JP2012515795A - Method for producing dodecanedioic acid and its derivatives - Google Patents

Abstract

Methods for purifying dodecanedioic acid derived from biological sources and compositions comprising dodecanedioic acid derived from biological sources are provided. In some embodiments, the method first comprises biologically forming muconic acid from a renewable carbon source, reducing the muconic acid to hexenedioic acid, and then the hexenedioic acid. It includes an acid (typically, delta ⁹ unsaturated fatty acid) unsaturated fatty acids and are reacted in the metathesis reaction, a step of generating dodecene diacid. The dodecenedioic acid is then reduced to dodecanedioic acid. Dodecanedioic acid can be used to form polymers (eg, polyamides). Examples of the polyamide include nylon (for example, nylon 6,12). Nylon 6,12 can be formed by reacting dodecanedioic acid with 1,6-hexamethylenediamine.

Description

(Related application)
This application claims the benefit of US Provisional Application No. 61 / 146,545, filed Jan. 22, 2009, which is hereby incorporated by reference in its entirety.

(Field of Invention)
The present invention generally relates to the production of dodecanedioic acid from renewable feedstocks and their subsequent use (eg, to form polyamides).

(Background of the Invention)
Nylon is a generic name for a family of synthetic thermoplastic polyamides used to make fabrics, musical strings, ropes, screws and gears, to name but a few. . Nylon can also be used as a filler (eg, glass filled variants and molybdenum sulfide filled variants).

  Nylon 6 is the most common commercial grade molded nylon. Many suffixes specify the number of carbon atoms provided by the monomer (diamine first and diacid second). For nylon 6,6, the diamine is typically hexamethylene diamine and the diacid is adipic acid. Each of these monomers provides 6 carbons to the polymer chain.

  Another example of a useful nylon is nylon 6,12, which is a copolymer of a 6 carbon diamine and a 12 carbon dicarboxylic acid. One method for making nylon 6,12 involves forming a polycondensation product of 1,6-hexamethylenediamine and dodecanedioic acid. For the commercial production of such polymeric materials, the starting material is obtained substantially solely from a hydrocarbon source.

  Dodecanedioic acid is therefore a very important compound. It is used in various industrial applications such as plasticizers for polymers, epoxy curing agents, adhesives and powder coatings, engineering plastics, perfume products and pharmaceuticals. In one year, 15,000,000,000 pounds of dodecanedioic acid are synthesized from petrochemical feedstock. Such petrochemical feedstocks are primarily natural resources that are being depleted and the use of such feedstocks has been linked to harmful changes to the environment on a global scale.

  Such feedstock materials useful for the production of nylon are therefore limited in availability and subject to substantial price fluctuations. As a result, there is an increasing interest and need for dodecanedioic acid, as well as alternative methods for producing polyamides that are renewable and more harmless to sustainable environments.

(Summary of the Invention)
An aspect of the present invention is the production of useful commercial products (eg, polyamides) that use starting materials obtained from non-renewable feedstocks (eg, petroleum or other fossil carbon resources). Symmetrically, starting with materials produced by biological processes from renewable feedstocks). Aspects of the present invention more specifically relate to the production of dicarboxylic acids and derivatives thereof from renewable biomass-derived carbon sources. More specifically, some aspects of the present invention relate to the production of dodecanedioic acid and its precursors and derivatives from renewable biomass-derived carbon sources. More specifically, the method of the present invention provides a metathesis step with an olefin compound to produce a biosourced dicarboxylic acid (eg, dodecanedioic acid) from a renewable source of biosource. Is used. The resulting renewable dodecanedioic acid can be separated from other products of the metathesis reaction and from any remaining starting materials. Dodecanedioic acid and its derivatives have utility in the production of polyamides and other polymers.

  In some aspects, the present invention first provides a method for biologically producing muconic acid from a renewable feedstock. In a preferred embodiment, the muconic acid is reduced to single or multiple isomers of hexenedioic acid. The reduction of the muconic acid can be performed using methods known in the art (eg, zinc halide reagents, electrochemical reduction, or selective hydrogenation). The hexenedioic acid can exist as a derivative (eg, ester, amide, or salt).

  In some embodiments, the hexenedioic acid is reacted with an unsaturated fatty acid in a metathesis reaction to produce dodecenedioic acid, which is then reduced to dodecanedioic acid. The above reaction is typically a metathesis catalyst (eg, Grubbs catalyst (benzylidene-bis (tricyclohexylphosphine) dichlororuthenium or benzylidene [1,3-bis (2,4,6-trimethylphenyl) -2-imidazolidini). Use of (Ridene) dichloro (tricyclohexylphosphine) ruthenium).

  Biological formation of muconic acid can be attributed to Escherichia genus, Klebsiella genus, Corynebacterium genus, Brevibacterium genus, Arthrobacter genus, Bacillus genus, Pseudomonas genus, Streptomyces genus, Staphylococcus Or using a yeast of the genus Saccharomyces or Schizosaccharomyces, may comprise the step of forming the muconic acid.

  Muconic acid is reduced to the isomer of hexenedioic acid (eg, 3-hexenedioic acid) using any suitable reagent. Another suitable reagent is a zinc halide reagent (eg, zinc chloride in pyridine).

The hexenedioic acid or derivative thereof is reacted with an unsaturated fatty acid to form an unsaturated dicarboxylic acid or derivative thereof. In a preferred embodiment, the unsaturated fatty acid is first reacted in a self-metathesis reaction to produce Δ ⁹ octadecenedioic acid. Δ ⁹ octadecenedioic acid then reacts with the hexenedioic acid to produce dodecenedioic acid. Preferably, the unsaturated fatty acid is a delta ⁹ unsaturated fatty acids. Examples of the Δ ⁹ unsaturated fatty acid include, but are not limited to, myristoleic acid, palmitoleic acid, elaidic acid, and oleic acid. The unsaturated dicarboxylic acid or derivative thereof produced by the metathesis reaction can then be reduced to a saturated dicarboxylic acid. In a preferred embodiment, the unsaturated dicarboxylic acid formed is dodecenedioic acid, which is then reduced to its saturated analog, dodecanedioic acid. This can be accomplished, for example, by hydrogenating the dodecenedioic acid using a noble metal hydrogenation catalyst.

In another embodiment, the Δ ⁹ unsaturated fatty acid is first converted to octadecenedioic acid, a symmetric Δ ⁹ unsaturated dicarboxylic acid, via a self-metathesis reaction. Then, the symmetry of the delta ^9-octadecene diacid, in cross metathesis reactions, is used together with the symmetry 3-hexene diacid may provide desired dodecene diacid as a single product of the metathesis reaction.

  In some embodiments, the dodecanedioic acid is used to form a polymer (eg, polyamide). Examples of the polyamide include nylon (for example, nylon 6,12). Nylon 6,12 can be formed by reacting 1,6-hexamethylenediamine and dodecanedioic acid.

Particular embodiments of the disclosed invention include, firstly, the genus Escherichia, Klebsiella, Corynebacterium, Brevibacterium, Arthrobacter, Bacillus, Pseudomonas, Streptomyces, Using a yeast of the genus Saccharomyces or Schizosaccharomyces to biologically form muconic acid. The muconic acid is reduced to 3-hexenedioic acid using a zinc halide reagent. The 3-hexene diacid, delta ⁹ unsaturated fatty acids (e.g., myristoleic acid, palmitoleic acid, elaidic acid, oleic acid, or combinations thereof) and, is using a metathesis catalyst is reacted in a metathesis reaction, This produces dodecenedioic acid. The dodecenedioic acid is reduced by hydrogenation to form dodecanedioic acid. The dodecanedioic acid is then used to form a polyamide (eg, nylon 6,12) by reaction with a suitable diamine (eg, 1,6-hexamethylenediamine).

Aspects of the invention relate to compositions comprising unsaturated dicarboxylic acids derived from biological sources or derivatives thereof, and saturated analogs thereof produced from renewable feedstocks derived from biomass. In some embodiments, the composition comprises dodecenedioic acid or a dodecenedioic acid derivative derived from a biological source. In preferred embodiments, disclosed are products derived from biological sources having a carbon isotope distribution or ¹⁴ C / ¹² C ratio characteristic of products synthesized from renewable carbon sources. In some embodiments, the renewable, isolated dodecenedioic acid or dodecenedioic acid derivative thereof is greater than 0, greater than 0.9 × 10 ⁻¹² , or about 1.2 × 10 ^−12. Characterized by a ¹⁴ C / ¹² C ratio of In some embodiments, the dodecenedioic acid derivative is dimethyl dodecenedioic acid. In other embodiments, a composition comprising dodecanedioic acid or a derivative thereof from a biological source is disclosed. Another aspect of the invention relates to a composition comprising a reproducible isolated 3-hexenedioic acid and its 3-hexenedioic acid derivative. In some embodiments, the biological source-derived 3-hexenedioic acid and its 3-hexenedioic acid derivative are characterized by a ¹⁴ C / ¹² C isotope ratio of about 1.2 × 10 ⁻¹² . A further aspect of the invention relates to a composition comprising a polyamide derived from a biological source. In some embodiments, the polyamide is a nylon 6,12 polymer and is derived from a renewable carbon source with at least 12 carbon atoms per monomer unit. In some embodiments, the nylon 6,12 includes a detectable trace of carbon 14. In some embodiments, the renewable compounds disclosed herein comprise up to about 1 trillionth carbon 14.

  Aspects of the invention relate to compositions comprising dodecenedioic acid or dodecenedioic acid derivatives, and at least one unsaturated dicarboxylic acid or unsaturated dicarboxylic acid derivative byproduct obtained from the dodecenedioic acid or dodecenedioic acid derivative. . In some embodiments, the composition comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8 derived from the dodecenedioic acid or dodecenedioic acid derivative. Seeds, containing at least 9 by-products. In some embodiments, the at least one unsaturated dicarboxylic acid or unsaturated dicarboxylic acid derivative byproduct comprises an alkene chain having from about 7 to about 16 carbon atoms. In some embodiments, the composition comprises at least nine byproducts obtained from the dodecenedioic acid or dodecenedioic acid derivative, wherein the byproducts contain about 7 to about 16 carbon atoms. Containing alkene chains. In some embodiments, the dodecenedioic acid derivative is a dodecenedioic acid diester. In a preferred embodiment, the dodecenedioic acid or dodecenedioic acid derivative contains up to about 1 trillion carbon 14. Preferably, the alkene chain contains a carbon double bond at the C3-C4 position.

  Another aspect of the present invention is the use of 9-octadecenedioic acid or 9-octadecenedioic acid derivative and at least one octadecenedioic acid or octadecenedioic acid derivative secondary product obtained from 9-octadecenedioic acid or 9-octadecenedioic acid derivative. It relates to a composition comprising the product. In some embodiments, the composition is at least one, at least 3, at least 4, at least 5, at least 6, at least 7 derived from 9-octadecenedioic acid or 9-octadecenedioic acid derivatives. , At least 8, at least 9, at least 10, at least 15 and at least 16 by-products. In some embodiments, the octadecenedioic acid or octadecenedioic acid derivative by-product is C1-C2, C2-C3, C3-C4, C4-C5, C5-C6, C6-C7, C7-C8, C8. It contains a carbon double bond at the position C9, C10-C11, C11-C12, C12-C13, C13-C14, C14-C15, C15-C16, C16-C17, or C17-C18. In some embodiments, the 9-octadecenedioic acid or 9-octadecenedioic acid derivative comprises up to about 1/1 trillion carbon 14.

  The advantages of the invention described above, together with further advantages, may be understood by reference to the following description, taken in conjunction with the accompanying drawings. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of the invention.

FIG. 1 shows a general pathway for aromatic amino acid biosynthesis and a divergent pathway for synthesizing cis, cis-muconic acid from 3-dehydroshikimate. FIG. 1 shows a general pathway for aromatic amino acid biosynthesis and a branched pathway for synthesizing cis, cis-muconic acid from 3-dehydroshikimate. FIG. 2 shows a process flow schematic diagram of an embodiment of dodecanedioic acid synthesis. FIG. 2 shows a self-metathesis reaction to form a symmetric Δ ⁹ octadecenedioic acid followed by a cross-metathesis reaction with 3-hexenedioic acid. FIG. 3 shows the effect of double bond transfer during the self-metathesis reaction. FIG. 4 shows the effect of double bond migration during the cross metathesis reaction.

(Detailed description of the invention)
Aspects of the invention relate to methods and compositions for producing dodecenedioic acid and its derivatives and / or dodecanedioic acid and its derivatives. In some aspects, the present invention relates to methods and compositions for producing dodecenedioic acid from 3-hexenedioic acid and octadecenedioate. In a preferred embodiment, dodecenedioic acid is generated from dimethyl hexenedioate and dimethyl octadecenedioate. In a preferred embodiment, the reduction of dimethyl dodecenedioate produces dimethyl dodecanedioate. As used herein, the terms hexenedioic acid and hexanedioate are used interchangeably and include 6 carbon atoms, 8 hydrogen atoms, and 4 oxygen atoms, A molecule having the formula: HOOC—CH ₂ —CH═CH—CH ₂ —COOH. As used herein, the terms dodecenedioic acid and dodecenedioate are used interchangeably and include 12 carbon atoms, 20 hydrogen atoms, and 4 oxygen atoms, having the formula: HOOC A molecule having —CH ₂ —CH═CH— (CH ₂ ) ₇ —COOH. As used herein, the terms dodecanedioic acid and dodecanedioic acid are used interchangeably and include 12 carbon atoms, 22 hydrogen atoms, and 4 oxygen atoms, A molecule having the formula: HOOC— (CH ₂ ) ₁₀ —COOH.

  Several multi-step chemical processes have been used to produce dodecanedioic acid, usually from cyclohexanone, cyclododecadiene or cyclododecatriene. Dodecanedioic acid uses hydrogen peroxide and acetic acid to form its corresponding epoxy compound, which is later hydrogenated to form an alcohol and oxidized to form the desired product. 1,5,9 can be produced from the epoxidation of cyclodecatriene. In another chemical process, the dodecanedioic acid is epoxidized with 1,5,9 cyclodecatriene with organic hydroperoxide to form 1,2-epoxy-5,9-cyclododecadiene, after which the compound Is converted to a cyclo derivative that is oxidizable to dodecanedioic acid. In some aspects, the present invention uses oils or fats and muconic acid as alternative starting materials for the production of dicarboxylic acids, oxo compounds (eg, oxoaldehydes and oxoesters).

  As used herein, the phrases “preferred” and “preferably” refer to embodiments of the invention that provide certain benefits under certain circumstances. However, other embodiments may also be preferred under the same or other circumstances. Furthermore, the description of one or more preferred embodiments does not imply that other embodiments are not useful and is not intended to exclude other embodiments from the scope of the invention.

  The singular forms “a”, “a”, “an”, and “above, the” refer to the plural unless the context clearly indicates otherwise. Includes mention.

  The terms “including, including” and “including, including” are used in the sense of a comprehensive open system, meaning that further elements can be included.

  The term “including” is used to mean “including but not limited to”. “Including, including, including” and “including but not limited to” are used interchangeably.

  As used herein, the following terms and phrases shall have the meanings set forth below. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

  Any numerical value described herein includes all values from the lower value to the upper value. All possible combinations of numerical values between the lower limit value and the upper limit value recited herein are expressly included in the present application.

  As used herein, the term “aldehyde” refers to a carbonyl-containing functional group having the formula:

Here, R is substantially any group, and examples include, but are not limited to, aliphatic, substituted aliphatic, aryl, arylalkyl, heteroaryl and the like.

As used herein, the term “aliphatic” refers to a substantially hydrocarbon-based compound, or a radical thereof (eg, C ₆ H ₁₃ for a hexane radical), where alkanes, alkenes, and alkynes are And includes straight chain and branched chain configurations, as well as all stereoisomers and positional isomers.

As used herein, the term “alkyl” refers to a hydrocarbon arranged in a chain in a homologous series having the general formula: C _n H _{2n + 1} . Examples of the alkyl substituent include methyl (CH ₃ —), ethyl (C ₂ H ₅ —), propyl (C ₃ H ₇ —), butyl (C ₄ H ₉ —), pentyl (C ₅ H ₁₁ —) and the like. Can be mentioned. The structure of an alkyl group is similar to that of its alkane counterpart, but has one fewer hydrogen atom.

As used herein, the term “aryl” refers to a substantially hydrocarbon-based fragrance as a substituent attached to another group having a ring structure (specifically, another organic group). Group compounds or radicals thereof (for example, C ₆ H ₅ ), and include benzene, naphthalene, phenanthrene, anthracene, and the like.

As used herein, the term “arylalkyl” refers to a substituent attached to another group (specifically, another organic group) that includes both aliphatic and aromatic structures. Or a radical thereof (for example, C ₇ H ₇ for toluene).

  As used herein, the term “carboxylic acid” is a compound having the formula: R—COOH, where R is substantially any group (for example, aliphatic, substituted, , Aliphatic, aryl, arylalkyl, heteroaryl, and the like, which are not limited thereto.

  As used herein, the term “cyclic” refers to a substantially hydrocarbon ring-closed compound or radical thereof. Cyclic compounds or substituents may also contain one or more sites of unsaturation. Exemplary cyclic compounds typically include compounds having 3 or more carbon atoms in the ring, more typically 4 or more, and even more typically 5 or more carbon atoms. And cyclopentane, cyclopentadiene, cyclohexene, cyclohexadiene, and conjugated derivatives thereof (eg, compounds having olefins conjugated with carbonyl functional groups (eg, carboxylic acids), amides and esters). However, it is not limited to these.

  As used herein, the term “derivative” refers to a molecule that differs in chemical structure from the parent compound. Examples of derivatives include homologues that are incrementally different in chemical structure from the parent (eg, aliphatic chain length differences); molecular fragments; structures that differ from the parent compound by one or more functional groups (eg, By changing one or more functional groups of the parent (eg, by changing the acid functionality of the parent molecule to an acid halide, amide, or ester); changing the ionization state of the parent (eg, Ionize the acid to its conjugate base); isomers (including positional isomers, geometric isomers, and stereoisomers); and combinations thereof, including but not limited to.

  As used herein, the term “ester” refers to a compound having the following formula:

Here, R and R ′ are independently selected from virtually any group including aliphatic, substituted aliphatic, aryl, arylalkyl, heteroaryl, and the like.

  As used herein, the term “heteroaryl” refers to an aromatic ring-closed compound or radical thereof as a substituent attached to another group (specifically, another organic group). In the above aromatic ring, at least one atom is other than carbon (for example, oxygen, sulfur and / or nitrogen).

  As used herein, the term “heterocyclic” refers to cyclic (ie, closed) as a substituent attached to another group (specifically, another organic group). An aliphatic compound or a radical thereof, wherein at least one atom in the ring structure is other than carbon (for example, oxygen, sulfur and / or nitrogen).

  As used herein, the term “ketone” refers to a compound having the formula:

Here, R and R ′ are independently selected from virtually any group (including but not limited to aliphatic, substituted aliphatic, aryl, arylalkyl, heteroaryl, etc.).

  As used herein, the term “lower” organic compound refers to an organic compound or radical thereof having 10 or fewer carbon atoms in the chain (including all branching or stereochemical variations). Specific examples include methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, and decyl.

  As used herein, the term “substituted” typically refers to a basic compound (eg, fatty acid) in which a second atom, substituent, functional group, and the like are attached in place of a hydrogen atom. Family, aryl, arylaliphatic, heterocyclic, heteroaryl, or heteroarylaliphatic compounds) or radicals thereof. For example, a substituted aryl compound or substituent can have an aliphatic group attached to the aryl-based closed ring (eg, for toluene, it has a substituted methyl group instead of a benzene hydrogen atom) . Again, simply and without limitation, long chain hydrocarbons include, without limitation, atoms or substituents attached thereto (eg, halide, heteroatom, functional group, aryl group, cyclic group, heteroaryl Group or heterocyclic group).

  As used herein, the term “unsaturated fatty acid” refers to a compound having an alkene chain and a terminal carboxylic acid group.

  As used herein, the term “unsaturated dicarboxylic acid” refers to each end of an unbranched carbon chain (which also contains at least one double bond in the carbon chain). A compound having a carboxylic acid.

(Muconic acid)
The description of the present invention uses the terms “muconate” and “muconic acid”. The term “muconic acid” refers to a chemical species in which both carboxylic acid functional groups are protonated and the molecule is formally a neutral species. The term “muconate” means that one or both of the carboxylic acid functional groups are deprotonated to form an anionic or doubly anionic form of muconic acid (a major species at physiological pH values). A chemical species that gives a certain). However, since the terms “muconic acid” and “muconate” refer to the protonated or deprotonated form of the same molecule, the term refers to the protonated and deprotonated forms of the molecule (eg, non-ionized and ionized forms). Used as a synonym when the difference between (form) is not usefully distinguished.

  Hexa-2,4-dienedioic acid has three isomers (generally referred to as muconic acid: these are trans, trans (2E, 4E) isomer, cis, trans (2Z, 4E ) And cis, cis (2Z, 4Z) isomers):

Cis cis-muconic acid and trans, trans-muconic acid are commercially available in small quantities (eg, from Sigma-Aldrich) but are very expensive. However, cis, cis-muconic acid is also produced by enzymatic degradation of aromatic compounds by several bacteria. In some embodiments, cis, cis-muconic acid is disclosed in US Pat. Nos. 5,487,987 and 5,616,496, which are hereby incorporated by reference. As is biologically synthesized by certain bacteria. Industrial scale quantities of cis, cis-muconic acid can be produced by such biosynthesis. Bacteria that can biosynthesize cis, cis-muconic acid are members of a genus that have an endogenous common pathway of aromatic amino acid biosynthesis. Examples of such bacteria include the genus Escherichia, Klebsiella, Corynebacterium, Brevibacterium, Arthrobacter, Bacillus, Pseudomonas, Streptomyces, Staphylococcus, or the genus Sraphylococcus. Eukaryotic host cells may also be utilized, particularly yeasts of the genus Saccharomyces or Schizosaccharomyces.

  Suitable prokaryotic species, Escherichia coli, Klebsiella pneumonia, Corynebacterium glutamicum, Corynebacterium herculis, Brevibacterium divaricatum, Brevibacterium lactofermentum, Brevibacterium flavum, Bacillus brevis, Bacillus cereus, Bacillus circulans, Bacillus coagulans, Bacillus lichenformis, Bacillus megaterium, Bacillus mesentericus, Bacillus pumilis, Bacil us subtilis, Pseudomonas aeruginosa, Pseudomonas angulata, Pseudomonas fluorescens, Pseudomonas tabaci, Streptomyces aureofaciens, Streptomyces avermitilis, Streptomyces coelicolor, Streptomyces griseus, Streptomyces kasugensis, Streptomyces lavendulae, Streptomyces lipmanii, Streptomyces Iividans, Staphylococcus epidermis, Staphylococcus saprophyti us, and Serratia marcescens, and the like. Suitable eukaryotic species include Saccharomyces cerevisiae and Saccharomyces carlsbergensis.

  In some embodiments, the method includes microbial biosynthesis of biogenic muconic acid from a commercially renewable carbon source (see, eg, US Pat. No. 5,616,496, which is herein incorporated by reference). Incorporated by reference). As used herein, the term “biologically derived” refers to a material obtained using a biological process as opposed to a non-biological process (eg, a chemical process by synthesis). Mention. For example, “biologically derived” muconic acid is obtained from a fermentation process that utilizes a fermentable carbon source. A “biologically derived” compound or product refers to a product that is composed in whole or in part of material derived from biological sources. In some embodiments, preferred host cells for use in the present invention can convert the carbon source to D-erythrose-4-phosphate (E4P) and phosphoenolpyruvate (PEP). In some embodiments, E4P and PEP are then converted to amino acids via metabolic pathways that ultimately produce aromatic amino acids. Fermentable carbon sources can be biocatalyzed to D-erythrose 4-phosphate (E4P) and phosphoenolpyruvate (PEP), two precursors to a common pathway of aromatic amino acid biosynthesis Essentially any carbon source can be included. Suitable carbon sources include biomass-derived renewable resources (eg starch, cellulose, polyols (eg glycerol), pentose sugars (eg arabinose and xylose) hexose sugars (eg glucose), and fructose, disaccharides ( For example, but not limited to, sucrose and lactose), and other carbon sources that can support microbial metabolism, such as, but not limited to, glucose, glycerol, sucrose, xylose and In one embodiment, D-glucose is a biomass-derived carbon source.

  Suitable host cells for use in the present invention include members of a genus that can be utilized for the biological production of desired intermediates in the biosynthesis of aromatic compounds (eg, amino acids). In some embodiments, such host cells are suitable for industrial scale biosynthesis or biological production of industrially useful aromatic compounds and intermediates that provide such useful compounds. . One intermediate in the pathway of aromatic amino acid biosynthesis is 3-dehydroshikimate (DHS). In particular, a suitable host cell may have an endogenous common pathway of aromatic amino acid biosynthesis that functions at least for the production of DHS. A common pathway for the biosynthesis of aromatic amino acids is endogenous in a wide variety of microorganisms and can be used to produce a variety of aromatic compounds. In certain embodiments, auxotrophic cell lines with mutations that block the conversion of 3-dehydroshikimate (DHS) to chorismate are used. Such mutants have mutations in one or more of the genes encoding shikimate dehydrogenase, shikimate kinase, EPSP synthase or chorismate synthase. These mutants accumulate elevated intracellular levels of DHS. Suitable mutant cell lines include Escherichia coli AB2834 strain, AB2829 strain, and AB2849 strain.

  For example, E.I. E. coli AB2834 has a mutation at the aroE locus encoding shikimate dehydrogenase, which prevents the cells from converting DHS to shikimate. As a result, the carbon flow directed towards aromatic amino acid biosynthesis does not advance beyond DHS. Similarly, E.I. E. coli AB2829 accumulates DHS. Because it is not possible to convert shikimate 3-phosphate (S3P) to 5-enolpyruvylshikimate-3-phosphate (EPSP) due to a mutation in the aroA locus encoding EPSP synthase. . E. E. coli AB2849 cannot catalyze the conversion of EPSP to chorismate due to mutations in the aroC locus encoding chorismate synthase. This also results in an increase in intracellular levels of DHS.

  Host cells can be transformed such that intracellular DHS can be used as a substrate for biocatalytic conversion to catechol, which can then be converted to muconic acid. For example, host cells can be transformed with recombinant DNA to divert carbon flow away from the common pathway of aromatic amino acid biosynthesis after DHS is produced and into a branch pathway to produce muconic acid. You can let it enter.

  Mechanisms for transforming host cells to direct the flow of carbon into the branching pathway include expression encoding 3-dehydroshikimate dehydratase, protocatechuate decarboxylase, and catechol 1,2-dioxygenase. It may include the insertion of genetic elements containing possible sequences. Regardless of the exact mechanism utilized, expression of these enzymatic activities is expected to be effected or mediated by the transfer of recombinant genetic elements into the host cell. As defined herein, a genetic element is a product (eg, protein, apoprotein, or antisense RNA that can be implemented or controlled to enzymatically function the pathway) A nucleic acid (generally DNA or RNA) having an expressible coding sequence for the protein, wherein the expressed protein can function as an enzyme, can inhibit enzyme activity, or can be activated by releasing the inhibition. The nucleic acid encoding these expressible sequences can be chromosomal (eg, integrated into the host cell chromosome) or extrachromosomal (eg, maintained by a plasmid, cosmid, etc.). Can be either.

  The genetic elements of the invention can be introduced into a host cell by a plasmid, cosmid, phage, yeast artificial chromosome, or other vector that mediates transfer of the genetic element into the host cell. These vectors may contain an origin of replication along with the vector and cis-acting control elements that control the replication of the genetic element carried by the vector. A selectable marker can be present in the vector to assist in identifying the host cell into which the genetic element has been introduced. For example, the selectable marker can be a gene that confers resistance to a particular antibiotic (eg, tetracycline, ampicillin, chloramphenicol, kanamycin, or neomycin).

  Introducing a genetic element into a host cell can utilize an extrachromosomal multicopy plasmid vector into which the genetic element is inserted. Induction of the genetic element into the host cell provided by the plasmid involves the initial cleavage of the plasmid with a restriction enzyme followed by ligation of the plasmid with the genetic element according to the invention. Upon recirculation of the ligated recombinant plasmid, transduction of other mechanisms for plasmid transfer (eg, electroporation, microinjection, etc.) is required to transfer the plasmid into the host cell. Used. Suitable plasmids for inserting genetic elements into the host cell include pBR322 and its derivatives (eg, pAT153, pXf3, pBR325, pBr327, pUC vector), pACYC and its derivatives, pSC101 and its derivatives, and ColE1 However, it is not limited to these. In addition, cosmid vectors (eg, pLAFR3) are also suitable for insertion of genetic elements into host cells. Examples of plasmid constructs include p2-47, pKDS. 243A, pKD8.243B, and pSUaroZY157-27, which have the aroZ and aroY loci isolated from Klebsiella pneumoniae (encoding 3-dehydroshikimate dehydratase and protocatechuate decarboxylase, respectively) ), But is not limited thereto. Additional examples of plasmid constructs include pKDS. 292, which has a gene fragment endogenous to Acinetobacter calcoaceticus catA encoding catechol 1,2-dioxygenase.

  Methods for transforming host cells can also include insertion of genes encoding enzymes that increase carbon donation to a common pathway for aromatic amino acid biosynthesis. Gene expression is primarily directed by its own promoter, but other genetic elements (including optional expression control sequences (eg, repressors and enhancers)) are proteins, apoproteins, or anti-antigens. It can be included to control the expression or repression of the coding sequence of the sense RNA. In addition, recombinant DNA constructs can be generated by replacing the natural promoter of the gene with an alternative promoter to increase expression of the gene product. Promoters can be either constitutive or inducible. Constitutive promoters control gene transcription at a constant rate during the life of a cell, whereas inducible promoter activity varies as determined by the presence (or absence) of a particular inducer . For example, control sequences can be inserted into wild-type host cells to facilitate overexpression of selected enzymes already encoded in the host cell genome, or alternatively, synthesis of enzymes encoded extrachromosomally. Can be used to control

  Regulatory sequences can be used to promote overexpression of DHS. As previously indicated, DHS is encoded by 3-deoxy-D-arabinohepturonic acid 7-phosphate (DAHP) synthase (encoded by aroF) along with transketolase (encoded by tkt), which is a pentose phosphate pathway enzyme. ) And 3-dehydroquinate (DHQ) synthase (encoded by aroB) are synthesized in a common pathway by a series of catalytic activities of tyrosine-sensitive isozymes. The expression of these biosynthetic enzymes can be amplified to increase the conversion of D-glucose to DHS. Increasing the in vivo catalytic activity of DAHP synthase (which is the first enzyme of the common pathway) increases the flow of D-glucose equivalents directed towards aromatic biosynthesis. However, although some level of DAHP synthase catalytic activity is achieved, beyond this, no further improvement is achieved in the proportion of D-glucose conferred on aromatic biosynthesis. At this limiting level of aromatic amino acid biosynthesis, catalytic level amplification of the pentose phosphate pathway enzyme, transketolase, achieves a significant increase in the proportion of D-glucose drawn up into the pathway.

  Amplified transketolase activity can increase D-erythrose 4-phosphate concentration. Limited D-erythrose 4-phosphate availability as one of the two substrates of DAHP synthase may limit DAHP synthase catalytic activity. Accordingly, one method for increasing the catalytic activity of DAHP synthase, DHQ synthase and DHQ dehydratase is to overexpress the enzyme species by transforming the microbial catalyst with recombinant DNA sequences encoding these enzymes. It is.

  Amplified expression of DAHP synthase and transketolase can create a carbon flow surge directed to a common pathway of aromatic amino acid biosynthesis (more than the normal carbon flow directed to this pathway) . If the conversion rate of individual substrates to products catalyzed by individual enzymes in the common aromatic amino acid pathway is lower than the rate of DAHP synthesis, these rate-limiting enzyme substrates can accumulate in the cell.

  Microorganisms (eg, E. coli) successfully deal with accumulated substrates by exporting such substrates to their external environment (eg, bulk fermentation medium). This results in a loss of carbon flow from the common path. This is because the exported substrate typically does not undergo metabolism of the microorganism. DHQ synthase is an example of a rate limiting common pathway enzyme. Amplified expression of DHQ synthase removes the rate-limiting feature of the enzyme and prevents the accumulation of DAHP, its non-phosphorylated analog, DAH. DHQ dehydratase is not rate limiting. Thus, amplified expression of aroF-encoded DAHP synthase, tkt-encoded transketolase and aroB-encoded DHQ synthase increases the production of DHS, which is a DHS dehydratase and protocatechu It is converted to catechol in the presence of ate decarboxylase, which is then converted to cis, cis-muconic acid by biocatalysis.

  Accordingly, as a preferred embodiment of the present invention, an atypical strain of Escherichia coli expressing genes encoding DHS dehydratase, protocatechuate decarboxylase, and catechol 1,2-dioxygenase was constructed. Biocatalytic conversion of D-glucose to acid has become possible. Efficient conversion of D-glucose to DHS was achieved upon transformation of the host cells with pKD136. The strain E. E. coli AB2834 / pKD136 was transformed into plasmids pKD8.243A and pKDS. 292 was transformed. The result was that E. coli expressing the above enzymes, 3-dehydroshikimate dehydratase (aroZ), protocatechuate decarboxylase (aroY) and catechol 1,2-dioxygenase (catA). coli AB2834 / pKD136 / pKDS. 243A / pKDS. 292. This bacterial cell line was deposited with the American Type Culture Collection (12301 Parklawn Drive, Rockville MD 20852) on August 1, 1995 and assigned accession number 69875.

  In another embodiment, E.I. E. coli AB2834 / pKD136 was transformed with plasmids p2-47 and pKD8.292. coli AB2834 / pKD136 / p2-47 / pKDS. 292 is generated. In another embodiment, E.I. E. coli AB2834 / pKD136 is the plasmid pKD8.243B and pKDS. Transformed with E. 292; coli AB2834 / pKD136 / p2-47 / pKDS. 292 is generated. Each of these atypical host cell lines catalyzes the conversion of D-glucose to cis, cis-muconic acid. The synthesized cis, cis-muconic acid accumulates extracellularly and can be separated from the cells. The cis, cis-muconic acid can then be isomerized to cis, trans-muconic acid and, if desired, further to trans, trans-muconic acid.

  Accordingly, some aspects of the invention relate to transformants of host cells that have an endogenous common pathway of aromatic amino acid biosynthesis. The transformants are characterized by constitutive expression of heterologous genes encoding 3-dehydroshikimate dehydratase, protocatechuate decarboxylase, and catechol 1,2-dioxygenase. In one embodiment, the cell transformant is further transformed with an expressible recombinant DNA sequence encoding the enzyme, transketolase, DAHP synthase, and DHQ synthase. In another embodiment, the host cell is selected from the group of mutant cell lines, including mutations having mutations in a common pathway of amino acid biosynthesis that block the conversion of 3-dehydroshikimate to chorismate. In yet another embodiment, the genes encoding 3-dehydroshikimate dehydratase and protocatechuate decarboxylase are endogenous to Klebsiella pneumoniae. In a further embodiment, the heterologous gene encoding catechol 1,2-dioxygenase is endogenous to Acinetobacter calcaceticus.

  As shown in FIG. 1, the intermediates in the branched pathway are protocatechuate, catechol, and cis, cis-muconic acid. The enzyme responsible for biocatalytic conversion of DHS into protocatechuate is the enzyme 3-dehydroshikimate dehydratase (labeled “aroZ” in FIG. 1). The enzyme responsible for the decarboxylation of protocatechuate to form catechol is protocatechuate decarboxylase (indicated as “aroY” in FIG. 1). Finally, the enzyme that catalyzes the oxidation of catechol to produce cis, cis-muconic acid is catechol 1,2-dioxygenase (labeled “catA” in FIG. 1). According to standard notation, the genes for expression of these enzymes are shown using italics, so are aroZ, aroY, and catA, respectively. The cis, cis-muconic acid can then be isomerized (not shown). In one embodiment of the invention, the host cell may exhibit constitutive expression of the genes, aroZ, aroY, and catA. In another embodiment, the host cell may exhibit constitutive expression of any one or more of the above genes, aroZ, aroY and catA; or any combination of the two. In yet another embodiment, the host cell may not show any constitutive expression of aroZ, aroY and catA.

  The above enzymes, 3-dehydroshikimate dehydratase and protocatechuate decarboxylase are microorganisms (e.g., Neurospora, Aspergillus, Acinetobacter, Klebsiella, and Pseudomonas) are aromatic (benzoate and p-hydroxybenzoate) and hydroaromatic (shikimate). And quinate) are supplemented from an ortho-cleavage pathway that can be used as the sole carbon source for growth. DHS dehydratase plays an important role in the microbial catabolism of quinic acid and shikimic acid. Protocatechuate decarboxylase was formulated by Patel to catalyze the conversion of protocatechuate to catechol during catabolism of p-hydroxybenzoate by Klebsiella aerogenes. Retest of Patel's strain (currently referred to as Enterobacter aerogenes) [(a) Grant, D. et al. J. et al. W. Patel, J .; C. Antoni van van Leewenhoek 1969, 35, 325. (B) Grant, D.D. J. et al. W. Antoni van van Leewenhoek 1970, 36, 161], recently, Ornston concluded that protocatechuate decarboxylase was not metabolically important in the catabolism of p-hydroxybenzoates [Doten, R .; C. Ornston, N .; J. et al. Bacteriol. 1987, 169, 5827].

(Renewable compounds)
Compounds of interest containing at least one carbon atom (eg, muconic acid, dodecanedioic acid, dodecenedioic acid, 3-hexenedioic acid and their derivatives generated from renewable biologically derived carbon sources) Is composed of carbon derived from atmospheric carbon dioxide incorporated by plants (eg, derived from a carbon source (glucose, sucrose, glycerin, or vegetable oil)). Accordingly, such compounds contain renewable carbon in their molecular structure rather than fossil fuel-based or petroleum-based carbon. Thus, dodecanedioic acid or products derived from dodecanedioic acid derived from the above biological sources, and related derivative products have a smaller carbon footprint than dodecanedioic acid and related products generated by conventional methods. Because they do not deplete fossil fuels or oil reserves, and do not increase the amount of carbon in the carbon cycle (eg, life cycle analysis does not show a net increase in carbon balance to the global carbon balance).

The dodecanedioic acid and related products from the above biological sources, as well as their starting compounds (eg, muconic acid) or intermediate products (eg, 3-hexenedioic acid) can be prepared by methods (eg, dual) known in the art. It can be distinguished from products produced from fossil fuels or petrochemical carbon sources by dual carbon-isotopic finger printing. This method can distinguish materials that have been chemically identified by another method, using ¹⁴ C and ¹³ C isotope ratios in the material by biological versus non-biological sources. Distinguish between carbon atoms. The carbon isotope ¹⁴ C is unstable and has a half-life of 5730 years. The relative amount of the unstable ¹⁴ C isotope relative to the stable ¹³ C isotope is measured between the fossil (dead and long) feed and the biosphere (live and therefore renewable) feed. The sample carbon can be distinguished (Currie, LA, “Source Application of Atmospheric Particles,” Characteristic of Environmental Particles, J. Buffel and H. ven L. e. IUPAC Environmental Analytical Chemistry Series (Lewis Publishers, Inc. (1992) 3-74). The basic assumption in radiocarbon dating is that atmospheric ¹⁴ C concentration continuity results in ¹⁴ C continuity in living organisms.

When dealing with an isolated sample, the age of the sample can be approximately derived by the following relationship: t = (− 5730 / 0.693) ln (A / A _o ) where t = age. 5730 is the half-life of the unstable ¹⁴ C isotope, and A and A _o are the ¹⁴ C radioactivity of the sample and modern standards, respectively (Hsieh, Y., Soil ScL Soc. Am J., 56, 460, (1992)). However, due to atmospheric nuclear testing since 1950 and the burning of fossil fuel since 1850, ¹⁴ C gained a second geochemical time feature. Its concentration in atmospheric CO ₂ , and therefore in the living biosphere, has been about twice that of the peak of the nuclear test (mid 1960s). The concentration in the atmospheric CO ₂ has since been estimated to be about 1.2 × 10 ⁻¹² steady state cosmic ray (atmosphere) with an approximate relaxation half-life of 7-10 years since then. ) Gradually returning to the baseline isotope ratio ( ¹⁴ C / ¹² C) (this former half-life must be distinguished from the isotope half-life (ie, in the atmosphere and since the beginning of the nuclear age) A detailed atmospheric nuclear input / decay function must be used to track ¹⁴ C variations in the biosphere)). Its concentration in atmospheric CO ₂ is a ¹⁴ C time characteristic in this latter biosphere that provides the prospect of recent biosphere carbon dating. ¹⁴ C can be measured by accelerator mass spectrometry (AMS), and the results are expressed in units of fractions of modern carbon (f _M ). f _M is defined by National Institutes of Standards and Technology (NIST) Standard Reference Materials (SRMs) 4990B and 4990C (known as oxalic acid standards HOxI and HOxII, respectively). The above basic definition relates to 0.95 × ¹⁴ C / ¹² C isotope ratio HOxI (referred to AD 1950). For the current living biosphere (plant material), f _M ≈1.1.

The ratio of stable carbon isotopes ¹³ C and ¹² C provides a complementary means for resource differentiation and allocation. The ¹³ C / ¹² C ratio in a given biological source-derived material is the result of the ¹³ C / ¹² C ratio in atmospheric carbon dioxide at the time when carbon dioxide is fixed and also reflects the exact metabolic pathway. Regional variations also exist. Oil, in _{C 3} plants (the broadleaf), _{C 4} plants (herbaceous), and marine carbohydrates (marine carbonate) are ^all marked difference and the ¹³ C value at 13 ^{C / 12} C (i.e., [delta] ¹³ C value) The corresponding difference is shown. The ¹³ C measurement scale was initially defined by zeroing with pee dee belemnite (PDB) limestone. Here, the values are given in parts per thousand deviation from this material. The δ ¹³ C value is in parts per thousand (per mil (abbreviated as ‰)) and is calculated as follows:
δ ¹³ C = ( ¹³ C / ¹² C) sample− ( ¹³ C / ¹² C) standard substance / ( ¹³ C / ¹² C) standard substance × 1000 ‰.

Since the PDB reference material (RM) has been used up, a series of alternative RMs have been developed in cooperation with IAEA, USGS, NIST and other selected international isotope laboratories. The notation of permill deviation from PDB is δ ¹³ C. Measurements, with respect to CO _2, is carried out by a high precision stable density mass analysis of molecular ions of masses 44, 45 and 46 (IRMS). Furthermore, lipid matter of C ₃ plants and C ₄ plants, different analyzing the results derived from the carbohydrate components of the same plant material of the metabolic pathway. Within the accuracy of the measurement, ¹³ C shows a large variation due to the isotope fractionation effect. The most prominent of these is the photosynthesis mechanism in the context of the present invention. The main cause of the difference in the carbon isotope ratio in the plant is the difference in the pathway of photosynthetic carbon metabolism in the plant, in particular the reaction that occurs during the initial carboxylation (eg the initial fixation of atmospheric CO ₂ ). Closely related. Two major classes of vegetation, as incorporating a _{C 3} (or Calvin-Benson) photosynthetic pathway, those incorporating a _{C 4} (or Hatch-Slack) photosynthetic pathway. C ₃ plants (e.g., hardwood and softwood) is dominant in the temperate climate zones. The C ₃ plants, the primary CO ₂ fixation or carboxylation reaction, an enzyme, ribulose-1,5-diphosphate carboxylase required tosylate, first stable product is a compound of 3 carbons. In contrast, C ₄ plants include tropical herbaceous plants such as corn and sugar cane. In C ₄ plants, further carboxylation reaction requiring phosphoenolpyruvate carboxylase which is another enzyme, a major carboxylation reaction. The first stable carbon compound is a four carbon acid which is then decarboxylated. CO ₂ released in this way are re-fixed by the C ₃ pathway. Both C ₄ and C ₃ plants show a range of ¹³ C / ¹² C isotope ratios, but typical values are about -10 to -14 permil (C ₄ ) and -21 to -26 permil ( C ₃ ) (Weber et al., J. Agric. Food Chem., 45, 2942 (1997)). Coal and oil generally fall within this latter range.

Thus, compositions comprising muconic acid, dodecanedioic acid, and dodecanedioic acid derived from biological sources of the present invention are based on ¹⁴ C (f _M ) and dual carbon isotope fingerprinting and their ancient fossil fuels and It can be distinguished from its petrochemical counterpart. This represents a new composition (eg, US Pat. Nos. 7,169,588, 7,531,593, and 6,428,767). The ability to distinguish these products is beneficial in tracking these materials commercially. For example, products containing both new and old carbon isotope profiles can be distinguished from products made only from ancient materials. Thus, the biologically derived dodecanedioic acid and derivative materials can be tracked commercially based on their unique profiles.

Thus, a molecule or compound containing at least one carbon atom (e.g. muconic acid or derivative thereof, fatty acid or derivative thereof, 3-hexenedioic acid or derivative thereof, dodecenedioic acid or derivative thereof, dodecanedioic acid or derivative thereof, Dicarboxylic acids or derivatives thereof, polyamides or derivatives thereof, nylons or derivatives thereof) are described by their carbon isotope distribution (eg, ¹⁴ C, ¹³ C, or ¹² C) or by their ¹⁴ C / ¹² C ratio It is recognized that it can be done. Since each single carbon atom in a compound is derived from a naturally occurring carbon isotope, the source of carbon atoms affects the carbon isotope distribution of the compound or the ¹⁴ C / ¹² C ratio. . More specifically, the carbon isotope distribution or ^{14 C /} 12 ^C ratio of the compound synthesized from petrochemical feedstocks, the carbon isotope distribution or ¹⁴ of the compound produced from renewable carbon sources ^C / It can be distinguished from the ¹² C ratio. The basic assumption is that continuity of ¹⁴ C concentration in the atmosphere results in continuity of ¹⁴ C in living organisms, whereas in inorganic carbon sources (eg petrochemical feedstock) it is that all of the above ^{14 C} is accidentally collapse. Thus, compounds produced by renewable carbon sources can be distinguished from products produced from inorganic carbon sources. In some embodiments, the ¹⁴ C / ¹² C ratio is determined by ASTM test method D 6866-05, which uses radiocarbon and isotope ratio mass spectrometry to determine the bio-based contents of natural range materials. Which is incorporated herein by reference). Evaluation of the reproducibly forming carbon in the compound can be done via standard test methods. Radiocarbon and isotope ratio mass spectrometry can be used to determine the bio-based content of the material. ASTM International (formally known as American Society for Testing and Materials) has an established standard method for evaluating the bio-based contents of materials. The ASTM method is referred to as ASTM-D6866. This test method measures the ¹⁴ C / ¹² C isotope ratio in a sample, and ¹⁴ C / in the contents of a standard 100% biological source-derived material versus a given% biological source from the sample. Compare with ¹² C isotope ratio.

In some embodiments, the composition comprising biologically derived dodecanedioic acid, dodecenedioic acid, 3-hexenedioic acid, polyamide is renewable using a ¹⁴ C distribution or a ¹⁴ C / ¹² C ratio. Can be distinguished from the corresponding compounds which are not. In some embodiments, the ¹⁴ C / ¹² C ratio indicates a fraction of carbon atoms derived from a renewable carbon source. In an exemplary embodiment, the starting material to yield a dodecanedioic acid, muconic acid and delta ⁹ unsaturated fatty acids (e.g., oleic acid) is. In a preferred embodiment, muconic acid is generated from the carbon source biologically derived, the delta ⁹ unsaturated fatty acids are derived from plant sources or animal sources. Thus, dodecanedioic acid products obtained from cross-metathesis are derived from ancient carbons (eg, carbons derived from coal, oil or natural gas), all carbon atoms derived from renewable carbon sources. It has a ¹⁴ C / ¹² C ratio indicating no atoms. Synthesis of compounds from starting materials derived from renewable carbon sources that contain ¹⁴ C and ancient carbon sources that do not contain radioactive carbon is a ¹⁴ C / ¹² C ratio of compounds that are composed entirely of materials from biological sources. Result in a decrease in the ¹⁴ C / ¹² C ratio. The 1.2 × 10 ⁻¹² value represents the ¹⁴ C / ¹² C ratio of a compound composed entirely of materials derived from biological sources, and the 0 value is the ¹⁴ C / ¹² C ratio of a compound obtained from ancient carbon. By speculating that it corresponds, compounds that are partially formed from materials derived from biological sources have a radiocarbon signature or fingerprint lower than 1.2 × 10 ⁻¹² . In an exemplary embodiment, nylon 6,12 results from the condensation of 1,6-hexamethylenediamine and dodecanedioic acid. When 1,6-hexamethylenediamine is produced from a petrochemical feedstock (eg, from coal, petroleum, and natural gas) and dodecanedioic acid is all formed from a bio-derived material, 2/3 of the carbon atom content is derived from a renewable carbon source. In certain embodiments, the ¹⁴ C / ¹² C ratio of the desired product is greater than 0 or greater than 0.9 × 10 ⁻¹² . In some embodiments, the biogenic carbon content of the compound can be analyzed, and the ratio of the amount of ¹⁴ C is reported relative to that of the biosource-derived reference standard (percent of modern carbon). obtain. In some embodiments, the carbon content of the product from the biological source is 100%, at least 90%, at least 80%, at least 70%, at least 60%, at least 50%, at least 40%, at least 30% is there.

  Aspects of the invention relate to products prepared by the processes described herein. In some embodiments where the starting muconic acid is prepared from biomass, the resulting product of the process comprises a significant proportion of carbon obtained from renewable resources. Such a product is unique. This is because the product contains a detectable trace or some amount of carbon 14 (preferably up to about 1 trillionth of a trillion) as determined according to ASTM D6866-08. The resulting product is preferably 3 or more carbons, more preferably 9 or more carbons, more preferably 12 derived from renewable resources (eg, biomass (preferably by microbial synthesis)). Contains more than one carbon. The resulting product is prepared from renewable resources prepared by microbial synthesis under fermenter controlled conditions. In embodiments where the product is utilized to prepare a polymer, the monomer unit preferably comprises 12 or more carbons (derived from renewable resources (eg, biomass)).

(Muconic acid derivative)
In some aspects of the invention, muconic acid is reduced to 3-hexenedioic acid. In some embodiments, cis, cis-muconic acid and derivatives thereof that are biologically produced by recombinant host cell culture are first reduced to 3-hexenedioic acid. In some embodiments, 3-hexenedioic acid is used in a metathesis reaction to form the desired compound. General formula 1 illustrating muconic acid and muconic acid derivatives within the scope of the present invention is provided below:

Referring to Formula 1, R ₁ -R ₆ are typically aliphatic, substituted aliphatic, alkoxy, amino, amine, substituted amine, protected amine, aryl, substituted aryl, Arylalkyl, substituted arylalkyl, carbonyl-containing moieties (eg, aldehydes, amides, carboxylic acids, esters, ketones and thioesters), cyclic, substituted cyclic, ether, substituted ether, halide, heteroaryl, Independent of substituted heteroaryl, heterocyclic, substituted heterocyclic, hydrogen, hydroxyl, hydroxylamine, and nitrogen-containing moieties (eg, nitrile (RCN), nitro (NO ₂ ) and nitroso (RNO)) Selected. Preferably, R _{1 to} R ₆ are independently selected from aliphatic, typically lower aliphatic, and even more preferably lower alkyl, and hydrogen. R ₁ and R ₆ are most typically independently hydrogen or lower alkyl. R _{2 to} R ₅ are most typically hydrogen. Formula 1, and certain other structural formulas provided herein, in contrast to the dashed lines, indicate that all possible stereoisomers are included in that particular general formula, Includes the coupling indicated by the wavy lines.

  In some embodiments, the main derivative made from muconic acid is (1) a conjugate base, (2) various stereoisomers produced by the isomerization reaction of the parent compound, and / or (3) 1 A compound formed by converting one or more carboxylic acid functional groups to another functional group. In some embodiments, the cis, cis-muconic acid is converted to trans, trans-muconic acid by dissolving the cis, cis-muconic acid in methanol with trace elements and exposing the reaction mixture to light. Can be isomerized. The methane solubility of cis, cis-muconic acid and trans, trans-muconic acid is substantially different. As a result, trans, trans-muconic acid precipitates out of solution as it forms.

  In some embodiments, the cis, cis-muconic acid can be isomerized to cis, trans-muconic acid using methods known in the art. Since the cis, trans-muconic acid is more soluble in both organic solvents and aqueous media than either the cis, cis-isomer or trans, trans-isomer, cis, trans-muconic acid is easily It should be recognized that this is an advantageous starting compound that allows for convenient processing and recovery.

  In various embodiments, the method comprises subjecting a recombinant cell expressing 3-dehydroshikimate dehydratase, protocatechuate decarboxylase and catechol 1,2-dioxygenase to a medium containing a renewable carbon source. The step of culturing under conditions where such a renewable carbon source is converted to DHS by an enzyme found in the common pathway of aromatic amino acid biosynthesis of the cells, the resulting DHS being cis, cis- Converted to muconic acid by biocatalyst. In certain embodiments, the fermentation broth is provided in a vessel, such as a fermentation vessel, and an isomerization reaction can be performed in the vessel.

  Production of cis, cis-muconic acid by fermentation of the renewable carbon source can produce a broth containing the recombinant cells and extracellular cis, cis-muconic acid. The production also separates the recombinant cells, cell debris, insoluble proteins and other undesired solids from the broth and causes substantially all or large amounts of cis, cis-muconic acid formed by the fermentation. A step of providing a clear fermentation broth containing portions may be included. After muconic acid is produced, it can accumulate in the extracellular medium (eg, fermentation broth) and can be separated from the cells by centrifugation, filtration, or other methods known in the art. In some embodiments, ic, cis-muconic acid is first isomerized to cis, trans-muconic acid, which is then precipitated, extracted, filtered from the fermentation broth or cell-free fermentation broth. Or by other methods known in the art.

In some aspects of the invention, the carboxylic acid functionality of the muconic acid is converted to a variety of different functional groups that facilitate the cycloaddition reaction. This transformation facilitates the desired physical properties of the compound made by the above cycloaddition reaction, or a derivative or polymer made therefrom using the compound produced by cycloaddition as the monomer, or both. Exemplary carboxylic acid functional group transformations include: acid halides (eg, thionyl chloride, phosphorous pentachloride or phosphorous pentabromide) known to those skilled in the art (eg, , Acid chloride) formation. The carboxylic acid functionality of the muconic acid can also be converted to an ester (eg, methyl ester or ethyl ester). Examples of methods for forming esters (especially lower alkyl esters) include the following: The alcohol / H ⁺ protocol (eg, methanol or ethanol and its corresponding methyl using methanolic sulfuric acid and the corresponding sulfuric acid). Form esters and ethyl esters (which are easy to perform but may be accompanied by double bond isomerization); methylate with diazomethane to form methyl esters; or iodize the acid in a social protocol Treatment with alkyl (eg MeI / Et ₄ NOH) In a preferred embodiment, the dimethyl ester derivative is obtained using dimethyl sulfate and potassium carbonate starting from cis, cis-muconic acid.

  Table 1 below provides a partial listing of exemplary muconic acid and lower alkyl muconic acid esters (particularly methyl muconic acid ester and ethyl muconic acid ester) and solvents useful for their melting point and purification by recrystallization.

(Reduction of muconic acid)
Scheme 1 below illustrates one embodiment of a method for reducing muconic acid or a muconic acid derivative to 3-hexenedioic acid or a derivative thereof (eg, a monoester or diester). As shown in Scheme 1, the muconic acid diene can be reduced to 3-hexenedioic acid using a zinc halide reagent in a suitable solvent (eg, pyridine). Kotora et al., Chem. Lett. , P. 236-237 (2000).

(Olefin metathesis)
As used herein, the term “olefin” refers to an unsaturated compound containing at least one carbon-carbon double bond. In an exemplary embodiment, the simplest olefin with only one double bond and no other functional group forms a homologous series of hydrocarbons having the general formula C _n H _2n . The term “lower olefin” refers to an organic compound having less than about 10 carbon atoms and containing at least one carbon-carbon double bond. Lower olefins can have one, two or more unsaturated bonds. Preferably, the lower olefin has one unsaturated bond. The lower olefin may be substituted with one or more substituents with the carbon chain at any position, provided that the one or more substituents are substantially inert with respect to the metathesis reaction. Suitable substituents include, but are not limited to, alkyl (preferably methyl), and hydroxy, ether, keto, and aldehyde.

  Olefin metathesis is an important reaction used in organic synthesis. Olefin metathesis is also known as a transalkylidene organic reaction, which entails cleavage of the alkene double bond, followed by redistribution of the alkylene fragment. The above reaction was carried out by Yves Chauvin, Richard R. Schrock and Robert H. et al. Developed by Grubbs, who shared the Nobel Prize in Chemistry in 2005. The olefin metathesis reaction proceeds in the presence of a catalytically effective amount of a metathesis catalyst. Exemplary metathesis catalysts include methylcarbene catalysts based on catalytic transition metals such as ruthenium, nickel, tungsten, osmium, chromium, rhenium, and molybdenum. Metathesis catalysts (which can be used in the process according to the invention) are all metathesis catalysts known to the person skilled in the art and suitable for metathesis reactions, or a mixture of at least two catalysts. In some embodiments, the olefin metathesis reaction uses a first generation Grubbs catalyst, a variant or derivative of the first generation Grubbs type catalyst. In other embodiments, the olefin metathesis reaction uses a second generation Grubbs catalyst, a variant or derivative of the Grubbs type catalyst. Examples of Grab's catalysts include benzylidene-bis (tricyclohexylphosphine) dichlororuthenium and benzylidene [1,3-bis (2,4,6-trimethylphenyl) -2-imidazolidinylidene] dichloro (tricyclohexylphosphine) ruthenium. For example, but not limited to. In other embodiments, a Schrock catalyst or a variant or derivative of the Schrock catalyst is used as a catalyst for the metathesis reaction. In still other embodiments, the catalyst is a Hoveyda-Grubbs catalyst or a modification of the Hoveyda-Grubbs catalyst. In some embodiments, at least one metathesis catalyst selected from carbene or carbyne complexes or mixtures of these complexes is used. The term “complex”, as used herein, refers to a metal atom and at least one ligand or complexing agent bound thereto. Metathesis catalysts are used using techniques known to those skilled in the art.

As shown in Scheme 2, 3-hexene diacid or derivative thereof (e.g., lower alkyl esters thereof) is an unsaturated dicarboxylic acid (in particular, delta ⁹ dicarboxylic fatty acids), are reacted in the presence of a metathesis catalyst To produce dodecenedioic acid. However, any suitable unsaturated fatty acid can be suitably used in the process of the present invention.

The alkene chain of the unsaturated fatty acid may be linear or branched, and may contain one or more functional groups in addition to the carboxylic acid group as necessary. For example, some carboxylic acids contain one or more hydroxyl groups. The alkene chain typically contains about 4 to about 30 carbon atoms, more typically about 4 to about 22 carbon atoms. In many embodiments, the alkene chain contains 18 carbon atoms (ie, C18 fatty acid). The unsaturated fatty acid has at least one carbon-carbon double bond in the alkene chain (ie, a monounsaturated fatty acid) and has more than one double bond in the alkene chain. To obtain (ie polyunsaturated fatty acids). In an exemplary embodiment, the unsaturated fatty acid is a delta ⁹ unsaturated fatty acids. Delta ⁹ unsaturated fatty acids, position the carbon between the C9 and C10 in the alkene chain of the unsaturated fatty acids - carbon double bond. In determining this position, the alkene chain is numbered starting with the carbon atom in the carbonyl group of the unsaturated fatty acid. In a preferred embodiment, the Δ ⁹ unsaturated starting material has a straight chain alkene chain. Suitable examples of Δ ⁹ unsaturated fatty acids include myristoleic acid, H ₃ C (CH ₂ ) ₃ —CH═CH (CH ₂ ) ₇ COOH, palmitoleic acid, H ₃ C (CH ₂ ) ₅ CH═CH ( CH ₂ ) ₇ COOH, elaidic acid, H ₃ C (CH ₂ ) ₇ CH═CH (CH ₂ ) ₇ COOH and oleic acid, H ₃ C (CH ₂ ) ₇ CH═CH (CH ₂ ) ₇ COOH but not limited to, each of these has a C9-C10 unsaturated bonds (delta ⁹ olefins). In many embodiments, useful delta ⁹ unsaturated fatty acids are obtained from natural oils (e.g., vegetable-based oils or animal fats). Representative examples of renewable plant-based oils include olive oil, peanut oil, grape seed oil, sea buckthorn oil, and sesame oil, poppy seed oil, nutmeg butter, palm oil, coconut oil, macadamia oil, Sea buckthorn oil. Representative examples of animal fats include lard fat and tallow fat. Unsaturated fatty acids can be obtained commercially or synthesized by saponification of fatty acid esters using methods known to those skilled in the art.

  In a preferred embodiment, dodecenedioic acid or a derivative thereof (eg, (Z) -dimethyldodecenedioic acid) is synthesized by a cross metathesis reaction using a metathesis catalyst, as shown in FIG.

  In an exemplary embodiment, oleic acid is converted to dimethyl octadecenedioate in a two-step synthesis process, as described in Ngo and Foglia (JAOCS, 1985, 84: 777-784). For example, oleic acid is placed in the presence of a metathesis catalyst to form dimethyl 9-octadecenedioate. The octadecenedioic acid can then be purified using techniques known in the art.

  In some embodiments, the starting material comprises dimethyl hexenedioate and dimethyl octadecenedioate. After cross-metathesis of dimethyl hexenedioate in the presence of dimethyl octadecenedioate, dimethyl dodecenedioate is formed. The dodecenedioic acid can be converted to dodecanedioic acid by hydrogenation to saturate the olefin. In a preferred embodiment, dimethyl dodecenedioate is reduced to form dimethyl dodecanedioate. In a preferred embodiment, dimethyl hexenedioate derived from biological sources is produced by reduction of a muconic acid derivative. The product obtained from the metathesis reaction is a composition containing a dimethyl dodecanedioate compound derived from a biological source.

  The metathesis reaction is performed under conditions suitable to produce the desired metathesis product. In some embodiments, the metathesis reaction can be performed under an inert atmosphere. Preferably, the inert atmosphere is an inert gas that does not interfere with the metathesis catalyst. Examples of inert gases include, but are not limited to, nitrogen, argon, neon, helium, and combinations thereof. In some embodiments, the metathesis reaction is processed under any desired pressure. In some embodiments, the metathesis reaction is performed in an inert solvent that does not interfere with the catalyst. For example, the inert solvent includes, but is not limited to, aromatic hydrocarbons (eg, benzene or toluene), halogenated aromatic hydrocarbons, and aliphatic hydrocarbons (eg, methanol). Solvents may be desirable when the above reactions are not miscible throughout and can both be dissolved in a suitable solvent. Preferably, the solvent is thermally stable and does not decompose at the process temperature. Furthermore, in some other embodiments, the metathesis reaction proceeds in a solvent-free reaction. The metathesis reaction is performed at a temperature selected to form the desired product and reduce the formation of undesirable products. Typically, the temperature is above 0 ° C, above 20 ° C, above 40 ° C, or above 50 ° C. In an exemplary embodiment, the metathesis reaction temperature is from about 20 ° C to about 100 ° C. For example, the process temperature can be about 45 ° C to about 50 ° C. In some embodiments, the amount of catalyst used in the metathesis reaction is selected to form the desired product and reduce the formation of undesirable products. For example, the molar ratio of the diacid to the catalyst can range from 5: 1 to 20: 1 or ˜100: 1, or ˜1,000: 1, or ˜100,000: 1. In some embodiments, the reaction time is about 1 hour, about 2 hours, about 4 hours, about 5 hours, about 10 hours, about 15 hours, or more.

  After cross metathesis, the product is separated from the starting material and from the catalyst. Useful techniques for separating and purifying the desired product from starting materials or other undesired products include distillation, chromatography, fractional recrystallization, liquid / liquid extraction, or any combination thereof. However, it is not limited to these. Preferably, the desired product is purified to a high degree, for example 90% or more. In some embodiments, the conversion can be checked by gas chromatography-mass spectrometry. For example, the filtrate of the above reaction mixture (cross-metathesis of dimethyl hexenedioate and dimethyl octadecenedioate, and subsequent hydrogenation) can be checked by gas chromatography-mass spectrometry, with a standard dimethyl dodecanedioate calibration curve and Can be compared.

  Oleic acid, octadec-9-enedioic acid and the 3-hexenedioic acid are symmetric molecules with respect to the carboxyl or ester end groups, and thus the self-metathesis of the oleic acid. It should be recognized that the reaction, or cross-metathesis of the octadec-9-enedioic acid and the 3-hexenedioic acid, theoretically results in less product formation than when an asymmetric molecule is used. It is. For example, cross-metathesis of 3-hexenedioic acid and octadec-9-enedioic acid theoretically forms only dimethyl dodecenedioate. Whether cis isomers or trans isomers are formed in this type of reaction depends on the configuration of the above-mentioned catalyst as well as the substituents on the carbon-carbon double bonds of the newly formed molecule. It is determined by the orientation of the molecule assumed in the case of positioning.

  When the metathesis catalyst used for the metathesis process promotes double bond transfer, i.e., the metathesis catalyst removes the double bond from its original position in the unsaturated fatty acid. When moved to either a position close to or further away from the group, the metathesis reaction produces a mixture of dicarboxylic acid products. For example, the metathesis catalyst promotes double bond transfer at the center of the octadec-9-enegioate, such as octadec-8-enegioate, octadec-7-enegioate and octadec-6-enegioate (as shown in FIG. 3). To form an unsaturated fatty acid dimethyl ester by-product having various backbone lengths from 7 to 16 carbons upon cross-metathesis with dimethyl hexenedioate. (As shown in FIG. 4). In some embodiments, the reaction conditions are modified to prevent double bond migration. In some embodiments, the ratio of lower olefin to unsaturated fatty acid or derivative thereof is selected to prevent double bond migration. In one embodiment, the dimethyl hexenedioate is added more than dimethyl octadecenedioate. For example, the reactant dimethyl hexenedioate / dimethyl octadecenedioate molar ratio can be 2: 1, 3: 1, 4: 1 or more. In other embodiments, acidic additives are added to inhibit double bond migration and undesirable product formation. Examples of acidic additives include, but are not limited to, benzoic acid and salts, phosphoric acid and salts.

  Aspects of the invention relate to compositions comprising dodecenedioic acid or dodecenedioic acid derivatives, and at least one unsaturated dicarboxylic acid or unsaturated dicarboxylic acid derivative by-product obtained from dodecenedioic acid or dodecenedioic acid derivatives. In some embodiments, the composition comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8 derived from the dodecenedioic acid or dodecenedioic acid derivative. Seeds, containing at least 9 by-products. In some embodiments, the at least one unsaturated dicarboxylic acid or unsaturated dicarboxylic acid derivative byproduct comprises an alkene chain having from about 7 to about 16 carbon atoms. In some embodiments, the composition comprises at least 9 byproducts derived from the dodecenedioic acid or dodecenedioic acid derivative, wherein the byproduct comprises from about 7 to about 16 carbon atoms. Including alkene chain containing. In some embodiments, the dodecenedioic acid derivative is a dodecenedioic acid diester. Preferably, the alkene chain contains a carbon double bond at the C3-C4 position.

  Another aspect of the present invention is the use of 9-octadecenedioic acid or 9-octadecenedioic acid derivative and at least one octadecenedioic acid or octadecenedioic acid derivative secondary derivative derived from 9-octadecenedioic acid or 9-octadecenedioic acid derivative. It relates to a composition comprising the product. In some embodiments, the composition comprises at least one, at least 3, at least 4, at least 5, at least 6, at least 7 derived from 9-octadecenedioic acid or 9-octadecenedioic acid derivatives. , At least 8, at least 9, at least 10, at least 15 and at least 16 by-products. In some embodiments, the octadecenedioic acid or octadecenedioic acid derivative by-product is C1-C2, C2-C3, C3-C4, C4-C5, C5-C6, C6-C7. , C7-C8 position, C8-C9 position, C10-C11 position, C11-C12 position, C12-C13 position, C13-C14 position, C14-C15 position, C15-C16 position, C16-C17 position, or C17-C18 position Contains a carbon double bond at the position.

(Polyamide production)
As used herein, “polyamide” is a polymer comprising amide monomers linked by amide bonds formed by reacting an amine with a carboxylic acid or derivative thereof. Like the polyesters, individual polyamides have many uses, and the polyamides include aramid, nylon, and polyaspartic acid. Aramids (shown below) are aromatic polyamides made by polymerizing terephthalic acid or its derivatives (eg terephthaloyl chloride) and diamines (eg 1,4-phenyldiamine (para-phenylene). Diamine).

  Nylon is a generic name for a family of synthetic thermoplastic polyamides used to make fabrics, instrument strings, ropes, screws and gears, to name a few. Nylon can also be utilized with fillers (e.g., glass filled and polybuden sulfide filled variants). Nylon 6 is the most common commercial grade molded nylon. The numerical suffix specifies the number of carbon atoms provided by the monomer; the diamine comes first and the diacid comes second. For nylon 6,6, the diamine is typically hexamethylene diamine, and the acid is adipic acid. Each of these monomers gives 6 carbon atoms to the polymer chain.

  Another example of a useful nylon is nylon 6,12, which has a glass transition temperature of 46 ° C. and a molecular weight of repeating units of 310.48 g / mol. The structural formula of nylon 6,12 is shown below.

One method for making nylon 6,12 is the polycondensation product of 1,6-hexamethylenediamine H ₂ N— (CH ₂ ) ₆ —NH ₂ and dodecanedioic acid HOOC— (CH ₂ ) ₁₀ —COOH. (See, eg, US Pat. No. 3,903,152, which is incorporated herein by reference). US Pat. No. 3,903,152 provides a process for the production of nylon 6,12 from dodecanedioic acid, in which 20 g of dodecanedioic acid is combined with 20 g of water to 70 ° C. And neutralized with a 50 wt% aqueous hexamethylenediamine solution. The pH was then adjusted to 8 to prepare a 50% nylon salt aqueous solution. Next, the aqueous nylon salt solution was heated from room temperature to 250 ° C. in a salt bath under a nitrogen atmosphere, and subsequently polymerized at 250 ° C. for 5 hours under normal pressure. WO / 2000/009586 provides another method for making nylon 6,12. According to this document, nylon 6,12 is prepared by combining decanedioic acid and 1,6-hexanediamine aqueous solution in water with stirring in an autoclave at 90 ° C. for 30 minutes. As a result, a 55 wt% salt solution is obtained. Water is first raised to 180 ° C. in 10 minutes, half of the water is removed by distillation, then the temperature is raised to 200 ° C., for example, 90% by weight aqueous salt solution As obtained, a certain amount of water is removed by distillation. The reactor is completely closed, the distillation is stopped, the temperature is raised to 227 ° C. and the polymerization starts. The pressure increases slowly due to the water present and the high temperature. The pressure at the end of the polymerization is about 12 × 10 ⁵ Pa. The prepolymerization is performed at a constant temperature for 1/2 hour, after which the contents of the autoclave are flushed with a nitrogen atmosphere. The prepolymer is cooled in a nitrogen atmosphere. The obtained prepolymer granules are sieved so as to obtain a fraction having a diameter between 1 and 2 mm. This fraction is introduced into either a static bed (solid material with a capacity of about 50 g) or a tumble dryer (with a capacity of about 10 liters) and at high temperature (about 25 ° C. below the melting point of the polymer). Postcondensation in a nitrogen / water vapor (75/25 volume% required) atmosphere for 24 hours. The polymer granules were then cooled to room temperature. Many rods and plates were injection molded from the polymer thus prepared.

  Although the invention has been particularly shown and described with reference to specific embodiments, various changes in form and detail may be found from the spirit and scope of the invention as defined by the appended claims. It should be understood by those skilled in the art that this can be done without departing.

Claims (70)

A method for producing dodecanedioic acid, the method comprising:
Reducing muconic acid to hexenedioic acid;
Reacting the hexenedioic acid with an unsaturated fatty acid in a metathesis reaction to produce dodecenedioic acid; and reducing the dodecenedioic acid to dodecanedioic acid;
Including the method. Said unsaturated fatty acids are reacted in a self-metathesis reaction, step for generating a delta ^{9-octadecenoic} acid, and said hexene diacid, is reacted with the delta ^{9-octadecenoic} diacid, the step of generating dodecene diacid The method of claim 1 further comprising. The method of claim 1, comprising providing muconic acid, wherein the muconic acid is cis, trans-muconic acid. The method of claim 1, wherein muconic acid is produced from a renewable carbon source via biocatalytic conversion. Recombinant cells expressing 3-dehydroshikimate dehydratase, protocatechuate decarboxylase and catechol 1,2-dioxygenase in a medium containing the renewable carbon source, The cells are cultured under conditions where they are converted to 3-dehydroshikimate by enzymes in the normal pathway of cellular aromatic amino acid biosynthesis, and the 3-dehydroshikimate is converted to cis, cis-muconic acid by a biocatalyst. The method of claim 4 further comprising a step. further comprising isomerizing cis, cis-muconic acid to cis, trans-muconic acid under reaction conditions wherein substantially all of the cis, cis-muconic acid is isomerized to cis, trans-muconic acid The method according to claim 4. 6. The method according to claim 5, wherein the recombinant cell is a prokaryotic cell or a yeast cell. The prokaryotic cell may belong to the genus Escherichia, Klebsiella, Corynebacterium, Brevibacterium, Arthrobacter, Bacillus, Pseudomonas, Streptomyces, Staphylococcus, or the genus Staphylococcus. The method according to claim 7, wherein the yeast cell belongs to the genus Saccharomyces or Schizosaccharomyces. 6. The method according to claim 5, wherein the culturing further comprises a step of producing a broth containing the recombinant cells and extracellular muconic acid, and further removing the recombinant cells from the broth. The method according to claim 1, wherein the step of reducing the muconic acid to hexenedioic acid comprises reacting the muconic acid with zinc halide. The unsaturated fatty acid is delta ⁹ unsaturated fatty acids The method of claim 1. 13. The method of claim 12, wherein the [Delta] ⁹ unsaturated fatty acid is myristoleic acid, palmitoleic acid, elaidic acid, oleic acid, or a combination thereof. The method of claim 1, comprising using a metathesis catalyst. The method of claim 14, wherein the catalyst is a Grubbs catalyst. The catalyst is benzylidene-bis (tricyclohexylphosphine) dichlororuthenium or benzylidene [1,3-bis (2,4,6-trimethylphenyl) -2-imidazolidinylidene] dichloro (tricyclohexylphosphine) ruthenium, Item 16. The method according to Item 15. The method of claim 1, comprising hydrogenating the dodecenedioic acid to form dodecanedioic acid. The method of claim 1, further comprising forming a polyamide using the dodecanedioic acid. The method of claim 18, wherein the polyamide is nylon 6,12. The method according to claim 18, comprising reacting 1,6-hexamethylenediamine and dodecanedioic acid. The method of claim 1, wherein the hexenedioic acid is an isomer of 3-hexenedioic acid. The method of claim 1, wherein the renewable carbon source is D-glucose. A method for producing dodecanedioic acid, the method comprising:
Reacting hexenedioic acid with an unsaturated fatty acid in a metathesis reaction, thereby producing dodecenedioic acid; and reducing the dodecenedioic acid to dodecanedioic acid;
Including the method. The unsaturated fatty acids are reacted in a self-metathesis reaction, to generate the delta ^{9-octadecenoic} acid, then encompasses said hexenedioic acid, a step of reacting the delta ^{9-octadecenoic} diacid in the metathesis reaction, wherein Item 24. The method according to Item 23. 24. The method of claim 23, wherein the hexenedioic acid is an isomer of 3-hexenedioic acid and the unsaturated fatty acid is a [Delta] ⁹ unsaturated fatty acid. A method for producing dodecanedioic acid, the method comprising:
Providing muconic acid produced from a renewable carbon source via biocatalytic conversion;
Reducing the muconic acid to hexenedioic acid using a zinc halide reagent;
Step and said hexene diacid, and delta ⁹ unsaturated fatty acids are reacted in the metathesis reaction to produce dodecene dioic acid;
Reducing the dodecenedioic acid to dodecanedioic acid; and forming a polyamide using the dodecanedioic acid;
Including the method. The delta ⁹ unsaturated fatty acids are reacted in a self-metathesis reaction, delta ^{9-octadecenoic} step to produce a diacid, and the reacting the hexenedioic acid and the delta ^{9-octadecenoic} acid, to produce dodecene diacid 27. The method of claim 26, further comprising: 27. The method of claim 26, wherein the [Delta] ⁹ unsaturated fatty acid is myristoleic acid, palmitoleic acid, elaidic acid, oleic acid, or a combination thereof. 27. The method of claim 26, comprising using a metathesis catalyst. The catalyst is benzylidene-bis (tricyclohexylphosphine) dichlororuthenium or benzylidene [1,3-bis (2,4,6-trimethylphenyl) -2-imidazolidinylidene] dichloro (tricyclohexylphosphine) ruthenium, Item 30. The method according to Item 29. 27. The method of claim 26, wherein the polyamide is nylon 6,12. Following formula: HOOC- a _{(CH 2)} n compositions comprising dicarboxylic acid derived from a biological source, represented by -COOH, wherein n is an integer of 4 to 22, the composition. 33. The composition of claim 32, wherein the dicarboxylic acid is dodecanedioic acid or a derivative thereof. 35. The method of claim 32, wherein the biological source-derived dicarboxylic acid comprises a ¹⁴ C / ¹² C ratio greater than zero. 33. The composition of claim 32, wherein the biological source-derived dicarboxylic acid comprises a ¹⁴ C / ¹² C ratio of about 1.2 × 10 ⁻¹² . 34. The composition of claim 33, wherein the dodecanedioic acid derivative is dimethyldodecanedioic acid. A composition comprising a biologically derived dicarboxylic acid diester of the formula R ₁ —OOC— (CH ₂ ) _n —COO—R ₂ , wherein R ₁ and R ₂ are each independently hydrogen or aliphatic A composition wherein n is an integer from 4 to 22. 38. The method of claim 37, wherein the dicarboxylic acid diester constitutes a ¹⁴ C / ¹² C ratio greater than zero. 38. The composition of claim 37, wherein the dicarboxylic acid diester comprises a ¹⁴ C / ¹² C ratio of about 1.2 × 10 ⁻¹² . Following _{formula: HOOC- (CH 2) 7 -CH} = CH- (CH 2) 7 including dodecendioic acid derived from a biological source, represented by -COOH, composition. 41. The composition of claim 40, wherein the dodecenedioic acid comprises a ¹⁴ C / ¹² C ratio greater than zero. The dodecene diacid constitute ¹⁴ C ^{/ 12} C ratio of about 1.2 × ^{10 -12,} composition according to claim 40. A composition comprising a biological source-derived dodecenedioic acid diester represented by the following formula: R ₁ —OOC—CH ₂ —CH═CH— (CH ₂ ) ₇ —COO—R ₂ , wherein R ₁ And R ₂ are each independently hydrogen or an aliphatic group. 44. The composition of claim 43, wherein the dodecenedioic acid diester comprises a ¹⁴ C / ¹² C ratio greater than zero. 44. The composition of claim 43, wherein the dodecenedioic acid diester comprises a ¹⁴ C / ¹² C ratio of about 1.2 × 10 ⁻¹² . A composition comprising 3-hexenedioic acid derived from a biological source represented by the following formula: R ₁ —OOC—CH ₂ —CH═CH—CH ₂ —COO—R ₂ or a 3-hexenedioic acid derivative thereof. Wherein R ₁ and R ₂ are each independently hydrogen or an aliphatic group. 47. The composition of claim 46, wherein the 3-hexenedioic acid or its 3-hexenedioic acid derivative comprises a ¹⁴ C / ¹² C ratio of about 1.2 × 10 ⁻¹² . A composition comprising nylon 6,12 from biological sources, wherein the nylon 6,12 comprises a ¹⁴ C / ¹² C ratio greater than zero. The ¹⁴ C ^{/ 12} C ratio greater than 0.9 × ^{10 -12,} composition according to claim 48. Following formula: HOOC- a _{(CH 2)} n compositions comprising dicarboxylic acid derived from a biological source, represented by -COOH, wherein n is an integer of 4-22, dicarboxylic from biological sources The composition wherein the acid comprises up to about one trillionth carbon 14. A composition comprising a dicarboxylic acid diester derived from a biological source of the formula: R ₁ —OOC— (CH ₃ ) _n —COOR ₂ , wherein R ₁ and R ₂ are each independently hydrogen or an aliphatic group Wherein n is an integer from 4 to 22 and the biologically derived dicarboxylic acid comprises up to about 1 / trillion carbon 14. A composition comprising a dodecanedioic acid diester derived from a biological source of the formula: R ₁ —OOC— (CH ₂ ) _n —COO—R ₂ , wherein R 1 and R 2 are each independently hydrogen or aliphatic A composition wherein the biological source-derived dodecanedioic acid diester comprises up to about 1 trillion carbons 14. A composition comprising 3-hexenedioic acid derived from a biological source represented by the following formula: R ₁ —OOC—CH ₂ —CH═CH—CH ₂ —COO—R ₂ or a 3-hexenedioic acid derivative thereof. Where R ₁ and R ₂ are each independently hydrogen or an aliphatic group, and the biogenic 3-hexenedioic acid contains up to about 1 trillion carbon 14 Composition. A composition comprising nylon 6,12 from a biological source, wherein the nylon 6,12 comprises a detectable trace of carbon 14. The following steps:
Reducing muconic acid to hexenedioic acid;
Reacting the hexenedioic acid and an unsaturated fatty acid in a metathesis reaction to produce an unsaturated dicarboxylic acid; and reducing the unsaturated dicarboxylic acid to form a dicarboxylic acid. A dicarboxylic acid,
Here, the dicarboxylic acid is represented by the following formula: HOOC— (CH ₂ ) _n —COOH, where n is an integer of 4 to 22.
Dicarboxylic acid. 56. The dicarboxylic acid of claim 55, wherein the muconic acid is produced from a renewable carbon source via biocatalytic conversion. 56. The dicarboxylic acid of claim 55, wherein the unsaturated fatty acid is a [Delta] ⁹ unsaturated C18 fatty acid and the dicarboxylic acid is dodecanedioic acid. 58. The dodecanedioic acid of claim 57, wherein the dodecanedioic acid comprises up to about 1 trillionth carbon 14. A method for producing a dicarboxylic acid, the method comprising:
Reducing muconic acid to hexenedioic acid;
Reacting the hexenedioic acid with an unsaturated fatty acid in a metathesis reaction to produce dodecenedioic acid; and reducing the unsaturated dicarboxylic acid to form the dicarboxylic acid;
Including the method. A step of reacting the unsaturated fatty acid in a self-metathesis reaction to produce Δ ⁹ octadecenedioic acid, and a step of reacting the hexenedioic acid and the Δ ⁹ octadecenedioic acid to produce dodecenedioic acid. 60. The method of claim 59, comprising. A composition comprising dodecenedioic acid or dodecenedioic acid derivative, and at least one unsaturated dicarboxylic acid or unsaturated dicarboxylic acid derivative byproduct obtained from said dodecenedioic acid or said dodecenedioic acid derivative. 62. The composition of claim 61, comprising at least two unsaturated dicarboxylic acids or unsaturated dicarboxylic acid derivative byproducts. 62. The composition of claim 61, comprising at least nine unsaturated dicarboxylic acids or unsaturated dicarboxylic acid derivative byproducts. 62. The composition of claim 61, wherein the at least one unsaturated dicarboxylic acid or unsaturated dicarboxylic acid derivative byproduct comprises an alkene chain having from about 7 to about 16 carbon atoms. 62. The composition of claim 61, wherein the dodecenedioic acid derivative is a dodecenedioic acid diester. 62. The composition of claim 61, wherein the dodecenedioic acid or dodecenedioic acid derivative comprises up to about 1 trillion carbon 14. 62. The composition of claim 61, wherein the alkene chain comprises a carbon double bond at the C3-C4 position. 9-octadecenedioic acid or 9-octadecenedioic acid derivative, and C1-C2, C2-C3, C3-C4, C4-C5, C5-C6, C6-C7, C7-C8, C8-C9, C10-C11, C11-C12, C12-C13, C13-C14, C14-C15, C15-C16, C16-C17, or C17-C18 containing at least one octadecene diacid or octadecene diacid derivative containing a carbon double bond A composition comprising a product, wherein the at least one by-product is obtained from 9-octadecenedioic acid or a 9-octadecenedioic acid derivative. 69. The composition of claim 68 comprising at least two by-products obtained from 9-octadecenedioic acid or 9-octadecenedioic acid derivatives. 69. The composition of claim 68, wherein the 9-octadecenedioic acid or 9-octadecenedioic acid derivative comprises up to about 1 trillion carbon 14.