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Title: Mutant fatty acid desaturase

Abstract

The present invention relates to a method for producing mutants of a fatty acid desaturase having a substantially increased activity towards fatty acid substrates with chains containing fewer than 18 carbons relative to an unmutagenized precursor desaturase having an 18 carbon atom chain length substrate specificity. The method involves inducing one or more mutations in the nucleic acid sequence encoding the precursor desaturase, transforming the mutated sequence into an unsaturated fatty acid auxotroph cell such as MH13 E. coli, culturing the cells in the absence of supplemental unsaturated fatty acids, thereby selecting for recipient cells which have received and which express a mutant fatty acid desaturase with an elevated specificity for fatty acid substrates having chain lengths of less than 18 carbon atoms. A variety of mutants having 16 or fewer carbon atom chain length substrate specificities are produced by this method. Mutant desaturases produced by this method can be introduced via expression vectors into prokaryotic and eukaryotic cells and can also be used in the production of transgenic plants which may be used to produce specific fatty acid products.

Inventors:
;
Issue Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1174712
Patent Number(s):
6686186
Application Number:
09/988,929
Assignee:
Brookhaven Science Associates LLC (Upton, NY)
Patent Classifications (CPCs):
C - CHEMISTRY C12 - BIOCHEMISTRY C12N - MICROORGANISMS OR ENZYMES
DOE Contract Number:  
AC02-98CH10886
Resource Type:
Patent
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Shanklin, John, and Cahoon, Edgar B. Mutant fatty acid desaturase. United States: N. p., 2004. Web.
Shanklin, John, & Cahoon, Edgar B. Mutant fatty acid desaturase. United States.
Shanklin, John, and Cahoon, Edgar B. Tue . "Mutant fatty acid desaturase". United States. https://www.osti.gov/servlets/purl/1174712.
@article{osti_1174712,
title = {Mutant fatty acid desaturase},
author = {Shanklin, John and Cahoon, Edgar B.},
abstractNote = {The present invention relates to a method for producing mutants of a fatty acid desaturase having a substantially increased activity towards fatty acid substrates with chains containing fewer than 18 carbons relative to an unmutagenized precursor desaturase having an 18 carbon atom chain length substrate specificity. The method involves inducing one or more mutations in the nucleic acid sequence encoding the precursor desaturase, transforming the mutated sequence into an unsaturated fatty acid auxotroph cell such as MH13 E. coli, culturing the cells in the absence of supplemental unsaturated fatty acids, thereby selecting for recipient cells which have received and which express a mutant fatty acid desaturase with an elevated specificity for fatty acid substrates having chain lengths of less than 18 carbon atoms. A variety of mutants having 16 or fewer carbon atom chain length substrate specificities are produced by this method. Mutant desaturases produced by this method can be introduced via expression vectors into prokaryotic and eukaryotic cells and can also be used in the production of transgenic plants which may be used to produce specific fatty acid products.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {2004},
month = {2}
}

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Works referenced in this record:

Redesign of soluble fatty acid desaturases from plants for altered substrate specificity and double bond position
journal, May 1997


Primary structures of the precursor and mature forms of stearoyl-acyl carrier protein desaturase from safflower embryos and requirement of ferredoxin for enzyme activity.
journal, March 1991


Desaturation and Related Modifications of Fatty Acids
journal, June 1998


Combinatorial protein design: strategies for screening protein libraries
journal, August 1997


Directed evolution of a thermostable esterase
journal, October 1998


An efficient method for generating proteins with altered enzymatic properties: application to beta-lactamase.
journal, December 1989