplastidic membrane-bound glycerol-3-phosphate acyltransferases. Heterologous expression of the genes, GPAT1, GPAT2, and GPAT3 in a yeast glycerol-3-phosphate acyltransferase mutant demonstrated that the encoded products could efficiently utilize glycerol-3-phosphate to mediate sn-1 stereo- specific fatty acid acylation. The invention encompasses the glycerol-3- phosphate acyltransferase peptides disclosed and fragments and homologues thereof, the corresponding gene sequences and fragments and homologues thereof, as well as the use of the peptide and gene sequences of the present invention for use in generating recombinant proteins, and transgenic plants with altered lipid metabolism. In this way, the present invention also encompasses the use of such recombinant peptides and transgenic plants for the production of lipid products for use, for example, in pharmaceutical and nutritional applications.

la glycérol-3-phosphate acyltransférase est l'enzyme de départ de la voie biosynthétique des glycérolipides. D'après des analyses biochimiques, il semble que la réaction induite par la glycérol-3-phosphate acyltransférase représente une opération de limitation de taux potentielle pour la synthèse de phospholipides et du neutralipide de stockage, le triacylglycérol. La présente invention concerne le clonage de gènes codant pour des glycérol-3-phosphate acyltransférases liés à la membrane extraplastidique. L'expression hétérologue des gènes, <i>GPAT1, GPAT2</i>, et <i>GPAT3</i> dans un mutant de glycérol-3-phosphate acyltransférase de levure a démontré que les produits codés pouvaient utiliser efficacement le glycerol-3-phosphate pour induire une acylation stéréospécifique des acides gras en position <i>sn-1</i>. Cette invention englobe les peptides de glycérol-3-phosphate acyltransférase de l'invention ainsi que des fragments et des homologues de ces peptides, les séquences de gène correspondantes et des fragments et des séquences de ces séquences, ainsi que l'utilisation des séquences de peptides et de gènes de ladite invention pour la réaction de protéines de recombinaison, et des plantes transgéniques au métabolisme lipidique modifié. Ainsi, cette invention porte également sur l'emploi de tels peptides de recombinaison et de plantes transgéniques pour la production de lipides destinés, par exemples, à des applications pharmaceutiques et nutritionnelles.

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