Production of fatty acid derivatives

a technology of fatty acid derivatives and derivatives, which is applied in the direction of acyltransferases, microorganisms, enzymes, etc., can solve the problems of high cost of petroleum products development, high cost, and high cost of petroleum exploration, and achieve low impurities and/or undesirable contaminants, and clean emission profiles

Inactive Publication Date: 2011-07-07
LS9 INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0063]In a further aspect, the invention features a method of producing a biologically-derived diesel fuel of commercial quality according to commercial standards (e.g., ASTM or ANP). In some embodiments, the method comprises fermenting carbohydrates using a genetically modified microorganism described herein. The process provides a direct route for producing fatty esters, for example, fatty acid esters such as fatty acid methyl esters or fatty acid ethyl esters, without the need of producing oils, which are later chemically transesterified with the concomitant production of large quantities of glycerol. The fuel composition thus produced can be utilized as a diesel fuel alone, or be blended with petroleum diesel according to customary proportions, resulting in clean emission profiles and low amounts of impurities and / or undesirable contaminants.

Problems solved by technology

Petroleum is a valuable resource, but petroleum products are developed at considerable costs, both financial and environmental.
First, sources of petroleum must be discovered.
Petroleum exploration is an expensive and risky venture.
In addition to the economic cost, petroleum exploration carries a high environmental cost.
For example, offshore exploration disturbs the surrounding marine environments.
After a productive well is discovered, the petroleum must be extracted from the Earth at great expense.
Petroleum extraction also carries an environmental cost.
For example, petroleum extraction can result in large seepages of petroleum rising to the surface.
Offshore drilling involves dredging the seabed which disrupts or destroys the surrounding marine environment.
In addition to the shipping costs, there is also the environmental risk of devastating oil spills.
Obtaining these specialty chemicals from crude petroleum requires a significant financial investment as well as a great deal of energy.
It is also an inefficient process because frequently the long chain hydrocarbons in crude petroleum are cracked to produce smaller monomers.
In addition to the problems with exploring, extracting, transporting, and refining petroleum, petroleum is a limited and dwindling resource.
As the world's demand for fuel increases, the emission of greenhouse gases and other forms of air pollution also increases.
Hence, in addition to damaging the environment locally (e.g., oil spills, dredging of marine environments, etc.), burning petroleum also damages the environment globally.
Industrial-scale biodiesel production is thus geographically and seasonally restricted to areas where vegetable oil feedstocks are produced.
However, glycerin is an undesirable byproduct of the transesterification process.
This increases costs and the amount of energy required for fatty ester production and, ultimately, biodiesel production as well.
Furthermore, vegetable oil feedstocks are inefficient sources of energy because they require extensive acreage for cultivation.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Production of E. coli MG1655 ΔfadE

[0240]This example describes the construction of a genetically engineered microorganism wherein the expression of a fatty acid degradation enzyme is attenuated.

[0241]The fadE gene of E. coli MG1655 was deleted using the Lambda Red (also known as the Red-Driven Integration) system described in Datsenko et al., Proc. Natl. Acad. Sci. USA 97: 6640-6645 (2000), with the following modifications.

[0242]Two primers were used to create the deletion:

Del-fadE-F:(SEQ ID NO: 1)5′-AAAAACAGCAACAATGTGAGCTTTGTTGTAATTATATTGTAAACATATTGATTCCGGGGATCCGTCGACC-3′Del-fadE-R:(SEQ ID NO: 2)5′-AAACGGAGCCTTTCGGCTCCGTTATTCATTTACGCGGCTTCAACTTTCCTGTAGGCTGGAGCTGCTTC-3′

[0243]The Del-fadE-F and Del-fadE-R primers were used to amplify the Kanamycin resistance (KmR) cassette from plasmid pKD13 (as described in Datsenko et al., supra) by PCR. The PCR product was then used to transform electrocompetent E. coli MG1655 cells containing pKD46 (described in Datsenko et al., supra). These cel...

example 2

Production of E. coli MG1655 ΔfadE ΔfhuA

[0245]This example describes the construction of a genetically engineered microorganism in which the expression of a fatty acid degradation enzyme and an outer membrane protein receptor are attenuated.

[0246]The fhuA (also known as tonA) gene of E. coli MG1655, which encodes a ferrichrome outer membrane transporter (GenBank Accession No. NP—414692), was deleted from strain E. coli MG1655 D1 of Example 1 using the Lambda Red system described in Datsenko et al., supra, but with the following modifications.

[0247]Two primers were used to create the deletion:

Del-fhuA-F:(SEQ ID NO: 5)5′-ATCATTCTCGTTTACGTTATCATTCACTTTACATCAGAGATATACCAATGATTCCGGGGATCCGTCGACC-3′;Del-fhuA-R:(SEQ ID NO: 6)5′-GCACGGAAATCCGTGCCCCAAAAGAGAAATTAGAAACGGAAGGTTGCGGTTGTAGGCTGGAGCTGCTTC-3′

[0248]The Del-fhuA-F and Del-fhuA-R primers were used to amplify the KmR cassette from plasmid pKD13 by PCR. The PCR product obtained was used to transform the electrocompetent E. coli MG1655 D1 c...

example 3

Production of E. coli MG1655 ΔfadE ΔfhuA ΔpflB ΔldhA

[0251]This example describes the construction of a genetically engineered microorganism in which the expression of an acyl-CoA dehydrogenase, an outer membrane protein receptor, a pyruvate formate lyase and a lactate dehydrogenase are attenuated.

[0252]The pflB gene of E. coli MG1655, which encodes a pyruvate formate lyase (GenBank Accession No. AAC73989), was deleted from E. coli MG1655 DV2 (see, Example 2) using the Lambda Red System according to Datsenko et al., supra, but with the following modifications:

[0253]The primers used to create the deletion strain were:

Del-pflB-F:(SEQ ID NO: 33)5′-GCCGCAGCCTGATGGACAAAGCGTTCATTATGGTGCTGCCGGTCGCGATGATTCCGGGGATCCGTCGACC-3′Del-pflB-R: (SEQ ID NO: 34)5′-ATCTTCAACGGTAACTTCTTTACCGCCATGCGTGTCCCAGGTGTCTGTAGGCTGGAGCTGCTTCG-3′

[0254]The Del-pflB-F and Del-pflB-R primers were used to amplify the Kanamycin resistance (KmR) cassette from plasmid pKD13 by PCR. The PCR product was then used to transform...

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Abstract

Methods and compositions for producing fatty acid derivatives, for example, fatty esters, and commercial fuel compositions comprising fatty acid derivatives are described.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims the benefit of U.S. Provisional Application No. 61 / 245,943, filed Sep. 25, 2009, the contents of which are hereby incorporated in their entirety herein.BACKGROUND OF THE INVENTION[0002]Petroleum is a limited, natural resource found in the Earth in liquid, gaseous, or solid forms. Petroleum is primarily composed of hydrocarbons, which are comprised mainly of carbon and hydrogen. It also contains significant amounts of other elements, such as, nitrogen, oxygen, or sulfur, in different forms.[0003]Petroleum is a valuable resource, but petroleum products are developed at considerable costs, both financial and environmental. First, sources of petroleum must be discovered. Petroleum exploration is an expensive and risky venture. The cost of exploring deep water wells can exceed $100 million. In addition to the economic cost, petroleum exploration carries a high environmental cost. For example, offshore exploration distur...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C10L1/19C07C53/126C07C69/003C12P7/64C12N1/00C12N1/21
CPCC10L1/026C10L1/19C11C3/10C12N9/1029C12P7/649Y02E50/13C12N1/36C10L1/04C12N15/63C12N9/001C12Y103/99003C12Y203/01075Y02E50/10
Inventor GAERTNER, ALFRED
Owner LS9 INC
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