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Title: Mutant fatty acid desaturase

Patent ·
OSTI ID:1174712

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.

Research Organization:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Organization:
USDOE
DOE Contract Number:
AC02-98CH10886
Assignee:
Brookhaven Science Associates LLC (Upton, NY)
Patent Number(s):
6,686,186
Application Number:
09/988,929
OSTI ID:
1174712
Country of Publication:
United States
Language:
English

References (8)

Engineering Δ 9 -16:0-Acyl Carrier Protein (ACP) Desaturase Specificity Based on Combinatorial Saturation Mutagenesis and Logical Redesign of the Castor Δ 9 -18:0-ACP Desaturase journal April 2001
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
.beta.-Hydroxydecanoyl thioester dehydrase does not catalyze a rate-limiting step in Escherichia coli unsaturated fatty acid synthesis journal December 1983
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

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