Plant C18:1 preferring thioesterases

• US 6,028,247 A
• Filed: 06/06/1995
• Issued: 02/22/2000
• Est. Priority Date: 04/26/1990
• Status: Expired due to Term

• Current Assignee: Monsanto Company (Bayer AG)
• Original Assignee: Calgene Incorporated (Pharmacia Corp.)
• Inventors: Thompson, Gregory A., Voelker, Toni Alois, Davies, Hue Maelor
• Primary Examiner(s): Robinson, Douglas W.
• Assistant Examiner(s): Nelson, Amy J.
• Application Number:
  US08/470,204
• Time in Patent Office: 1,722 Days
• Field of Search: 435/320.1, 435/172.3, 435/240.4, 435/252.3, 435/419, 800/205, 800/250, 800/DIG. 15, 536/23.2, 536/23.6
• US Class Current: 800/281
• CPC Class Codes: A23D 9/00 Other edible oils or fats, e.g. shortenings, cooking oils
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)
• CPC Class Codes: C12P 7/6418 by hydrolysis of fatty acid esters
• CPC Class Codes: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil

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

1. A recombinant DNA construct comprising a plant C18:

1 preferring acyl-ACP thioesterase encoding sequence, wherein said C18;

1 preferring thioesterase is from safflower or Brassica.

16. A construct according to any one of claims 1, 2 or 5, wherein said C18:

1 preferring thioesterase sequence comprises thioesterase encoding sequence provided in any one of SEQ ID NO;

44, SEQ ID NO;

45 and SEQ ID NO;

48.

17. A construct according to any one of claims 1, 2 or 5, wherein said C18:

1 preferring thioesterase comprises the amino acid sequence encoded by nucleotides 173-1339 of SEQ ID NO;

44 or nucleotides 94-1248 of SEQ ID NO;

45.

2. A recombinant DNA construct comprising as operably linked components in the 5'"'"' to 3'"'"' direction of transcription, a transcriptional initiation region functional in a host plant or bacterial cell and a plant C18:

1 preferring acyl-ACP thioesterase nucleic acid sequence, wherein said transcriptional initiation region is not naturally linked to said thioesterase sequence, and wherein said C18;

1 preferring thioesterase is from safflower or Brassica.

3. The construct of claim 2 further comprising, 3'"'"' to said thioesterase nucleic acid sequence, a transcriptional termination region functional in a host cell.

4. The construct of claim 2, wherein said thioesterase nucleic acid sequence is oriented for transcription of a sense sequence.

5. A DNA construct comprising an expression cassette capable of producing a plant thioesterase in a plant cell comprising as operably linked components in the 5'"'"' to 3'"'"' direction of transcription, a transcriptional initiation regulatory region functional in said plant cell, a translational initiation regulatory region functional in said plant cell, a sense DNA sequence encoding a biologically active plant C18:

1 preferring acyl-ACP thioesterase, and a transcriptional termination regulatory region functional in said plant cell, wherein at least one of said regulatory regions is not naturally linked to said sense DNA sequence, and wherein said C18;

1 preferring thioesterase is from safflower or Brassica.

6. The construct of claim 5 wherein said C18:

1 preferring acyl-ACP thioesterase also demonstrates activity towards C16;

0 and C18;

0 acyl-ACP substrates.

7. The construct of claim 5 wherein said transcriptional initiation region is functional in a plant seed cell.

8. The construct of claim 7 wherein said transcriptional initiation region is from a gene selected from the group consisting of napin, Bce4, phaseolin and acyl-carrier protein.

9. The construct of claim 5, wherein said construct further comprises a marker for detection of cells comprising said marker.

10. A plant cell comprising a construct according to claim 5.

11. A plant comprising a plant cell according to claim 10.

12. A method of modifying the fatty acid composition in a plant cell comprising:

growing a plant cell having integrated in its genome a DNA construct, said construct comprising in the 5'"'"' to 3'"'"' direction of transcription, a transcriptional regulatory region functional in said plant cell and a plant C18;

1 preferring acyl-ACP thioesterase encoding sequence, under conditions which will permit the transcription of said plant C18;

1 preferring acyl-ACP thioesterase.

13. The method of claim 12 wherein said C18:

1 preferring acyl-ACP thioesterase is from safflower or Brassica.

14. The method according to claim 13 wherein said encoding sequence comprises C18:

1 preferring thioesterase encoding sequence provided in any one of SEQ ID NO;

44, SEQ ID NO;

45 and SEQ ID NO;

48.

15. The method according to claim 13 wherein said C18:

1 preferring thioesterase comprises the amino acid sequence encoded by nucleotides 173-1339 of SEQ ID NO;

44 or nucleotides 94-1248 of SEQ ID NO;

45.