Enhanced production of fatty acid derivatives

A technology of fatty acid derivatives and fatty alcohols, applied in the field of increasing the output of fatty acid derivatives, can solve the problems of increasing biodiesel production, increasing the cost of fatty esters and required energy, and vegetable oil raw materials are not efficient energy sources

Active Publication Date: 2010-03-24
GENOMATICA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This ultimately increases biodiesel production, but increases the cost and energy required for fatty ester production
Also, vegetable oil feedstocks are not efficient sources of energy because they require extensive cultivation areas

Method used

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  • Enhanced production of fatty acid derivatives
  • Enhanced production of fatty acid derivatives
  • Enhanced production of fatty acid derivatives

Examples

Experimental program
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Effect test

preparation example Construction

[0151] I. Preparation of Fatty Acid Derivatives and Modifications to Improve Yield

[0152] Production hosts for the production of acyl-CoA and / or fatty acid derivatives can be recombinantly modified to include nucleic acid sequences for overexpressing peptides. For example, production hosts can be modified to increase the production of acyl-CoA, reduce the metabolism of fatty acid derivatives and intermediates in the fatty acid biosynthetic pathway such as acyl-CoA, or reduce feedback inhibition at specific points in the fatty acid biosynthetic pathway. In addition to modifying the genes described herein, other cellular resources can also be diverted to overproduce fatty acids, for example the lactate, succinate and / or acetate pathways can be attenuated and acetyl-CoA carboxylase (acc ). Modifications to the production hosts described herein can be accomplished by genomic alterations, addition of recombinant expression systems, or combinations thereof.

[0153] Figure 2-F...

Embodiment 1

[0348] Embodiment 1. Construction of production host

[0349]An exemplary production host is LS9001. LS9001 was prepared by modifying C41(DE3) purchased from Overexpress.com (Saint Beausine, France) to functionally delete the fadE gene (acyl-CoA dehydrogenase).

[0350] Briefly, primers YafV_NotI and Ivry_O1 were used to amplify about 830 bp upstream of fadE, and primers Lpcaf_o1 and LpcaR_Bam were used to amplify about 960 bp downstream of fadE to prepare a fadE knockout strain of E. coli. Overlap PCR was used to generate a construct with an in-frame deletion of the entire fadE gene. The fadE deletion construct was cloned into the temperature-sensitive plasmid pKOV3, which contains the sacB gene for counter-selection, and chromosomal deletion of fadE was achieved following the method of Link et al. (J. Bact. 179:6228-6237, 1997). The obtained strain cannot degrade fatty acids and acyl-CoA. This knockout strain is referred to herein as ΔfadE.

[0351] Other modifications t...

Embodiment 2

[0358] Example 2. Modification of Production Host

[0359] The following plasmids were constructed for expression of various proteins for the synthesis of fatty acid derivatives. Constructs are prepared using standard molecular biology methods. All cloned genes were placed under the control of an IPTG-inducible promoter (eg, T7, tac or lac promoter).

[0360] The 'tesA gene of Escherichia coli (thioesterase A gene, accession number NP_415027, without leader sequence (Cho and Cronan, The J.of Biol.Chem., 270: 4216-9, 1995, EC: 3.1.1.5, 3.1.2.-) Cloning into NdeI / AvrII digested pETDuet-1 (pETDuet-1 described herein was obtained from Novagen, Madison, WI). FatB-type phytothioesters encoding laurel california, calyx japonica and camphor tree The genes of enzymes (TEs) (accession numbers: UcFatB1=AAA34215, ChFatB2=AAC49269, ChFatB3=AAC72881, CcFatB=AAC49151) were respectively cloned into three different vectors: (i) pETDuet-1 digested by NdeI / AvrII, (ii) XhoI / HindIII digested pB...

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Abstract

Genetically engineered cells and microorganisms are provided that produce products from the fatty acid biosynthetic pathway (fatty acid derivatives), as well as methods of their use. The products areparticularly useful as biofuels.

Description

technical field [0001] The present invention provides genetically engineered cells and microorganisms that produce products from fatty acid biosynthetic pathways (ie, fatty acid derivatives), and methods of their use. Such products are particularly suitable as biofuels. [0002] Cross References to Related Applications [0003] This application claims the benefit of U.S. Provisional Application Serial No. 60 / 908,547, filed March 28, 2007, U.S. Provisional Application Serial No. 60 / 989,798, filed November 21, 2007, and PCT Application PCT / priority of US2007 / 011923, both of which are incorporated herein by reference in their entirety. Background technique [0004] As technology develops, with it comes an increased dependence on fuel sources. Such fuel sources are becoming increasingly limited and difficult to obtain. With fossil fuels being burned at an unprecedented rate, the world's fuel needs may soon outstrip current fuel supplies. [0005] Therefore, efforts are now...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/64C12N9/02C12N9/04C12N9/10C12N9/16C10L1/02C12N1/19C12N1/21
CPCC12P7/6409C12P7/6436Y02E50/13C12N9/93Y02E50/343C12P7/649C12P7/6463C10G2300/1011C12N9/16C12N9/001C12N9/0008C12N9/1025C12N9/0006C12N9/1029C12Y101/01C12Y101/01001C12Y102/0105C12Y103/99003C12Y203/0102C12Y203/01075C12Y301/02C12Y602/01Y02P30/20Y02P20/59C12P7/04Y02E50/10C12N1/20C12N1/18C12N15/52C12P7/64C12P7/6458
Inventor 胡志浩费尔南多·瓦勒
Owner GENOMATICA INC
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