Calgene Corporation

Monsanto Technology LLC

Kathryn Dennis Lardizabal

Michael W Lassner

James George Metz

Gregory A Thompson

Deborah J Hawkins

Nicholas Wagner

Kristen A Bennett

David Richard Corbin

Ganesh M Kishore

Balasulojini Karunanandaa

Sekhar S Boddupalli

Mylavarapu Venkatramesh

Shaukat H Rangwala

B Ganesh Bhat

Robert J Grebenok

The present invention is directed to polypeptides and nucleic acid sequences related thereto, and methods to purify, obtain, and use such molecules in genetic engineering applications. More specifically, the present invention relates to polypeptides associated with the production of triacylglycerols in plants and fungi.

Diacylglycerol acyltransferase nucleic acid sequences and associated products

The invention provides diacylglycerol acyltransferase (DAGAT) proteins, wherein said proteins are active in the formation of triacylglycerol from fatty acyl and diacylglycerol substrates. In one aspect,DAGAT proteins have been isolated and have molecular weights of between approximately 36 and 37 kDa as measured by SDS-PAGE. The invention also provides novel DAGAT polynucleotide and polypeptide sequences and to methods of producing such polypeptides using recombinant techniques. In addition, methods are provided for using such sequences to alter triacylglycerol levels in plants and to treat diseases associated with altered DAGAT activity or expression.

Diacylglycerol acyl transferase proteins

Provided are recombinant constructs comprising DNA sequences encoding enzymes effective in altering the biosynthesis and accumulation of sterol compounds and tocopherols in transgenic plants. Also provided are methods of using such constructs to produce transgenic plants, seeds of which contain elevated levels of sitostanol and/or sitostanol esters, and &#x3b1;-tocopherol, as well as reduced levels of campesterol and campestanol and their corresponding esters. These seeds also contain the novel sterol brassicastanol. Oil obtained from seeds of such transgenic plants is also provided. This oil can be used to prepare food and pharmaceutical compositions effective in lowering the level of low density lipoprotein cholesterol in blood serum. In addition, novel DNA sequences encoding plant steroid 5&#x3b1;-reductases are also disclosed.

Transgenic plants containing altered levels of sterol compounds and tocopherols

By this invention, nucleic acid sequences encoding for fatty acyl-CoA:fatty alcohol acyltransferase (wax synthase) are provided, wherein said wax synthase is active in the formation of a wax ester from fatty alcohol and fatty acyl-CoA substrates. Of special interest are nucleic acid sequences obtainable from a jojoba embryo wax synthase having an apparent molecular mass of approximately 33 kD. Also considered are amino acid and nucleic acid sequences obtainable from wax synthase proteins and the use of such sequences to provide transgenic host cells capable of producing wax esters.

Fatty acyl-CoA: fatty alcohol acyltransferases

The invention provides diacylglycerol acyltransferase (DAGAT) proteins, wherein said proteins are active in the formation of triacylglycerol from fatty acyl and diacylglycerol substrates. In one aspect,DAGAT proteins have been isolated and have molecular weights of between approximately 36 and 37 kDa as measured by SDS-PAGE. The invention also provides novel DAGAT polynucleotide and polypeptide sequences and to methods of producing such polypeptides using recombinannt techniques. In addition, methods are provided for using such sequences to alter triacylglycerol levels in plants and to treat diseases associated with altered DAGAT activity or expression.

By this invention, a plant &bgr;-ketoacyl-CoA synthase condensing enzyme is provided free from intact cells of said plant and capable of catalyzing the production of very long chain fatty acid molecules. Also contemplated are constructs comprising the nucleic acid sequence and a heterologous DNA sequence not naturally associated with the condensing enzyme encoding sequences, and which provide for at least transcription of a plant condensing enzyme encoding sequence in a host cell. In this fashion very long chain fatty acid molecules may be produced in a plant cell. Included are methods of modifying the composition of very long chain fatty acid molecules in a plant cell.

Nucleic acid sequences encoding a plant cytoplasmic protein involved in fatty acyl-CoA metabolism

The invention provides diacylglycerol acyltransferase (DAGAT) proteins, wherein said proteins are active in the formation of triacylglycerol from fatty acyl and diacylglycerol substrates. In one aspect, DAGAT proteins have been isolated and have molecular weights of between approximately 36 and 37 kDa as measured by SDS-PAGE. The invention also provides novel DAGAT polynucleotide and polypeptide sequences and to methods of producing such polypeptides using recombinant techniques. In addition, methods are provided for using such sequences to alter triacylglycerol levels in plants and to treat diseases associated with altered DAGAT activity or expression.

By this invention, nucleic acid sequences encoding for fatty acyl-CoA: fatty alcohol acyltransferase (wax synthase) are provided, wherein said wax synthase is active in the formation of a wax ester from fatty alcohol and fatty acyl-CoA substrates. Of special interest is are nucleic acid sequences obtainable from a jojoba embryo wax synthase having an apparent molecular mass of approximately kD. Also considered are amino acid and nucleic acid sequences obtainable from wax synthase proteins and the use of such sequences to provide transgenic host cells capable of producing wax esters.

By this invention, nucleic acid sequences encoding for fatty acyl-CoA: fatty alcohol acyltransferase (wax synthase) are provided, wherein wax synthase is active in the formation of a wax ester from fatty alcohol and fatty acyl-CoA substrates. Of special interest is are nucleic acid sequences obtainable from a jojoba embryo wax synthase having an apparent molecular mass of approximately 33 kD. Also considered are amino acid and nucleic acid sequences obtainable from wax synthase proteins and the use of such sequences to provide transgenic host cells capable of producing wax esters.

By this invention, a plant cytoplasmic synthase protein is provided which is selected from the group consisting of a .beta.-ketoacyl-CoA synthase and a wax synthase, and is also free from intact cells of said plant and capable of catalyzing the production of very long chain fatty acid molecules. Also contemplated are constructs comprising the nucleic acid sequence and a heterologous DNA sequence not naturally associated with the plant cytoplasmic protein encoding sequences, and which provide for at least transcription of a plant cytoplasmic protein encoding sequence in a host cell. In this fashion very long chain fatty acid molecules may be produced in a host cell. Included are methods of modifying the composition of very long chain fatty acid molecules in a plant cell.

Nucleic acid sequences encoding a plant cytoplasmic protein involved in

By this invention, a partially purified fatty acyl-CoA: fatty alcohol acyltransferase (wax synthase) is provided, wherein said protein is active in the formation of a wax ester from fatty alcohol and fatty acyl substrates. Of special interest is a jojoba embryo wax synthase having an apparent molecular mass of approximately 57 kD. Also considered are amino acid and nucleic acid sequences obtainable from wax synthase proteins and the use of such sequences to provide transgenic host cells capable of producing wax esters.

Growing community of inventors

Woodland, CA, United States of America

Kathryn Dennis Lardizabal