Calgene Corporation

Other

Roussel Uclaf

Toni Alois Voelker

Huw Maelor Davies

Danielle Lando

Charles J Van Assche

Monica Gervais

Jean Michel Bruneau

Jean C Kridl

Gregory A Thompson

Deborah J Hawkins

Deborah Knutzon

Aubrey Jones

Ling Yuan

Hue Maelor Davies

This invention relates to plant thioesterases, means to identify such proteins, amino acid and nucleic acid sequences associated with such protein, methods to obtain, make and/or use such plant thioesterases. Also, by this invention, the existence of a heretofore unproven factor critical to the biosynthesis of medium-chain fatty acids in plants is demonstrated.

Plant C18:1 preferring thioesterases

Sucrose phosphate synthase (SPS), a process for its preparation, its DNA, and utilization of DNA encoding SPS to modify the expression of SPS in the plant cells are provided. Methods and compositions for producing SPS protein and DNA encoding SPS from a number of sources are provided. Also provided are transgenes comprising DNA encoding SPS and methods for the production of transgenic plant cells and plants which have extra SPS activity, and thus an increased capacity to synthesize sucrose. Transgenic plant cells and plants with the additional SPS activity exhibit altered carbon partitioning, growth and development, and yield under a variety of conditions.

Modification of sucrose phosphate synthase in plants

A cDNA coating for a protein having saccharose phosphate synthetase activity and a vector permitting the expression of a protein having saccharose phosphate synthetase activity under the control of a promoter directing the said expression.

Saccharose phosphate synthetase (SPS), a preparation method and CDNA

By this invention, further properties and uses of plant medium chain thioesterases are provided. In particular, it is now seen that plant seed which would not normally contain medium-chain fatty acid, either as free fatty acids or incorporated into triglyceride molecules, can be found to contain such medium-chain fatty acids. By seed which would not normally contain medium-chain fatty acid is meant seed which contains less than 0.1 mole percent of a given medium-chain fatty acid in total fatty acids. Thus, any plant seed containing a minimum of 1.0 mole percent of a given medium-chain fatty acid in total fatty acids is significantly modified. In one embodiment, this invention relates to plant seed and oil derived from that seed, which normally do not contain laurate, but now are found to contain laurate.

Plant thioesterases and use for modification of fatty acid composition

This invention relates to plant thioesterases, specifically plant acyl-ACP thioesterases having substantial activity on palmitoyl-ACP substrates. DNA constructs useful for the expression of a plant palmitoyl-ACP thioesterase in a plant seed cell are described. Such constructs will contain a DNA sequence encoding the plant palmitoyl-ACP thioesterase of interest under the control of regulatory elements capable of preferentially directing the expression of the plant palmitoyl-ACP thioesterase in seed tissue, as compared with other plant tissues, when such a construct is expressed in a transgenic plant. This invention also relates to methods of using a DNA sequence encoding a plant palmitoyl-ACP thioesterase for the modification of the proportion of free fatty acids produced in a plant seed cell. Plant palmitoyl-ACP thioesterase sequences exemplified herein include Cuphea, leek, mango and elm. Transgenic plants having increased levels of C16:0 fatty acids in their seeds as the result of expression of these palmitoyl-ACP thioesterase sequences are also provided.

Plant acyl-ACP thioesterase sequences

A protein having sucrose phosphate synthetase (SPS) activity is isolated from plants, preferably maize. The protein has a molecular weight of 110-130 dK and contains at least one peptide selected from Thv Trp Ile Lys, Try Val Val Glu Leu Ala Arg, Ser Met Pro Pro Ile Trp Ala Glu Val Met Arg, Leu Arg Pro Asp Gln Asp Try Leu Met His Ile Ser His Arg and Trp Ser His Asp Gly Ala Arg. Isolation is carried out by obtaining an extract from the plant by grinding, centrifugation and filtration; enriching the extract in SPS protein by precipitation in an appropriate solvent such as polyethylene glycol, centrifugation and solubilization of the precipitate obtained in a buffer solution; subjecting the protein thus obtained to low pressure anion exchange chromatography, chromatography on heparin Sepharose and high pressure anion exchange chromatography; and purifying the active fractions obtained by passage through two high pressure chromatography columns. Hybridomas and monoclonal antibodies are prepared from an antigen resulting from high pressure anion exchange chromatography above, antibodies directed specifically against SPS are selected and the antibodies are used to purify the SPS obtained previously. Complementary DNA coding for the SPS is prepared and used to modify expression of the SPS in plant cells.

Sucrose phosphate synthetase isolated from maize

C8 and C10 medium-chain thioesterases in plants

Sucrose phosphate synthase (SPS), its process for preparation, its cDNA, and utilization of cDNA to modify the expression of SPS in the plant cells are provided.

Sucrose phosphate synthase (SPS), its process for preparation its cDNA,

By this invention, methods to produce C14 fatty acids in plant seed oils are provided. In a first embodiment, this invention relates to particular C14 preferring acyl-ACP thioesterase sequences from Cuphea palustris, camphor and nutmeg, and to DNA constructs for the expression of these thioesterases in host cells for production of C14 fatty acids. Other aspects of this invention relate to methods for using other plant medium-chain thioesterases or medium-chain thioesterases from non-plant sources to provide C14 fatty acids in plant cells. In this regard, the production of C14 fatty acids in plant cells as the result of expression from an elm medium chain acyl-ACP thioesterase and a bacterial luxD gene is provided.

Production of myristate in plant cells

Plant thioesterases

By this invention, further properties and uses of medium-chain thioesterases in plants are provided, including methods of using medium-chain thioesterases from non-plant sources to provide medium-chain fatty acids in plant cells.

Medium-chain thioesterases in plants

Plant thiosterases

This invention relates to a novel class of plant enzymes, thioesterases having preferential hydrolase activity toward one or more medium-chain acyl-ACP substrates. Purification, especially the removal of substantially all other plant proteins, and use of the plant thioesterase is provided, including the use of the protein as a tool in gene isolation for biotechnological applications and a description of various uses for such nucleic acid sequences thereto.

Growing community of inventors

Davis, CA, United States of America

Toni Alois Voelker