BIOLOGICAL METHODS FOR PREPARING A FATTY DICARBOXYLIC ACID
The technology relates in part to biological methods for producing a fatty dicarboxylic acid and engineered microorganisms capable of such production.
1 - 159 . (canceled)
160 . A genetically modified yeast, comprising an active, modified endogenous acyl-coA Oxidase polypeptide, which yeast is capable of producing a diacid from a feedstock comprising one or more components from a vegetable oil.
161 . (canceled)
162 . The genetically modified yeast of claim 160 , wherein the acyl-coA Oxidase polypeptide comprises an amino acid modification at one or more amino acid positions chosen from 81, 82, 83, 84, 85, 86, 88, 93, 94, 95, 96, 98, 102, 284, 287, 290, 291, 292, 294, 295, 436, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462 and 463.
163 . The genetically modified yeast of claim 160 , wherein said modified acyl-coA Oxidase polypeptide comprises at least one amino acid substitution.
164 - 167 . (canceled)
168 . The genetically modified yeast of claim 162 , which comprises a genetic modification that reduces the activity of an enoyl coA isomerase polypeptide.
169 . The genetically modified yeast of claim 168 , wherein the genetic modification disrupts a polynucleotide that encodes the enoyl coA isomerase polypeptide.
170 . (canceled)
171 . The genetically modifies yeast of claim 168 , wherein the enoyl coA isomerase polypeptide is a polypeptide native to a yeast, wherein the yeast is a Candida spp. yeast.
172 . (canceled)
173 . The genetically modified yeast of claim 168 , wherein the diacid is a C4 to C24 diacid.
174 . The genetically modified yeast of claim 173 , wherein the diacid is a C10, C12, C14, C16, C18 or C20 diacid.
175 . (canceled)
176 . (canceled)
177 . The genetically modified yeast of claim 174 , wherein the diacid is the predominant diacid in a mixture of diacids.
178 . (canceled)
179 . The genetically modified yeast of claim 177 , wherein the feedstock comprises a plurality of fatty acids.
180 - 182 . (canceled)
183 . The genetically modified yeast of claim 168 , wherein the vegetable oil is from a plant chosen from palm, palm kernel, coconut, soy, safflower, canola, palm, palm kernel or combination thereof.
184 . A method for producing a diacid, comprising:
contacting a genetically modified yeast of claim 162 with a feedstock comprising one or more components from a vegetable oil capable of being converted by yeast to a diacid; and
culturing the yeast under conditions in which the diacid is produced from the feedstock.
185 . A diacid produced by the method of claim 184 .
186 . An expression product comprising a desired diacid, said expression product obtained by contacting a genetically modified yeast of claim 168 with a feedstock capable of being converted by the yeast to said desired diacid, and culturing the yeast under conditions in which the expression product comprising said diacid is produced from the feedstock.
187 . An expression product comprising at least one of sebacic acid and dodecanedioic acid, said product obtained by contacting a genetically modified yeast comprising an active, modified endogenous acyl-coA oxidase polypeptide with a feedstock capable of being converted by the yeast to at least one of sebacic acid and dodecanedioic acid; and culturing the yeast under conditions in which said expression product is produced.