Tailored Oils Produced From Recombinant Heterotrophic Microorganisms

• US 20110294174A1
• Filed: 05/27/2011
• Published: 12/01/2011
• Est. Priority Date: 05/28/2010
• Status: Active Grant

• Current Assignee: Terravia Holdings, Inc. (Corbion NV)
• Original Assignee: Solazyme Inc.
• Inventors: Rudenko, George, Somanchi, Aravind, Franklin, Scott, Rakitsky, Walt, Moseley, Jeffrey L., Wee, Janice
• Granted Patent: US 8,592,188 B2
• Time in Patent Office: Days
• US Class Current: 435/134
• CPC Class Codes: A23D 7/00 Edible oil or fat compositions containing an aqueous phase, e.g. margarines
• CPC Class Codes: A23D 9/00 Other edible oils or fats, e.g. shortenings, cooking oils
• CPC Class Codes: A23K 20/158 Fatty acids; Fats; Products containing oils or fats
• CPC Class Codes: C11B 1/00 Production of fats or fatty oils from raw materials
• CPC Class Codes: C11C 3/00 Fats, oils, or fatty acids by chemical modification of fats, oils, or fatty acids obtained therefrom sulfonated fats or oils C07C309/62; factice C08H; drying oils C09F
• CPC Class Codes: C12N 15/67 General methods for enhancing the expression
• CPC Class Codes: C12N 15/79 Vectors or expression systems specially adapted for eukaryotic hosts
• CPC Class Codes: C12N 15/8216 Methods for controlling, regulating or enhancing expression of transgenes in plant cells
• CPC Class Codes: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)
• CPC Class Codes: C12N 9/2431 Beta-fructofuranosidase (3.2.1.26), i.e. invertase
• CPC Class Codes: C12N 9/2465 acting on alpha-galactose-glycoside bonds, e.g. alpha-galactosidase (3.2.1.22)
• CPC Class Codes: C12P 7/64 Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
• CPC Class Codes: C12P 7/6445 Glycerides
• CPC Class Codes: C12P 7/6463 obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12Y 114/19001 Stearoyl-CoA 9-desaturase (1.14.19.1), i.e. DELTA9-desaturase
• CPC Class Codes: C12Y 301/02 Thioester hydrolases (3.1.2)
• CPC Class Codes: C12Y 301/02014 Oleoyl-[acyl-carrier-protein] hydrolase (3.1.2.14), i.e. ACP-thioesterase
• CPC Class Codes: C12Y 302/01022 Alpha-galactosidase (3.2.1.22)
• CPC Class Codes: C12Y 302/01026 Beta-fructofuranosidase (3.2.1.26), i.e. invertase
• CPC Class Codes: Y02E 50/10 : Biofuels, e.g. bio-diesel
• CPC Class Codes: Y02P 30/20 : using bio-feedstock
• CPC Class Codes: Y02T 50/678 : Aviation using fuels of non-fossil origin

Abstract

Methods and compositions for the production of oil, fuels, oleochemicals, and other compounds in recombinant microorganisms are provided, including oil-bearing microorganisms and methods of low cost cultivation of such microorganisms. Microalgal cells containing exogenous genes encoding, for example, a lipase, a sucrose transporter, a sucrose invertase, a fructokinase, a polysaccharide-degrading enzyme, a keto acyl-ACP synthase enzyme, a fatty acyl-ACP thioesterase, a fatty acyl-CoA/aldehyde reductase, a fatty acyl-CoA reductase, a fatty aldehyde reductase, a fatty aldehyde decarbonylase, and/or an acyl carrier protein are useful in manufacturing transportation fuels such as renewable diesel, biodiesel, and renewable jet fuel, as well as oleochemicals such as functional fluids, surfactants, soaps and lubricants.

Claims

1. An oleaginous microbial cell comprising triglyceride oil, wherein the fatty acid profile of the triglyceride oil is selected from the group consisting of at least about 1% C8:

0, at least about 1% C10;

0, at least about 1% C12;

0, at least about 2% C14;

0, at least about 30% C16;

0, at least about 5% C18;

0, at least about 60% C18;

1, less than about 7% C18;

2, and at least about 35% saturated fatty acids.

2. The oleaginous microbial cell of claim 1, wherein said oleaginous microbial cell comprises an exogenous gene, and wherein optionally, an endogenous desaturase of said oleaginous microbial cell has been inactivated or mutated to have less enzymatic activity.

3. The oleaginous microbial cell of claim 2, wherein the fatty acid profile of the triglyceride oil is similar to the triglyceride oil profile of a naturally occurring oil.

4. The oleaginous microbial cell of claim 3, wherein the naturally occurring oil is selected from the group consisting of cocoa butter, coconut oil, palm oil, palm kernel oil, shea butter, beef tallow and lard.

5. The oleaginous microbial cell of claim 2, wherein the fatty acid profile of the triglyceride oil comprises a profile selected from the group consisting of, the total combined amounts of C8:

0 and C10;

0 is at least about 10%, the total combined amount of C10;

0, C12;

0, and C14;

0 is at least about 50%, the total combined amount of C16;

0, C18;

0 and C18;

1 is at least about 60%, the total combined amount of C18;

0, C18;

1 and C18;

2 is at least about 60%, the total combined amount of C14;

0, C16;

0, C18;

0 and C18;

1 is at least about 60%, and the total combined amount of C18;

1 and C18;

2 is less than about 30%.

6. The oleaginous microbial cell of claim 1, wherein the fatty acid profile of the triglyceride oil comprises a ratio of fatty acids selected from the group consisting of C8:

0 to C10;

0 ratio of at least about 5 to 1, C10;

0 to C12;

0 ratio of at least about 6 to 1, C12;

0 to C14;

0 ratio of at least about 5 to 1, C14;

0 to C12;

0 ratio of at least about 7;

1, and C14;

0 to C16;

0 ratio of at least about 1 to 2.

7. The oleaginous microbial cell of claim 2, wherein said endogenous desaturase is selected from the group consisting of stearoyl ACP desaturase and delta 12 fatty acid desaturase.

8. The oleaginous microbial cell of claim 7, wherein said exogenous gene is selected from the group consisting of a gene encoding an acyl-ACP thioesterase.

9. The oleaginous microbial cell of claim 8, wherein said exogenous gene encodes an acyl-ACP thioesterase selected from the group consisting of Umbellularia californica fatty acyl-ACP thioesterase (GenBank #AAC49001);

Cinnamomum camphora fatty acyl-ACP thioesterase (GenBank #Q39473);

Umbellularia californica fatty acyl-ACP thioesterase (GenBank #Q41635);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank #AAB71729);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank #AAB71730);

Elaeis guineensis fatty acyl-ACP thioesterase (GenBank #ABD83939);

Elaeis guineensis fatty acyl-ACP thioesterase (GenBank #AAD42220);

Populus tomentosa fatty acyl-ACP thioesterase (GenBank #ABC47311);

Arabidopsis thaliana fatty acyl-ACP thioesterase (GenBank #NP_—

172327);

Arabidopsis thaliana fatty acyl-ACP thioesterase (GenBank #CAA85387);

Arabidopsis thaliana fatty acyl-ACP thioesterase (GenBank #CAA85388);

Gossypium hirsutum fatty acyl-ACP thioesterase (GenBank #Q9SQI3);

Cuphea lanceolata fatty acyl-ACP thioesterase (GenBank #CAA54060);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #AAC72882);

Cuphea calophylla subsp. mesostemon fatty acyl-ACP thioesterase (GenBank #ABB71581);

Cuphea lanceolata fatty acyl-ACP thioesterase (GenBank #CAC19933);

Elaeis guineensis fatty acyl-ACP thioesterase (GenBank #AAL15645);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #Q39513);

Gossypium hirsutum fatty acyl-ACP thioesterase (GenBank #AAD01982);

Vitis vinifera fatty acyl-ACP thioesterase (GenBank #CAN81819);

Garcinia mangostana fatty acyl-ACP thioesterase (GenBank #AAB51525);

Brassica juncea fatty acyl-ACP thioesterase (GenBank #ABI18986);

Madhuca longifolia fatty acyl-ACP thioesterase (GenBank #AAX51637);

Brassica napus fatty acyl-ACP thioesterase (GenBank #ABH11710);

Oryza sativa (indica cultivar-group) fatty acyl-ACP thioesterase (GenBank #EAY86877);

Oryza sativa (japonica cultivar-group) fatty acyl-ACP thioesterase (GenBank #NP_—

001068400);

Oryza sativa (indica cultivar-group) fatty acyl-ACP thioesterase (GenBank #EAY99617);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #AAC49269);

Ulmus Americana fatty acyl-ACP thioesterase (GenBank #AAB71731);

Cuphea lanceolata fatty acyl-ACP thioesterase (GenBank #CAB60830);

Cuphea palustris fatty acyl-ACP thioesterase (GenBank #AAC49180);

Iris germanica fatty acyl-ACP thioesterase (GenBank #AAG43858);

Iris germanica fatty acyl-ACP thioesterase (GenBank #AAG43858.1);

Cuphea palustris fatty acyl-ACP thioesterase (GenBank #AAC49179);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank# AAB71729);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank# AAB717291.1);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #U39834);

Umbelluaria californica fatty acyl-ACP thioesterase (GenBank # M94159);

Cinnamomum camphora fatty acyl-ACP thioesterase (GenBank #U31813);

Cuphea wrightii fatty acyl-ACOP thioesterase (GenBank #U56103); and

Ricinus communis fatty acyl-ACP thioesterase (GenBank #ABS30422).

10. The oleaginous microbial cell of claim 2, wherein said oleaginous microbial cell further comprises a gene encoding a sucrose invertase.

11. The oleaginous microbial cell of claim 2, wherein said oleaginous microbial cell is a cell of a microalgal genus or species selected from Achnanthes orientalis, Agmenellum, Amphiprora hyaline, Amphora coffeiformis, Amphora coffeiformis linea, Amphora coffeiformis punctata, Amphora coffeiformis taylori, Amphora coffeiformis tenuis, Amphora delicatissima, Amphora delicatissima capitata, Amphora sp., Anabaena, Ankistrodesmus, Ankistrodesmus falcatus, Boekelovia hooglandii, Borodinella sp., Botryococcus braunii, Botryococcus sudeticus, Carteria, Chaetoceros gracilis, Chaetoceros muelleri, Chaetoceros muelleri subsalsum, Chaetoceros sp., Chlorella anitrata, Chlorella Antarctica, Chlorella aureoviridis, Chlorella candida, Chlorella capsulate, Chlorella desiccate, Chlorella ellipsoidea, Chlorella emersonii, Chlorella fusca, Chlorella fusca var. vacuolata, Chlorella glucotropha, Chlorella infusionum, Chlorella infusionum var. actophila, Chlorella infusionum var. auxenophila, Chlorella kessleri, Chlorella lobophora (strain SAG 37.88), Chlorella luteoviridis, Chlorella luteoviridis var. aureoviridis, Chlorella luteoviridis var. lutescens, Chlorella miniata, Chlorella minutissima, Chlorella mutabilis, Chlorella nocturna, Chlorella parva, Chlorella photophila, Chlorella pringsheimii, Chlorella protothecoides (including any of UTEX strains 1806, 411, 264, 256, 255, 250, 249, 31, 29, 25, and CCAP strains 211/17 and 211/8d), Chlorella protothecoides var. acidicola, Chlorella regularis, Chlorella regularis var. minima, Chlorella regularis var. umbricata, Chlorella reisiglii, Chlorella saccharophila, Chlorella saccharophila var. ellipsoidea, Chlorella salina, Chlorella simplex, Chlorella sorokiniana, Chlorella sp., Chlorella sphaerica, Chlorella stigmatophora, Chlorella vanniellii, Chlorella vulgaris, Chlorella vulgaris, Chlorella vulgaris f. tertia, Chlorella vulgaris var. autotrophica, Chlorella vulgaris var. viridis, Chlorella vulgaris var. vulgaris, Chlorella vulgaris var. vulgaris f. tertia, Chlorella vulgaris var. vulgaris f. viridis, Chlorella xanthella, Chlorella zofingiensis, Chlorella trebouxioides, Chlorella vulgaris, Chlorococcum infusionum, Chlorococcum sp., Chlorogonium, Chroomonas sp., Chrysosphaera sp., Cricosphaera sp., Cryptomonas sp., Cyclotella cryptica, Cyclotella meneghiniana, Cyclotella sp., Dunaliella sp., Dunaliella bardawil, Dunaliella bioculata, Dunaliella granulate, Dunaliella maritime, Dunaliella minuta, Dunaliella parva, Dunaliella peircei, Dunaliella primolecta, Dunaliella salina, Dunaliella terricola, Dunaliella tertiolecta, Dunaliella viridis, Dunaliella tertiolecta, Eremosphaera viridis, Eremosphaera sp., Ellipsoidon sp., Euglena, Franceia sp., Fragilaria crotonensis, Fragilaria sp., Gleocapsa sp., Gloeothamnion sp., Hymenomonas sp., Isochrysis aff. galbana, Isochrysis galbana, Lepocinclis, Micractinium, Micractinium (UTEX LB 2614), Monoraphidium minutum, Monoraphidium sp., Nannochloris sp., Nannochloropsis salina, Nannochloropsis sp., Navicula acceptata, Navicula biskanterae, Navicula pseudotenelloides, Navicula pelliculosa, Navicula saprophila, Navicula sp., Nephrochloris sp., Nephroselmis sp., Nitschia communis, Nitzschia alexandrina, Nitzschia communis, Nitzschia dissipata, Nitzschia frustulum, Nitzschia hantzschiana, Nitzschia inconspicua, Nitzschia intermedia, Nitzschia microcephala, Nitzschia pusilla, Nitzschia pusilla elliptica, Nitzschia pusilla monoensis, Nitzschia quadrangular, Nitzschia sp., Ochromonas sp., Oocystis parva, Oocystis pusilla, Oocystis sp., Oscillatoria limnetica, Oscillatoria sp., Oscillatoria subbrevis, Pascheria acidophila, Pavlova sp., Phagus, Phormidium, Platymonas sp., Pleurochrysis carterae, Pleurochrysis dentate, Pleurochrysis sp., Prototheca wickerhamii, Prototheca stagnora, Prototheca portoricensis, Prototheca moriformis, Prototheca zopfii, Pyramimonas sp., Pyrobotrys, Sarcinoid chrysophyte, Scenedesmus armatus, Spirogyra, Spirulina platensis, Stichococcus sp., Synechococcus sp., Tetraedron, Tetraselmis sp., Tetraselmis suecica, Thalassiosira weissflogii, and Viridiella fridericiana.

12. The oleaginous microbial cell of claim 11, wherein said oleaginous microbial cell is a cell of the genus Prototheca.

13. The oleaginous microbial cell of claim 12, wherein said oleaginous microbial cell is a cell of the genus Prototheca moriformis.

14. The oleaginous microbial cell of claim 2, wherein said oleaginous microbial cell is an oleaginous yeast cell.

15. The oleaginous microbial cell of claim 2, wherein said oleaginous microbial cell is an oleaginous bacterial cell.

16. The oleaginous microbial cell of claim 4, wherein said naturally occurring oil is cocoa butter and wherein said exogenous gene comprises a Carthamus tinctorus thioesterase gene.

17. The oleaginous microbial cell of claim 4, wherein said naturally occurring oil is coconut oil.

18. The oleaginous microbial cell of claim 4, wherein said naturally occurring oil is palm oil and wherein said exogenous gene comprises a Elaeis guiniensis thioesterase gene, a Cuphea hookeriana thioesterase gene, a combination of a Cuphea hookeriana KAS IV gene and a Cuphea wrightii FATB2 gene, or a construct designed to disrupt an endogenous KAS II gene.

19. The oleaginous microbial cell of claim 4, wherein said naturally occurring oil is palm kernel oil and wherein said exogenous gene comprises a combination of a Cuphea wrightii FATB2 gene and a construct designed to disrupt an endogenous SAD2B gene.

20. The oleaginous microbial cell of claim 4, wherein said naturally occurring oil is shea butter.

21. The oleaginous microbial cell of claim 4, wherein said naturally occurring oil is beef tallow.

22. The oleaginous microbial cell of claim 4, wherein said naturally occurring oil is lard and wherein said exogenous gene comprises a combination of U. californica thioesterase gene and a construct designed to disrupt an endogenous SAD2B gene, a combination of a Garcinia mangostana thioesterase gene and a construct designed to disrupt an endogenous SAD2B gene, a Brassica napus thioesterase gene, or a Cuphea hookeriana thioesterase gene.

64. A method of making an oil-based product, the method comprising the steps of:

a. subjecting the oleaginous microbial triglyceride oil composition of claim 21 to at least one chemical reaction selected from the group consisting of;

saponification;

metathesis;

acid hydrolysis;

alkaline hydrolysis;

enzymatic hydrolysis;

catalytic hydrolysis;

hot-compressed water hydrolysis;

a catalytic hydrolysis reaction wherein the lipid is split into glycerol and fatty acids;

an amination reaction to produce fatty nitrogen compounds;

an ozonolysis reaction to produce mono- and dibasic-acids;

a triglyceride splitting reaction selected from the group consisting of enzymatic splitting and pressure splitting;

a condensation reaction that follows a hydrolysis reaction;

a hydroprocessing reaction;

a hydroprocessing reaction and a deoxygenation reaction or a condensation reaction prior to or simultaneous with the hydroprocessing reaction;

a gas removal reaction;

a deoxygenation reaction selected from the group consisting of a hydrogenolysis reaction, hydrogenation, a consecutive hydrogenation-hydrogenolysis reaction, a consecutive hydrogenolysis-hydrogenation reaction, and a combined hydrogenation-hydrogenolysis reaction;

a condensation reaction following a deoxygenation reaction;

an esterification reaction;

an interestification reaction;

a transesterification reaction;

a hydroxylation reaction; and

a condensation reaction following a hydroxylation reaction; and

b. isolating the product of said reaction from the other components.

65. The method of claim 64, wherein the oil-based product is selected from the group consisting of soap, fuel, dielectric fluid, hydraulic fluid, plasticizer, lubricant, heat transfer fluid, and metal working fluid.

66. The method of claim 65, wherein the oil-based product is a fuel product selected from the group consisting of:

a. biodiesel

b. renewable diesel; and

c. jet fuel.

67. The method of claim 66, wherein the fuel product is biodiesel with one or more of the following attributes:

i. less than 0.3 mcg/g total carotenoids;

ii. less than 0.005 mcg/g lycopene;

iii. less than 0.005 mcg/g beta carotene;

iv. less than 0.3 mcg/g chlorophyll A;

v. less than 175 mcg/g gamma tocopherol;

vi. less than 0.25% brassicasterol, campesterol, stignasterol, or beta-sitosterol;

vii. less than 350 mcg/g total tocotrienols;

viii. less than 0.05 mcg/g lutein;

or

ix. less than 275 mcg/g tocopherols.

68. The method of claim 66, wherein the fuel product is renewable diesel that has a T10-T90 of at least 20°

C., 40°

C. or 60°

C.

69. The method of claim 66, wherein the fuel product is jet fuel that meets HRJ-5 and/or ASTM specification D1655.

23. An oleaginous microbial triglyceride oil composition, wherein the fatty acid profile of the triglyceride oil is selected from the group consisting of at least about 1% C8:

0, at least about 1% C10;

0, at least about 1% C12;

0, at least about 2% C14;

0, at least about 30% C16;

0, at least about 5% C18;

0, at least about 60% C18;

1, less than about 7% C18;

2, and at least about 35% saturated fatty acids.

24. The oleaginous microbial triglyceride oil composition of claim 23, produced by cultivating a population of recombinant oleaginous microbial cells in a culture medium, wherein said oleaginous microbial cells comprise an exogenous gene, and wherein optionally, an endogenous desaturase of said oleaginous microbial cells has been inactivated or mutated to have less enzymatic activity.

25. The oleaginous microbial triglyceride oil composition of claim 24, wherein the fatty acid profile of the triglyceride oil is similar to the triglyceride oil profile of a naturally occurring oil.

26. The oleaginous microbial triglyceride oil composition of claim 25, wherein the naturally occurring oil is selected from the group consisting of cocoa butter, coconut oil, palm oil, palm kernel oil, shea butter, beef tallow and lard.

27. The oleaginous microbial triglyceride oil composition of claim 24, wherein the fatty acid profile of the triglyceride oil comprises a profile wherein, the total combined amount of C10:

0, C12;

0 and C14;

0 is at least about 50%, the total combined amount of C16;

0, C18;

0 and C18;

1 is at least about 60%, the total combined amount of C18;

0, C18;

1 and C18;

2 is at least about 60%, the total combined amount of C14;

0, C16;

0, C18;

0 and C18;

1 is at least about 60%, the total combined amounts of C8;

0 and C10;

0 is less than about 50%.

28. The oleaginous microbial triglyceride oil composition of claim 24, wherein the fatty acid profile of the triglyceride oil comprises a ratio of fatty acids selected from the group consisting of C8:

0 to C10;

0 ratio of at least about 5 to 1, C10;

0 to C12;

0 ratio of at least about 6 to 1, C12;

0 to C14;

0 ratio of at least about 5 to 1, C14;

0 to C12;

0 ratio of at least about 7;

1, C14;

0 to C16;

0 ratio of at least about 1 to 2.

29. The oleaginous microbial triglyceride oil composition of claim 24 produced by cultivating a population of recombinant oleaginous microbial cells in a culture medium, wherein said endogenous desaturase is selected from the group consisting of stearoyl ACP desaturase and delta 12 fatty acid desaturase.

30. The oleaginous microbial triglyceride oil composition of claim 29 produced by cultivating a population of recombinant oleaginous microbial cells in a culture medium, produced by cultivating a population of recombinant oleaginous microbial cells in a culture medium, wherein said oleaginous microbial cells comprise an exogenous gene encoding an acyl-ACP thioesterase.

31. The oleaginous microbial triglyceride oil composition of claim 30 produced by cultivating a population of recombinant oleaginous microbial cells in a culture medium, wherein said oleaginous microbial cells comprise an exogenous gene that encodes an acyl-ACP thioesterase selected from the group consisting of Umbellularia californica fatty acyl-ACP thioesterase (GenBank #AAC49001);

Cinnamomum camphora fatty acyl-ACP thioesterase (GenBank #Q39473);

Umbellularia californica fatty acyl-ACP thioesterase (GenBank #Q41635);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank #AAB71729);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank #AAB71730);

Elaeis guineensis fatty acyl-ACP thioesterase (GenBank #ABD83939);

Elaeis guineensis fatty acyl-ACP thioesterase (GenBank #AAD42220);

Populus tomentosa fatty acyl-ACP thioesterase (GenBank #ABC47311);

Arabidopsis thaliana fatty acyl-ACP thioesterase (GenBank #NP_—

172327);

Arabidopsis thaliana fatty acyl-ACP thioesterase (GenBank #CAA85387);

Arabidopsis thaliana fatty acyl-ACP thioesterase (GenBank #CAA85388);

Gossypium hirsutum fatty acyl-ACP thioesterase (GenBank #Q9SQI3);

Cuphea lanceolata fatty acyl-ACP thioesterase (GenBank #CAA54060);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #AAC72882);

Cuphea calophylla subsp. mesostemon fatty acyl-ACP thioesterase (GenBank #ABB71581);

Cuphea lanceolata fatty acyl-ACP thioesterase (GenBank #CAC19933);

Elaeis guineensis fatty acyl-ACP thioesterase (GenBank #AAL15645);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #Q39513);

Gossypium hirsutum fatty acyl-ACP thioesterase (GenBank #AAD01982);

Vitis vinifera fatty acyl-ACP thioesterase (GenBank #CAN81819);

Garcinia mangostana fatty acyl-ACP thioesterase (GenBank #AAB51525);

Brassica juncea fatty acyl-ACP thioesterase (GenBank #ABI18986);

Madhuca longifolia fatty acyl-ACP thioesterase (GenBank #AAX51637);

Brassica napus fatty acyl-ACP thioesterase (GenBank #ABH11710);

Oryza sativa (indica cultivar-group) fatty acyl-ACP thioesterase (GenBank #EAY86877);

Oryza sativa (japonica cultivar-group) fatty acyl-ACP thioesterase (GenBank #NP_—

001068400);

Oryza sativa (indica cultivar-group) fatty acyl-ACP thioesterase (GenBank #EAY99617);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #AAC49269);

Ulmus Americana fatty acyl-ACP thioesterase (GenBank #AAB71731);

Cuphea lanceolata fatty acyl-ACP thioesterase (GenBank #CAB60830);

Cuphea palustris fatty acyl-ACP thioesterase (GenBank #AAC49180);

Iris germanica fatty acyl-ACP thioesterase (GenBank #AAG43858);

Iris germanica fatty acyl-ACP thioesterase (GenBank #AAG43858.1);

Cuphea palustris fatty acyl-ACP thioesterase (GenBank #AAC49179);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank# AAB71729);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank# AAB717291.1);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #U39834);

Umbelluaria californica fatty acyl-ACP thioesterase (GenBank # M94159);

Cinnamomum camphora fatty acyl-ACP thioesterase (GenBank #U31813);

Cuphea wrightii fatty acyl-ACOP thioesterase (GenBank #U56103); and

Ricinus communis fatty acyl-ACP thioesterase (GenBank #ABS30422).

32. The oleaginous microbial triglyceride oil composition of claim 31 produced by cultivating a population of recombinant oleaginous microbial cells in a culture medium, wherein said oleaginous microbial cells further comprise a gene encoding a sucrose invertase.

33. The oleaginous microbial triglyceride oil composition of claim 32 produced by cultivating a population of recombinant oleaginous microbial cells in a culture medium, wherein said oleaginous microbial cells is a cell of a microalgal genus or species selected from Achnanthes orientalis, Agmenellum, Amphiprora hyaline, Amphora coffeiformis, Amphora coffeiformis linea, Amphora coffeiformis punctata, Amphora coffeiformis taylori, Amphora coffeiformis tenuis, Amphora delicatissima, Amphora delicatissima capitata, Amphora sp., Anabaena, Ankistrodesmus, Ankistrodesmus falcatus, Boekelovia hooglandii, Borodinella sp., Botryococcus braunii, Botryococcus sudeticus, Carteria, Chaetoceros gracilis, Chaetoceros muelleri, Chaetoceros muelleri subsalsum, Chaetoceros sp., Chlorella anitrata, Chlorella Antarctica, Chlorella aureoviridis, Chlorella candida, Chlorella capsulate, Chlorella desiccate, Chlorella ellipsoidea, Chlorella emersonii, Chlorella fusca, Chlorella fusca var. vacuolate, Chlorella glucotropha, Chlorella infusionum, Chlorella infusionum var. actophila, Chlorella infusionum var. auxenophila, Chlorella kessleri, Chlorella lobophora (strain SAG 37.88), Chlorella luteoviridis, Chlorella luteoviridis var. aureoviridis, Chlorella luteoviridis var. lutescens, Chlorella miniata, Chlorella minutissima, Chlorella mutabilis, Chlorella nocturna, Chlorella parva, Chlorella photophila, Chlorella pringsheimii, Chlorella protothecoides (including any of UTEX strains 1806, 411, 264, 256, 255, 250, 249, 31, 29, 25, and CCAP strains 211/17 and 211/8d), Chlorella protothecoides var. acidicola, Chlorella regularis, Chlorella regularis var. minima, Chlorella regularis var. umbricata, Chlorella reisiglii, Chlorella saccharophila, Chlorella saccharophila var. ellipsoidea, Chlorella salina, Chlorella simplex, Chlorella sorokiniana, Chlorella sp., Chlorella sphaerica, Chlorella stigmatophora, Chlorella vanniellii, Chlorella vulgaris, Chlorella vulgaris, Chlorella vulgaris f. tertia, Chlorella vulgaris var. autotrophica, Chlorella vulgaris var. viridis, Chlorella vulgaris var. vulgaris, Chlorella vulgaris var. vulgaris f. tertia, Chlorella vulgaris var. vulgaris f. viridis, Chlorella xanthella, Chlorella zofingiensis, Chlorella trebouxioides, Chlorella vulgaris, Chlorococcum infusionum, Chlorococcum sp., Chlorogonium, Chroomonas sp., Chrysosphaera sp., Cricosphaera sp., Cryptomonas sp., Cyclotella cryptica, Cyclotella meneghiniana, Cyclotella sp., Dunaliella sp., Dunaliella bardawil, Dunaliella bioculata, Dunaliella granulate, Dunaliella maritime, Dunaliella minuta, Dunaliella parva, Dunaliella peircei, Dunaliella primolecta, Dunaliella salina, Dunaliella terricola, Dunaliella tertiolecta, Dunaliella viridis, Dunaliella tertiolecta, Eremosphaera viridis, Eremosphaera sp., Ellipsoidon sp., Euglena, Franceia sp., Fragilaria crotonensis, Fragilaria sp., Gleocapsa sp., Gloeothamnion sp., Hymenomonas sp., Isochrysis aff. galbana, Isochrysis galbana, Lepocinclis, Micractinium, Micractinium (UTEX LB 2614), Monoraphidium minutum, Monoraphidium sp., Nannochloris sp., Nannochloropsis salina, Nannochloropsis sp., Navicula acceptata, Navicula biskanterae, Navicula pseudotenelloides, Navicula pelliculosa, Navicula saprophila, Navicula sp., Nephrochloris sp., Nephroselmis sp., Nitschia communis, Nitzschia alexandrina, Nitzschia communis, Nitzschia dissipata, Nitzschia frustulum, Nitzschia hantzschiana, Nitzschia inconspicua, Nitzschia intermedia, Nitzschia microcephala, Nitzschia pusilla, Nitzschia pusilla elliptica, Nitzschia pusilla monoensis, Nitzschia quadrangular, Nitzschia sp., Ochromonas sp., Oocystis parva, Oocystis pusilla, Oocystis sp., Oscillatoria limnetica, Oscillatoria sp., Oscillatoria subbrevis, Pascheria acidophila, Pavlova sp., Phagus, Phormidium, Platymonas sp., Pleurochrysis carterae, Pleurochrysis dentate, Pleurochrysis sp., Prototheca wickerhamii, Prototheca stagnora, Prototheca portoricensis, Prototheca moriformis, Prototheca zopfii, Pyramimonas sp., Pyrobotrys, Sarcinoid chrysophyte, Scenedesmus armatus, Spirogyra, Spirulina platensis, Stichococcus sp., Synechococcus sp., Tetraedron, Tetraselmis sp., Tetraselmis suecica, Thalassiosira weissflogii, and Viridiella fridericiana.

34. The oleaginous microbial triglyceride oil composition of claim 24 produced by cultivating a population of recombinant oleaginous microbial cells in a culture medium, wherein said oleaginous microbial cell is a cell of the genus Prototheca.

35. The oleaginous microbial triglyceride oil composition of claim 34 produced by cultivating a population of recombinant oleaginous microbial cells in a culture medium, wherein said oleaginous microbial cell is a cell of the genus Prototheca moriformis.

36. The oleaginous microbial triglyceride oil of claim 24, wherein said oleaginous microbial cell is an oleaginous yeast cell.

37. The oleaginous microbial triglyceride oil of claim 24, wherein said oleaginous microbial cell is an oleaginous bacterial cell.

38. The oleaginous microbial triglyceride oil composition of claim 26, wherein said naturally occurring oil is coconut oil.

39. The oleaginous microbial triglyceride oil composition of claim 26, wherein said naturally occurring oil is palm oil and wherein said exogenous gene comprises a Elaeis guiniensis thioesterase gene, a Cuphea hookeriana thioesterase gene, a combination of a Cuphea hookeriana KAS IV gene and a Cuphea wrightii FATB2 gene, or a construct designed to disrupt an endogenous KAS II gene.

40. The oleaginous microbial triglyceride oil composition of claim 26, wherein said naturally occurring oil is palm kernel oil and wherein said exogenous gene comprises a combination of a Cuphea wrightii FATB2 gene and a construct designed to disrupt an endogenous SAD2B gene.

41. The oleaginous microbial triglyceride oil composition of claim 26, wherein said naturally occurring oil is shea butter.

42. The oleaginous microbial triglyceride oil composition of claim 26, wherein said naturally occurring oil is beef tallow.

43. The oleaginous microbial triglyceride oil composition of claim 26, wherein said naturally occurring oil is lard and wherein said exogenous gene comprises a combination of U. californica thioesterase gene and a construct designed to disrupt an endogenous SAD2B gene, a combination of a Garcinia mangostana thioesterase gene and a construct designed to disrupt an endogenous SAD2B gene, a Brassica napus thioesterase gene, or a Cuphea hookeriana thioesterase gene.

44. The oleaginous microbial triglyceride oil composition of claim 23, wherein the triglyceride oil composition further comprises an attribute selected from the group consisting of:

i. less than 0.3 mcg/g total carotenoids;

ii. less than 0.005 mcg/g lycopene;

iii. less than 0.005 mcg/g beta carotene;

iv. less than 0.3 mcg/g chlorophyll A;

v. less than 175 mcg/g gamma tocopherol;

vi. less than 0.25% brassicasterol, campesterol, stignasterol, or beta-sitosterol;

vii. less than 350 mcg/g total tocotrienols;

viii. less than 0.05 mcg/g lutein;

or

ix. less than 275 mcg/g tocopherols.

45. A method of producing an oleaginous microbial triglyceride oil composition, said triglyceride oil composition having a fatty acid profile selected from the group consisting of at least about 1% C8:

0, at least about 1% C10;

0, at least about 1% C12;

0, at least about 2% C14;

0, at least about 30% C16;

0, at least about 5% C18;

0, at least about 60% C18;

1, less than about 7% C18;

2, and at least about 35% saturated fatty acids, the method comprising the steps of;

a. cultivating a population of oleaginous microbial cells in a culture medium until at least 10% of the dry cell weight of the oleaginous microbial cells is triglyceride oil; and

b. isolating the triglyceride oil composition from the oleaginous microbial cells.

46. The method of claim 45, wherein said oleaginous microbial cell comprises an exogenous gene, and wherein optionally, an endogenous desaturase of said oleaginous microbial cell has been inactivated or mutated to have less enzymatic activity.

47. The method of claim 46, wherein the fatty acid profile of the triglyceride oil is similar to the triglyceride oil profile of a naturally occurring oil.

48. The method of claim 47, wherein the naturally occurring oil is selected from the group consisting of cocoa butter, coconut oil, palm oil, palm kernel oil, shea butter, beef tallow and lard.

49. The method of claim 46, wherein the fatty acid profile of the triglyceride oil comprises a profile wherein, the total combined amount of C10:

0, C12;

0 and C14;

0 is at least about 50%, the total combined amount of C16;

0, C18;

0 and C18;

1 is at least about 60%, the total combined amount of C18;

0, C18;

1 and C18;

2 is at least about 60%, the total combined amount of C14;

0, C16;

0, C18;

0 and C18;

1 is at least about 60%, the total combined amounts of C8;

0 and C10;

0 is less than about 50%.

50. The method of claim 45, wherein the fatty acid profile of the triglyceride oil comprises a ratio of fatty acids selected from the group consisting of C8:

0 to C10;

0 ratio of at least about 5 to 1, C10;

0 to C12;

0 ratio of at least about 6 to 1, C12;

0 to C14;

0 ratio of at least about 5 to 1, C14;

0 to C12;

0 ratio of at least about 7;

1, and C14;

0 to C16;

0 ratio of at least about 1 to 2.

51. The method of claim 46, said endogenous desaturase is selected from the group consisting of stearoyl ACP desaturase and delta 12 fatty acid desaturase.

52. The method of claim 46, wherein said exogenous gene is selected from the group consisting of a gene encoding an acyl-ACP thioesterase.

53. The method of claim 52, wherein said exogenous gene encodes an acyl-ACP thioesterase selected from the group consisting of Umbellularia californica fatty acyl-ACP thioesterase (GenBank #AAC49001);

Cinnamomum camphora fatty acyl-ACP thioesterase (GenBank #Q39473);

Umbellularia californica fatty acyl-ACP thioesterase (GenBank #Q41635);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank #AAB71729);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank #AAB71730);

Elaeis guineensis fatty acyl-ACP thioesterase (GenBank #ABD83939);

Elaeis guineensis fatty acyl-ACP thioesterase (GenBank #AAD42220);

Populus tomentosa fatty acyl-ACP thioesterase (GenBank #ABC47311);

Arabidopsis thaliana fatty acyl-ACP thioesterase (GenBank #NP_—

172327);

Arabidopsis thaliana fatty acyl-ACP thioesterase (GenBank #CAA85387);

Arabidopsis thaliana fatty acyl-ACP thioesterase (GenBank #CAA85388);

Gossypium hirsutum fatty acyl-ACP thioesterase (GenBank #Q9SQI3);

Cuphea lanceolata fatty acyl-ACP thioesterase (GenBank #CAA54060);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #AAC72882);

Cuphea calophylla subsp. mesostemon fatty acyl-ACP thioesterase (GenBank #ABB71581);

Cuphea lanceolata fatty acyl-ACP thioesterase (GenBank #CAC19933);

Elaeis guineensis fatty acyl-ACP thioesterase (GenBank #AAL15645);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #Q39513);

Gossypium hirsutum fatty acyl-ACP thioesterase (GenBank #AAD01982);

Vitis vinifera fatty acyl-ACP thioesterase (GenBank #CAN81819);

Garcinia mangostana fatty acyl-ACP thioesterase (GenBank #AAB51525);

Brassica juncea fatty acyl-ACP thioesterase (GenBank #ABI18986);

Madhuca longifolia fatty acyl-ACP thioesterase (GenBank #AAX51637);

Brassica napus fatty acyl-ACP thioesterase (GenBank #ABH11710);

Oryza sativa (indica cultivar-group) fatty acyl-ACP thioesterase (GenBank #EAY86877);

Oryza sativa (japonica cultivar-group) fatty acyl-ACP thioesterase (GenBank #NP_—

001068400);

Oryza sativa (indica cultivar-group) fatty acyl-ACP thioesterase (GenBank #EAY99617);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #AAC49269);

Ulmus Americana fatty acyl-ACP thioesterase (GenBank #AAB71731);

Cuphea lanceolata fatty acyl-ACP thioesterase (GenBank #CAB60830);

Cuphea palustris fatty acyl-ACP thioesterase (GenBank #AAC49180);

Iris germanica fatty acyl-ACP thioesterase (GenBank #AAG43858);

Iris germanica fatty acyl-ACP thioesterase (GenBank #AAG43858.1);

Cuphea palustris fatty acyl-ACP thioesterase (GenBank #AAC49179);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank# AAB71729);

Myristica fragrans fatty acyl-ACP thioesterase (GenBank# AAB717291.1);

Cuphea hookeriana fatty acyl-ACP thioesterase (GenBank #U39834);

Umbelluaria californica fatty acyl-ACP thioesterase (GenBank # M94159);

Cinnamomum camphora fatty acyl-ACP thioesterase (GenBank #U31813);

Cuphea wrightii fatty acyl-ACOP thioesterase (GenBank #U56103); and

Ricinus communis fatty acyl-ACP thioesterase (GenBank #ABS30422).

54. The method of claim 46, wherein said oleaginous microbial cell further comprises a gene encoding a sucrose invertase.

55. The method of claim 46, wherein said oleaginous microbial cell is a cell of a microalgal genus or species selected from Achnanthes orientalis, Agmenellum, Amphiprora hyaline, Amphora coffeiformis, Amphora coffeiformis linea, Amphora coffeiformis punctata, Amphora coffeiformis taylori, Amphora coffeiformis tenuis, Amphora delicatissima, Amphora delicatissima capitata, Amphora sp., Anabaena, Ankistrodesmus, Ankistrodesmus falcatus, Boekelovia hooglandii, Borodinella sp., Botryococcus braunii, Botryococcus sudeticus, Carteria, Chaetoceros gracilis, Chaetoceros muelleri, Chaetoceros muelleri subsalsum, Chaetoceros sp., Chlorella anitrata, Chlorella Antarctica, Chlorella aureoviridis, Chlorella candida, Chlorella capsulate, Chlorella desiccate, Chlorella ellipsoidea, Chlorella emersonii, Chlorella fusca, Chlorella fusca var. vacuolate, Chlorella glucotropha, Chlorella infusionum, Chlorella infusionum var. actophila, Chlorella infusionum var. auxenophila, Chlorella kessleri, Chlorella lobophora (strain SAG 37.88), Chlorella luteoviridis, Chlorella luteoviridis var. aureoviridis, Chlorella luteoviridis var. lutescens, Chlorella miniata, Chlorella minutissima, Chlorella mutabilis, Chlorella nocturna, Chlorella parva, Chlorella photophila, Chlorella pringsheimii, Chlorella protothecoides (including any of UTEX strains 1806, 411, 264, 256, 255, 250, 249, 31, 29, 25, and CCAP strains 211/17 and 211/8d), Chlorella protothecoides var. acidicola, Chlorella regularis, Chlorella regularis var. minima, Chlorella regularis var. umbricata, Chlorella reisiglii, Chlorella saccharophila, Chlorella saccharophila var. ellipsoidea, Chlorella salina, Chlorella simplex, Chlorella sorokiniana, Chlorella sp., Chlorella sphaerica, Chlorella stigmatophora, Chlorella vanniellii, Chlorella vulgaris, Chlorella vulgaris, Chlorella vulgaris f. tertia, Chlorella vulgaris var. autotrophica, Chlorella vulgaris var. viridis, Chlorella vulgaris var. vulgaris, Chlorella vulgaris var. vulgaris f. tertia, Chlorella vulgaris var. vulgaris f. viridis, Chlorella xanthella, Chlorella zofingiensis, Chlorella trebouxioides, Chlorella vulgaris, Chlorococcum infusionum, Chlorococcum sp., Chlorogonium, Chroomonas sp., Chrysosphaera sp., Cricosphaera sp., Cryptomonas sp., Cyclotella cryptica, Cyclotella meneghiniana, Cyclotella sp., Dunaliella sp., Dunaliella bardawil, Dunaliella bioculata, Dunaliella granulate, Dunaliella maritime, Dunaliella minuta, Dunaliella parva, Dunaliella peircei, Dunaliella primolecta, Dunaliella salina, Dunaliella terricola, Dunaliella tertiolecta, Dunaliella viridis, Dunaliella tertiolecta, Eremosphaera viridis, Eremosphaera sp., Ellipsoidon sp., Euglena, Franceia sp., Fragilaria crotonensis, Fragilaria sp., Gleocapsa sp., Gloeothamnion sp., Hymenomonas sp., Isochrysis aff. galbana, Isochrysis galbana, Lepocinclis, Micractinium, Micractinium (UTEX LB 2614), Monoraphidium minutum, Monoraphidium sp., Nannochloris sp., Nannochloropsis salina, Nannochloropsis sp., Navicula acceptata, Navicula biskanterae, Navicula pseudotenelloides, Navicula pelliculosa, Navicula saprophila, Navicula sp., Nephrochloris sp., Nephroselmis sp., Nitschia communis, Nitzschia alexandrina, Nitzschia communis, Nitzschia dissipata, Nitzschia frustulum, Nitzschia hantzschiana, Nitzschia inconspicua, Nitzschia intermedia, Nitzschia microcephala, Nitzschia pusilla, Nitzschia pusilla elliptica, Nitzschia pusilla monoensis, Nitzschia quadrangular, Nitzschia sp., Ochromonas sp., Oocystis parva, Oocystis pusilla, Oocystis sp., Oscillatoria limnetica, Oscillatoria sp., Oscillatoria subbrevis, Pascheria acidophila, Pavlova sp., Phagus, Phormidium, Platymonas sp., Pleurochrysis carterae, Pleurochrysis dentate, Pleurochrysis sp., Prototheca wickerhamii, Prototheca stagnora, Prototheca portoricensis, Prototheca moriformis, Prototheca zopfii, Pyramimonas sp., Pyrobotrys, Sarcinoid chrysophyte, Scenedesmus armatus, Spirogyra, Spirulina platensis, Stichococcus sp., Synechococcus sp., Tetraedron, Tetraselmis sp., Tetraselmis suecica, Thalassiosira weissflogii, and Viridiella fridericiana.

56. The method of claim 55, wherein said oleaginous microbial cell is a cell of the genus Prototheca.

57. The method of claim 56, wherein said oleaginous microbial cell is a cell of the genus Prototheca moriformis.

58. The method of claim 48, wherein said naturally occurring oil is coconut oil.

59. The method of claim 48, wherein said naturally occurring oil is palm oil and wherein said exogenous gene comprises a Elaeis guiniensis thioesterase gene, a Cuphea hookeriana thioesterase gene, a combination of a Cuphea hookeriana KAS IV gene and a Cuphea wrightii FATB2 gene, or a construct designed to disrupt an endogenous KAS II gene.

60. The method of claim 48, wherein said naturally occurring oil is palm kernel oil and wherein said exogenous gene comprises a combination of a Cuphea wrightii FATB2 gene and a construct designed to disrupt an endogenous SAD2B gene.

61. The method of claim 48, wherein said naturally occurring oil is shea butter.

62. The method of claim 48, wherein said naturally occurring oil is beef tallow.

63. The method of claim 48, wherein said naturally occurring oil is lard and wherein said exogenous gene comprises a combination of U. californica thioesterase gene and a construct designed to disrupt an endogenous SAD2B gene, a combination of a Garcinia mangostana thioesterase gene and a construct designed to disrupt an endogenous SAD2B gene, a Brassica napus thioesterase gene, or a Cuphea hookeriana thioesterase gene.