Microorganisms and methods for production of specific length fatty alcohols and related compounds

a technology of fatty alcohol and microorganisms, applied in the direction of acyltransferases, dna/rna fragmentation, enzymes, etc., can solve the problems of not producing high yields of fatty alcohol, not commercially viable to produce fatty alcohol directly, and not producing fatty alcohol. achieve the effect of increasing the ratio of nad(p)h to nad(p)

Inactive Publication Date: 2015-10-01
GENOMATICA INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0014]In some embodiments, the non-naturally occurring microbial organism of the invention is Crabtree positive and is in culture medium comprising excess glucose. In such conditions, as described herein, the microbial organism can result in increasing the ratio of NAD(P)H to NAD(P) present in the cytosol of the microbial organism.

Problems solved by technology

Unfortunately, it is not commercially viable to produce fatty alcohols directly from the oxidation of petroleum-based linear hydrocarbons (n-paraffins).
This impracticality is because the oxidation of n-paraffins produces primarily secondary alcohols, tertiary alcohols or ketones, or a mixture of these compounds, but does not produce high yields of fatty alcohols.
Additionally, currently known methods for producing fatty alcohols suffer from the disadvantage that they are restricted to feedstock which is relatively expensive, notably ethylene, which is produced via the thermal cracking of petroleum.
In addition, current methods require several steps, and several catalyst types.

Method used

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  • Microorganisms and methods for production of specific length fatty alcohols and related compounds
  • Microorganisms and methods for production of specific length fatty alcohols and related compounds
  • Microorganisms and methods for production of specific length fatty alcohols and related compounds

Examples

Experimental program
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Effect test

example i

Production of Fatty Alcohols and Fatty Aldehydes by MI-FAE Cycle, MD-FAE Cycle and Acyl-CoA Termination Pathways

[0292]Encoding nucleic acids and species that can be used as sources for conferring fatty alcohol and fatty aldehyde production capability onto a host microbial organism are exemplified further below.

Multienzyme Complexes

[0293]In one exemplary embodiment, the genes fadA and fadB encode a multienzyme complex that exhibits three constituent activities of the malonyl-CoA independent FAS pathway, namely, ketoacyl-CoA thiolase, 3-hydroxyacyl-CoA dehydrogenase, and enoyl-CoA hydratase activities (Nakahigashi, K. and H. Inokuchi, Nucleic Acids Research 18:4937 (1990); Yang et al., Journal of Bacteriology 173:7405-7406 (1991); Yang et al, Journal of Biological Chemistry 265:10424-10429 (1990); Yang et al., Biochemistry 30:6788-6795 (1990)). The fadI and fadJ genes encode similar activities which can substitute for the above malonyl-CoA independent FAS conferring genes fadA and fad...

example ii

Pathways for Producing Cytosolic Acetyl-CoA from Cytosolic Pyruvate

[0331]The following example describes exemplary pathways for the conversion of cytosolic pyruvate and threonine to cytosolic acetyl-CoA, as shown in FIG. 2.

[0332]Pathways for the conversion of cytosolic pyruvate and threonine to cytosolic acetyl-CoA could enable deployment of a cytosolic fatty alcohol, fatty aldehyde or fatty acid production pathway that originates from acetyl-CoA. Several pathways for converting cytosolic pyruvate to cytosolic acetyl-CoA are shown in FIG. 2. Direct conversion of pyruvate to acetyl-CoA can be catalyzed by pyruvate dehydrogenase, pyruvate formate lyase, pyruvate:NAD(P) oxidoreductase or pyruvate:ferredoxin oxidoreductase. If a pyruvate formate lyase is utilized, the formate byproduct can be further converted to CO2 by formate dehydrogenase or formate hydrogen lyase.

[0333]Indirect conversion of pyruvate to acetyl-CoA can proceed through several alternate routes. Pyruvate can be convert...

example iii

Pathways for Producing Acetyl-CoA from PEP and Pyruvate

[0352]Pathways for the conversion of cytosolic phosphoenolpyruvate (PEP) and pyruvate to cytosolic acetyl-CoA can also enable deployment of a cytosolic fatty alcohol, fatty aldehyde or fatty acid production pathway from acetyl-CoA. FIG. 3 shows numerous pathways for converting PEP and pyruvate to acetyl-CoA.

[0353]The conversion of PEP to oxaloacetate is catalyzed in one, two or three enzymatic steps. Oxaloacetate is further converted to acetyl-CoA via malonate semialdehyde or malonyl-CoA intermediates. In one pathway, PEP carboxylase or PEP carboxykinase converts PEP to oxaloacetate (step A); oxaloacetate decarboxylase converts the oxaloacetate to malonate (step B); and malonate semialdehyde dehydrogenase (acetylating) converts the malonate semialdehyde to acetyl-CoA (step C). In another pathway pyruvate kinase or PEP phosphatase converts PEP to pyruvate (step N); pyruvate carboxylase converts the pyruvate to (step H); oxaloacet...

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Abstract

The invention provides non-naturally occurring microbial organisms containing a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms selectively produce a fatty alcohol, fatty aldehyde or fatty acid of a specified length. Also provided are non-naturally occurring microbial organisms having a fatty alcohol, fatty aldehyde or fatty acid pathway, wherein the microbial organisms further include an acetyl-CoA pathway. In some aspects, the microbial organisms of the invention have select gene disruptions or enzyme attenuations that increase production of fatty alcohols, fatty aldehydes or fatty acids. The invention additionally provides methods of using the above microbial organisms to produce a fatty alcohol, a fatty aldehyde or a fatty acid.

Description

[0001]This application claims the benefit of priority of U.S. Provisional application Ser. No. 61 / 714,144, filed Oct. 15, 2012, the entire contents of which is incorporated herein by reference.BACKGROUND OF THE INVENTION[0002]The present invention relates generally to biosynthetic processes, and more specifically to organisms having specific length fatty alcohol, fatty aldehyde or fatty acid biosynthetic capacity.[0003]Primary alcohols are a product class of compounds having a variety of industrial applications which include a variety of biofuels and specialty chemicals. Primary alcohols also can be used to make a large number of additional industrial products including polymers and surfactants. For example, higher primary alcohols, also known as fatty alcohols (C4-C24) and their ethoxylates are used as surfactants in many consumer detergents, cleaning products and personal care products worldwide such as laundry powders and liquids, dishwashing liquid and hard surface cleaners. The...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12P7/64C12N9/04C12N9/88C07C31/20C12N9/00C12N9/12C12N9/16C12N9/10C12N9/02
CPCC12P7/64C12Y604/01001C12N9/0006C12N9/88C12N9/001C12N9/0008C12N9/93C12N9/1217C12N9/1294C12N9/13C12N9/16C12N9/1025C07C31/207C12Y101/01035C12Y101/01037C12Y101/05004C12Y101/05006C12Y101/99033C12Y102/01C12Y102/01002C12Y102/01003C12Y102/0101C12Y102/01015C12Y102/01018C12Y102/01042C12Y102/0105C12Y102/01076C12Y102/01027C12Y102/05001C12Y103/01044C12Y103/99012C12Y203/01008C12Y203/01016C12Y203/01054C12Y203/01174C12Y203/03008C12Y207/02001C12Y207/02007C12Y207/09002C12Y208/03003C12Y301/03001C12Y301/03002C12Y301/0302C12Y301/0306C12Y401/01C12Y401/01001C12Y401/01003C12Y401/01007C12Y401/01009C12Y401/01031C12Y401/01032C12Y401/01049C12Y401/01074C12Y401/01082C12Y401/03006C12Y401/03034C12Y401/02005C12Y402/01017C12Y602/01001C12Y602/01013C12Y602/01018C12N9/1029C07C29/00C07C31/02C07C33/00C07C47/263C07C53/00C07C57/00C12N1/00C12P7/04C12P7/40C12N15/52C12P7/6409C12P7/24Y02E50/10
Inventor OSTERHOUT, ROBIN E.BURGARD, ANTHONY P.
Owner GENOMATICA INC
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