# US 5,344,771 A · Plant thiosterases

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- Number: US 5,344,771 A
- Document ID: US5344771A
- Status: Granted
- Kind: A
- Application number: 07/662,007
- Filed: 1991-02-27
- Granted: 1994-09-06
- Earliest claimed: 1990-04-26
- Assignees: Calgene, Inc.
- Inventors: Huw M. Davies; Toni A. Voelker
- Web page: https://patentgrep.com/patent/US5344771A

## CPC

- C12P 7/6418: by hydrolysis of fatty acid esters
- C07K 14/415: from plants
- C12N 9/16: acting on ester bonds (3.1)
- C12N 15/8247: {involving modified lipid metabolism, e.g. seed oil composition}
- C12P 7/6463: obtained from glyceride producing microorganisms, e.g. single cell oil
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

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.

## Claims

### Claim 1

A method of producing C12:0 fatty acids in a Brassica seed cell, said method comprising:
growing a Brassica plant having integrated into its genome a DNA construct, said construct comprising in the 5' to 3' direction of transcription, a transcriptional regulatory region functional in said Brassica seed cell, a translational regulatory region functional in said Brassica seed cell, a plant transit peptide encoding sequence, a DNA sequence encoding an Umbellularia californica C12:0 preferring acyl-ACP thioesterase functional in said Brassica seed cell, and a transcriptional termination region functional in said Brassica seed cell.

### Claim 2 (depends on 1)

The method of claim 1 wherein said transcriptional regulatory region is a CaMV double 35S promoter.

### Claim 3 (depends on 1)

The method of claim 1 wherein said transcriptional regulatory region is from a gene preferentially expressed in a plant seed tissue.

### Claim 4 (depends on 1)

The method of claim 1 wherein said transcriptional regulatory region is from a gene selected from the group consisting of napin, phaseolin and Bce4.

### Claim 5 (depends on 1)

The method of claim 1 wherein said Brassica plant is B. napus.

### Claim 6 (depends on 1)

A transgenic Brassica seed cell comprising an increased percentage of C12:0 fatty acids as compared to the percentage of C12:0 fatty acids in a non-transgenic Brassica seed cell, wherein said increased percentage of C12:0 fatty acids is produced according to the method of claim 1.