Engineering plant thioesterases for altered substrate specificit

Details Engineering plant thioesterases for altered substrate specificit Engineering plant thioesterases for altered substrate specificit

1995-05-15

1999-09-21

Wax, Robert A.

Chemistry: molecular biology and microbiology

Treatment of micro-organisms or enzymes with electrical or...

Modification of viruses

4351723, 435196, 435 691, 4353201, 4352523, 43525233, 435410, 536 231, 536 232, 536 236, C12N 1500, C07H 2104

Patent

active

059553294

ABSTRACT:
Methods of altering substrate specificity of plant acyl-ACP thioesterases, and engineered plant acyl-ACP thioesterases so produced are provided. The C-terminal two thirds portion of plant thioesterases is identified as desirable for such modifications. DNA sequences and constructs for expression of engineered thioesterases, as well as the novel thioesterases produced therefrom are also provided. Such DNA sequences may be used for expression of the engineered thioesterases in host cells, particularly seed cells of oilseed crop plants, for the modification of fatty acid composition. A C12 preferring plant acyl-ACP thioesterase described herein may be altered to obtain a plant thioesterase having approximately equal activity on C14 and C12 substrates. Further modification of the C12 enzyme yields a thioesterase having greater activity on C14 as compared to C12 substrates. Also provided in the instant invention are novel plant acyl-ACP thioesterase sequences from Cuphea palustris and mangosteen (Garcinia mangifera), which demonstrate desirable thioesterase activity profiles for plant genetic engineering applications.

REFERENCES:
patent: 5344771 (1994-09-01), Davies et al.
patent: 5455167 (1995-10-01), Voelker et al.
patent: 5512482 (1996-04-01), Voelker et al.
Jenks, W. P. "Catalysis in Chemistry and Enzymology" Dover Edition, Dover Publication, New York, pp. 615-623, 1987.
Armstrong, R. N. "Structure-Function Relationship in enzymic catalysis. Can chimeric enzymes contribute?" Chem. Rev. 90, 1309-1325, 1990.
Dormann et al. (B) "Cloning and expression in Escherichia coli of a novel thioesterase from Arabidopsis thaliana . . . " Arch. Biochem. Biophys. 316(1), 612-618, Jan. 10, 1995.
Dormann et al. (A) "Cloning and expression in Escherichia coli of cDNA coding for the oleoyl-acyl carrier protein . . . " Biochemica et Biophysica Acta 1212, 134-136, 1994.
Knutzon et al. "Isolation and characterization of two safflower oleoyl-acyl carrier protein thioesterase cDNA clones" Plant Physiol. 100, 1751-1758, 1992.
Loader et al. "Isolation and characterization of two Brassica napus embryo acy-ACP thioesterase cDNA clones" Plant Mol. Biol. 23, 769-778, Nov. 1993.
Voelker et al. "Fatty acid biosynthesis redirected to medium chains in transgenic oilseed plants" Science 257, 72-74, Jul. 3, 1992.
Yuan, et al., "The catalytic cysteine and histidine in the plant acyl-acyl carrier protein thioesterases" Journal of Biological Chemistry 271 vol. 7:3417-3419 (1996).
Yuan, et al., "Modification of the substrate specificity of an acyl-acyl carrier protein thioesterase by protein engineering", Proceedings of the National Academy of Sciences of the United States of America 92 vol. 23:10639-10643 (1995).
Topfer, R, et al., "Modification of plant lipid synthesis", Science vol. 268:681-686 (1995).
Tai, M-H et al., "Roles of Ser-101 ASP-236 and HIS-237 in Catalysis of Thioesterase II and of the C-terminal region of the Enzyme in its interaction with Fatty Acid Synthase". Proceeds of the National Academy of Science USA 90 vol. 5:1852-1856 (1993).
Kim, et al., "Chimeric restriction endonuclease" Proc. Natl. Acad. Sci. USA Vo. 91, pp. 883-887, Feb. 1994.
Hjelmstad, et al., "Chimeric Enzymes" The Journal of Biological Chemistry, vol. 269, No. 33, pp. 20995-21002 (1994).
Jones, et al., "Palmitoyl-Acyl Carrier Protein (ACP) Thioesterase and the Evolutionary Origin of Plant Acyl-ACP Thioesterases" The Plant Cell, vol. 7, pp. 359-371, Mar. 1995.
Lawson, et al., "Structure of a Myristoyl-ACP-Specific Thioesterase from Vibrio harveyi", Biochemistry, 1994, 33 9382-9388.

Affiliated with

Also associated with

Rating

Engineering plant thioesterases for altered substrate specificit does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Engineering plant thioesterases for altered substrate specificit, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Engineering plant thioesterases for altered substrate specificit will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-79464