Glycerol feedstock utilization for oil-based fuel manufacturing

• US 8,697,402 B2
• Filed: 08/19/2008
• Issued: 04/15/2014
• Est. Priority Date: 06/01/2007
• Status: Expired due to Fees

• Current Assignee: Corbion Biotech, Inc. (Corbion NV)
• Original Assignee: Solazyme Inc.
• Inventors: Trimbur, Donald E., Im, Chung-Soon, Dillon, Harrison F., Day, Anthony G., Franklin, Scott, Coragliotti, Anna
• Primary Examiner(s): BERTOGLIO, VALARIE E
• Application Number:
  US12/194,389
• Publication Number:
  US 20090148918A1
• Time in Patent Office: 2,065 Days
• Field of Search: 435/134
• US Class Current: 435/134
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C10L 1/026 for compression ignition
• CPC Class Codes: C10L 1/06 for spark ignition
• CPC Class Codes: C10L 2200/0484 Vegetable or animal oils
• CPC Class Codes: C10L 2270/026 for diesel engines, e.g. automobiles, stationary, marine
• CPC Class Codes: C10L 2270/04 for turbines, planes, power generation
• 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 1/12 Unicellular algae; Culture media therefor as new plants A01H13/00
• CPC Class Codes: C12N 1/32 Processes using, or culture media containing, lower alkanols, i.e. C1 to C6
• CPC Class Codes: C12N 15/79 Vectors or expression systems specially adapted for eukaryotic hosts
• CPC Class Codes: C12N 9/0006 acting on CH-OH groups as donors (1.1)
• CPC Class Codes: C12N 9/0008 acting on the aldehyde or oxo group of donors (1.2)
• CPC Class Codes: C12N 9/1205 Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases
• CPC Class Codes: C12N 9/14 Hydrolases (3)
• CPC Class Codes: C12N 9/16 acting on ester bonds (3.1)
• CPC Class Codes: C12N 9/20 Triglyceride splitting, e.g. by means of lipase
• CPC Class Codes: C12N 9/2408 acting on alpha -1,4-glucosidic bonds
• CPC Class Codes: C12N 9/2431 Beta-fructofuranosidase (3.2.1.26), i.e. invertase
• CPC Class Codes: C12N 9/88 Lyases (4.)
• CPC Class Codes: C12P 21/06 produced by the hydrolysis of a peptide bond, e.g. hydrolysate products preparing foodstuffs by protein hydrolysis A23J3/00
• CPC Class Codes: C12P 5/02 : acyclic C12P5/007 takes precedence
• CPC Class Codes: C12P 7/6409 : Fatty acids
• CPC Class Codes: C12P 7/6458 : by transesterification, e.g. interesterification, ester interchange, alcoholysis or acidolysis
• CPC Class Codes: C12P 7/6463 : obtained from glyceride producing microorganisms, e.g. single cell oil
• CPC Class Codes: C12P 7/649 : Biodiesel, i.e. fatty acid alkyl esters
• 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: Y02E 50/30 : Fuel from waste, e.g. synthetic alcohol or diesel
• CPC Class Codes: Y02P 20/52 : using catalysts, e.g. selective catalysts
• CPC Class Codes: Y02T 50/678 : Aviation using fuels of non-fossil origin

Abstract

The invention provides methods of manufacturing biodiesel and other oil-based compounds using glycerol and combinations of glycerol and other feedstocks as an energy source in fermentation of oil-bearing microorganisms. Methods disclosed herein include processes for manufacturing high nutrition edible oils from non-food feedstock materials such as waste products from industrial waste transesterification processes. Also included are methods of increasing oil yields by temporally separating glycerol and other feedstocks during cultivation processes. Also provided herein are oil-bearing microbes containing exogenous oil production genes and methods of cultivating such microbes on glycerol and other feedstocks.

Claims

1. A method of triglyceride manufacturing, the method comprising:

(a) heterotrophically culturing a microalga in a culture medium comprising glycerol supplied to the culture medium at a concentration of at least 500 mM as a source of fixed carbon in the dark supporting production of microbial biomass, wherein the glycerol is a byproduct of a transesterification process and is contaminated with methanol or ethanol from the transesterification process and/or wherein the glycerol is non-acidulated; and

(b) separating the triglyceride from the microbial biomass using simultaneous heat and mechanical lysis.

2. The method of claim 1, wherein the microalga is selected from the group consisting of the microalgalisted in Table 1.

3. The method of claim 2, wherein the microalga is a species of the genus Chlorella.

4. The method of claim 3, wherein the microalga is selected from the group consisting of Chlorella anitrata, Chlorella antarctica, Chlorella aureoviridis, Chlorella candida, Chlorella capsulata, Chlorella desiccata, 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 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, 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. airidis, Chlorella vulgaris var. vulgaris, Chlorella vulgaris var. vulgaris f. tertia, Chlorella vulgaris var. vulgaris f. viridis, Chlorella xanthella, and Chlorella zofingiensis.

5. The method of claim 4, wherein the microalga is selected from the group consisting of Chlorella kessleri UTEX strain #263, Chlorella kessleri UTEX strain #397, Chlorella kessleri UTEX strain #398, Chlorella kessleri UTEX strain #2228, Chlorella sorokiniana UTEX strain #1663, Chlorella sorokiniana UTEX strain #1665, Chlorella sorokiniana UTEX strain #1669, Chlorella sorokiniana UTEX strain #1671, Chlorella sorokiniana UTEX strain #1810, Chlorella saccharophila UTEX strain #2911, Chlorella saccharophila UTEX strain #2469, Chlorella protothecoides UTEX strain #31, Chlorella protothecoides UTEX strain #249, Chlorella protothecoides UTEX strain #250, and Chlorella protothecoides UTEX strain #264.

6. The method of claim 1, wherein the microalga contains at least one exogenous sucrose utilization gene.

7. The method of claim 6, wherein the sucrose utilization gene encodes a sucrose transporter, a sucrose invertase, a hexokinase, a glucokinase, or a fructokinase.

8. The method of claim 1, wherein the microalga contains at least one exogenous gene encoding a lipid pathway enzyme.

9. The method of claim 8, wherein the lipid pathway enzyme is selected from the group consisting of a stearoyl-ACP desaturase, a glycerolipid desaturase, a pyruvate dehydrogenase, an acetyl-CoA carboxylase, a fatty acyl-ACP thioesterase, an acyl carrier protein, and a glycerol-3 phosphate acyltransferase.

10. The method of claim 1, further comprising the step of chemically modifying the separated triglyceride to produce diesel, wherein the diesel is biodiesel or renewable diesel.

11. The method of claim 1, further comprising the step of deoxygenating the lipid.

12. The method of claim 1, further comprising the step of hydrogenating the lipid.

13. The method of claim 1, further comprising the step of hydrocracking the lipid.

14. The method of claim 1, further comprising one or more steps selected from the list consisting of deoxygenating the lipid, hydrogenating the lipid, and hydrocracking the lipid.

15. The method of claim 1, further comprising the step of processing the isolated lipid to produce jet fuel.

16. The method of claim 15, wherein the processing of the isolated lipid to produce jet fuel comprises fluid catalytic cracking.

17. The method of claim 15, wherein the processing of the isolated lipid to produce jet fuel comprises hydrodeoxygenation.

18. The method of claim 1, further comprising the step of transesterifying the triglyceride.

19. The method of claim 1, wherein said culturing step takes place under limiting nitrogen conditions.

20. The method of claim 1, wherein the glycerol is contaminated with residual methanol from the transesterification process.

21. The method of claim 1, wherein the glycerol is non-acidulated.

22. The method of claim 6, wherein the source of fixed carbon further comprises sucrose.