公开(公告)号：US08949032B2

公开(公告)日：2015-02-03

申请号：US11188136

申请日：2005-07-21

Applicant: Imandokht Famili ,  Christophe H. Schilling

Inventor： Imandokht Famili ,  Christophe H. Schilling

IPC: G01N33/50 ,  G06F19/12 ,  G06F19/18 ,  G06F19/28

CPC classification number: G06F19/12 ,  G01N33/5008 ,  G01N2500/10 ,  G06F19/18 ,  G06F19/28 ,  G06F19/704

Abstract: The invention provides a computer readable medium or media, having: (a) a first data structure relating a plurality of reactants to a plurality of reactions from a first cell, each of said reactions comprising a reactant identified as a substrate of the reaction, a reactant identified as a product of the reaction and a stoichiometric coefficient relating said substrate and said product; (b) a second data structure relating a plurality of reactants to a plurality of reactions from a second cell, each of said reactions comprising a reactant identified as a substrate of the reaction, a reactant identified as a product of the reaction and a stoichiometric coefficient relating said substrate and said product; (c) a third data structure relating a plurality of intra-system reactants to a plurality of intra-system reactions between said first and second cells, each of said intra-system reactions comprising a reactant identified as a substrate of the reaction, a reactant identified as a product of the reaction and a stoichiometric coefficient relating said substrate and said product; (d) a constraint set for said plurality of reactions for said first, second and third data structures, and (e) commands for determining at least one flux distribution that minimizes or maximizes an objective function when said constraint set is applied to said first and second data structures, wherein said at least one flux distribution is predictive of a physiological function of said first and second cells. The first, second and third data structures also can include a plurality of data structures. Additionally provided is a method for predicting a physiological function of a multicellular organism. The method includes: (a) providing a first data structure relating a plurality of reactants to a plurality of reactions from a first cell, each of said reactions comprising a reactant identified as a substrate of the reaction, a reactant identified as a product of the reaction and a stoichiometric coefficient relating said substrate and said product; (b) providing a second data structure relating a plurality of reactants to a plurality of reactions from a second cell, each of said reactions comprising a reactant identified as a substrate of the reaction, a reactant identified as a product of the reaction and a stoichiometric coefficient relating said substrate and said product; (c) providing a third data structure relating a plurality of intra-system reactants to a plurality of intra-system reactions between said first and second cells, each of said intra-system reactions comprising a reactant identified as a substrate of the reaction, a reactant identified as a product of the reaction and a stoichiometric coefficient relating said substrate and said product; (d) providing a constraint set for said plurality of reactions for said first, second and third data structures; (e) providing an objective function, and (f) determining at least one flux distribution that minimizes or maximizes an objective function when said constraint set is applied to said first and second data structures, wherein said at least one flux distribution is predictive of a physiological function of said first and second cells.

公开(公告)号：US08889399B2

公开(公告)日：2014-11-18

申请号：US13286135

申请日：2011-10-31

Applicant: Mark J. Burk ,  Stephen J. Van Dien ,  Anthony P. Burgard ,  Wei Niu

Inventor： Mark J. Burk ,  Stephen J. Van Dien ,  Anthony P. Burgard ,  Wei Niu

IPC: C12N1/20 ,  C12P7/18 ,  C12P7/42 ,  C12N9/04 ,  C12N9/88 ,  C12N9/00 ,  C12N9/02 ,  C12P7/52 ,  C12P17/04

CPC classification number: C12N15/52 ,  B01D3/002 ,  C12N9/0006 ,  C12N9/0008 ,  C12N9/88 ,  C12N9/93 ,  C12N15/70 ,  C12N15/81 ,  C12P7/18 ,  C12P7/42 ,  C12P7/52 ,  C12P17/04 ,  C12Y101/01061 ,  C12Y102/01076 ,  C12Y401/01071 ,  C12Y602/01004 ,  Y02P20/52

Abstract: The invention provides a non-naturally occurring microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (1,4-BDO) biosynthetic pathways. The pathways include exogenous nucleic acids encoding a) an α-ketoglutarate decarboxylase; b) a 4-hydroxybutanoate dehydrogenase; c) a 4-hydroxybutyryl-CoA:acetyl-CoA transferase or a butyrate kinase and a phosphotransbutyrylase; d) an aldehyde dehydrogenase, and e) an alcohol dehydrogenase, wherein the exogenous nucleic acids are expressed in sufficient amounts to produce 1,4-butanediol (1,4-BDO). Also provide is a method for the production of 1,4-BDO. The method includes culturing the non-naturally occurring microbial organism having 4-HB and 1,4-BDO biosynthetic pathways substantially anaerobic conditions for a sufficient period of time to produce 1,4-BDO.

Abstract translation: 本发明提供了具有4-羟基丁酸（4-HB）和1,4-丁二醇（1,4-BDO）生物合成途径的非天然存在的微生物生物。 途径包括编码a）α-酮戊二酸脱羧酶的外源核酸; b）4-羟基丁酸脱氢酶; c）4-羟基丁酰-CoA：乙酰-CoA转移酶或丁酸酯激酶和磷酸转磷酸化酶; d）醛脱氢酶，和e）醇脱氢酶，其中外源核酸以足够的量表达以产生1,4-丁二醇（1,4-BDO）。 还提供了用于生产1,4-BDO的方法。 该方法包括培养具有4-HB和1,4-BDO生物合成途径的非天然存在的微生物生物合成途径，基本上是厌氧条件足够的时间以产生1,4-BDO。

公开(公告)号：US20140295511A1

公开(公告)日：2014-10-02

申请号：US14068768

申请日：2013-10-31

Applicant: Genomatica, Inc.

Inventor： Anthony P. Burgard ,  Mark J. Burk ,  Robin E. Osterhout ,  Priti Pharkya

IPC: C12P7/18

CPC classification number: C12P7/18 ,  C07C29/80 ,  C12N9/0006 ,  C12N9/0008 ,  C12N9/1029 ,  C12N9/1096 ,  C12N9/88 ,  C12N15/52 ,  C12Y101/01 ,  C12Y101/01157 ,  C12Y102/01 ,  C12Y203/01 ,  C12Y206/01 ,  C12Y401/01 ,  C12Y402/01 ,  C12Y403/01

Abstract: A non-naturally occurring microbial organism includes a microbial organism having a 1,3-butanediol (1,3-BDO) pathway having at least one exogenous nucleic acid encoding a 1,3-BDO pathway enzyme expressed in a sufficient amount to produce 1,3-BDO. The pathway includes an enzyme selected from a 2-amino-4-ketopentanoate (AKP) thiolase, an AKP dehydrogenase, a 2-amino-4-hydroxypentanoate aminotransferase, a 2-amino-4-hydroxypentanoate oxidoreductase (deaminating), a 2-oxo-4-hydroxypentanoate decarboxylase, a 3-hydroxybutyraldehyde reductase, an AKP aminotransferase, an AKP oxidoreductase (deaminating), a 2,4-dioxopentanoate decarboxylase, a 3-oxobutyraldehyde reductase (ketone reducing), a 3-oxobutyraldehyde reductase (aldehyde reducing), a 4-hydroxy-2-butanone reductase, an AKP decarboxylase, a 4-aminobutan-2-one aminotransferase, a 4-aminobutan-2-one oxidoreductase (deaminating), a 4-aminobutan-2-one ammonia-lyase, a butenone hydratase, an AKP ammonia-lyase, an acetylacrylate decarboxylase, an acetoacetyl-CoA reductase (CoA-dependent, aldehyde forming), an acetoacetyl-CoA reductase (CoA-dependent, alcohol forming), an acetoacetyl-CoA reductase (ketone reducing), a 3-hydroxybutyryl-CoA reductase (aldehyde forming), a 3-hydroxybutyryl-CoA reductase (alcohol forming), a 4-hydroxybutyryl-CoA dehydratase, and a crotonase. A method for producing 1,3-BDO, includes culturing such microbial organisms under conditions and for a sufficient period of time to produce 1,3-BDO.

Abstract translation: 非天然存在的微生物生物体包括具有1,3-丁二醇（1,3-BDO）途径的微生物生物，其具有至少一种编码1,3-BDO途径酶的外源核酸，所述外源核酸以足够的量表达，以产生1 ，3-BDO。 该途径包括选自2-氨基-4-酮戊酸酯（AKP）硫解酶，AKP脱氢酶，2-氨基-4-羟基戊酸转氨酶，2-氨基-4-羟基戊酸酯氧化还原酶（脱氨基），2- 4-羟基戊酸脱羧酶，3-羟基丁醛还原酶，AKP氨基转移酶，AKP氧化还原酶（脱氨），2,4-二氧代戊酸脱羧酶，3-氧代丁醛还原酶（还原酮），3-氧代丁醛还原酶 ），4-羟基-2-丁酮还原酶，AKP脱羧酶，4-氨基丁-2-酮氨基转移酶，4-氨基丁-2-酮氧化还原酶（脱氨基），4-氨基丁-2-酮氨裂解酶 乙酰乙酰辅酶A还原酶（CoA依赖性醛形成），乙酰乙酰辅酶A还原酶（CoA依赖性醇形成），乙酰乙酰辅酶A还原酶（酮），乙酰乙酰辅酶A还原酶 还原），3-羟基丁酰辅酶A还原酶（醛形成），3-羟基 Ryl-CoA还原酶（醇形成），4-羟基丁酰辅酶A脱水酶和巴豆酸酶。 一种生产1,3-BDO的方法包括在条件和足够的时间内培养这些微生物生产1,3-BDO。

公开(公告)号：US20140235815A1

公开(公告)日：2014-08-21

申请号：US14059285

申请日：2013-10-21

Applicant: Genomatica, Inc.

Inventor： Anthony P. Burgard ,  Robin E. Osterhout ,  Stephen J. Van Dien ,  Cara Ann Tracewell ,  Priti Pharkya ,  Stefan Andrae

IPC: C12P7/46 ,  C08G63/00 ,  C07C55/10 ,  C12P7/24 ,  C12N15/70 ,  C12N15/81

CPC classification number: C12P7/46 ,  C07C55/10 ,  C07C59/235 ,  C08G63/00 ,  C12N9/0006 ,  C12N9/0028 ,  C12N9/1007 ,  C12N15/70 ,  C12N15/74 ,  C12N15/77 ,  C12N15/81 ,  C12N15/815 ,  C12P7/24 ,  C12Y101/01244 ,  C12Y105/0102

Abstract: Provided herein is a non-naturally occurring microbial organism having a methanol metabolic pathway that can enhance the availability of reducing equivalents in the presence of methanol. Such reducing equivalents can be used to increase the product yield of organic compounds produced by the microbial organism, such as succinate. Also provided herein are methods for using such an organism to produce succinate.

Abstract translation: 本文提供了具有甲醇代谢途径的非天然存在的微生物生物，其可以在甲醇存在下增强还原当量的可用性。 这种还原当量可以用于增加由微生物生物如琥珀酸酯产生的有机化合物的产物产率。 本文还提供了使用这种生物体来生产琥珀酸盐的方法。

公开(公告)号：US08753850B2

公开(公告)日：2014-06-17

申请号：US13549239

申请日：2012-07-13

Applicant: Mark J. Burk ,  Anthony P. Burgard ,  Robin E. Osterhout ,  Priti Pharkya

Inventor： Mark J. Burk ,  Anthony P. Burgard ,  Robin E. Osterhout ,  Priti Pharkya

IPC: C12P13/00 ,  C12P21/04 ,  C12N1/00 ,  C12N1/20 ,  C12N9/00 ,  C12N15/00 ,  C07H21/04

Abstract: The invention provides a non-naturally occurring microbial organism having a 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway. The microbial organism contains at least one exogenous nucleic acid encoding an enzyme in the respective 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway. The invention additionally provides a method for producing 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid. The method can include culturing a 6-aminocaproic acid, caprolactam or hexametheylenediamine producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding a 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid pathway enzyme in a sufficient amount to produce the respective product, under conditions and for a sufficient period of time to produce 6-aminocaproic acid, caprolactam, hexametheylenediamine or levulinic acid.

公开(公告)号：US20140127765A1

公开(公告)日：2014-05-08

申请号：US14053381

申请日：2013-10-14

Applicant: GENOMATICA, INC.

Inventor： Robin E. Osterhout ,  Anthony P. Burgard

IPC: C12P7/64 ,  C12P7/24 ,  C12P7/04

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

Abstract translation: 本发明提供了包含脂肪醇，脂肪醛或脂肪酸途径的非天然存在的微生物，其中微生物有选择地产生具有特定长度的脂肪醇，脂肪醛或脂肪酸。 还提供了具有脂肪醇，脂肪醛或脂肪酸途径的非天然存在的微生物，其中所述微生物进一步包括乙酰辅酶A途径。 在一些方面，本发明的微生物有选择性的基因破坏或酶减弱，其增加脂肪醇，脂肪醛或脂肪酸的产生。 本发明另外提供使用上述微生物生产脂肪醇，脂肪醛或脂肪酸的方法。

公开(公告)号：US08715971B2

公开(公告)日：2014-05-06

申请号：US12878980

申请日：2010-09-09

Applicant: Priti Pharkya ,  Anthony P. Burgard ,  Robin E. Osterhout ,  Mark J. Burk ,  Jun Sun

Inventor： Priti Pharkya ,  Anthony P. Burgard ,  Robin E. Osterhout ,  Mark J. Burk ,  Jun Sun

IPC: C12P7/04

CPC classification number: C12P7/40 ,  C12N15/52 ,  C12P7/04 ,  C12P7/18

Abstract: The invention provides a non-naturally occurring microbial organism having n-propanol and isopropanol pathways, 1,4-butanediol (14-BDO) and isopropanol pathways, 1,3-butanediol (13-BDO) and isopropanol pathways or methylacrylic acid (MAA) and isopropanolpathways. The microbial organism contains at least one exogenous nucleic acid encoding an enzyme in each of the respective n-propanol, 14-BDO, 13-BDO or MAA and isopropanol pathways. The invention additionally provides a method for co-producing n-propanol and isopropanol, 14-BDO and isopropanol, 13-BDO and isopropanol or MAA and isopropanol. The method can include culturing an n-propanol and an isopropanol co-producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding an n-propanol, an isopropanol, a 14-BDO, a 13-BDO and/or a MAA pathway enzyme in a sufficient amount to produce each of the respective products, under conditions and for a sufficient period of time to produce each of the respective products.

Abstract translation: 本发明提供了具有正丙醇和异丙醇途径的非天然存在的微生物生物，1,4-丁二醇（14-BDO）和异丙醇途径，1,3-丁二醇（13-BDO）和异丙醇途径或甲基丙烯酸（MAA ）和异丙醇通道。 微生物生物体含有至少一种外源核酸，其编码在各自的正丙醇，14-BDO，13-BDO或MAA和异丙醇途径中的酶。 本发明另外提供了共同生产正丙醇和异丙醇，14-BDO和异丙醇，13-BDO和异丙醇或MAA和异丙醇的方法。 该方法可以包括培养正丙醇和异丙醇共同生产的微生物，其中微生物生物体表达至少一种编码正丙醇，异丙醇，14-BDO，13-BDO和/或 足够量的MAA途径酶，以在条件下和足够的时间内产生各个产物中的每一种以产生各个产物。

公开(公告)号：US08697421B2

公开(公告)日：2014-04-15

申请号：US13615168

申请日：2012-09-13

Applicant: Mark J. Burk ,  Christophe H. Schilling ,  Anthony P. Burgard ,  John D. Trawick

Inventor： Mark J. Burk ,  Christophe H. Schilling ,  Anthony P. Burgard ,  John D. Trawick

IPC: C12N1/20 ,  C12N1/00 ,  C12P11/00 ,  C12P7/26 ,  C12P5/02 ,  C12P19/32 ,  C07H21/04

CPC classification number: C12P7/18 ,  C12N9/0008 ,  C12N9/0036 ,  C12N9/1007 ,  C12N9/93 ,  C12N15/70 ,  C12P19/32 ,  Y02E50/343

Abstract: The invention provides a non-naturally occurring microbial organism having an acetyl-CoA pathway and the capability of utilizing syngas or syngas and methanol. In one embodiment, the invention provides a non-naturally occurring microorganism, comprising one or more exogenous proteins conferring to the microorganism a pathway to convert CO, CO2 and/or H2 to acetyl-coenzyme A (acetyl-CoA), methyl tetrahydrofolate (methyl-THF) or other desired products, wherein the microorganism lacks the ability to convert CO or CO2 and H2 to acetyl-CoA or methyl-THF in the absence of the one or more exogenous proteins. The invention additionally provides a method for producing acetyl-CoA, for example, by culturing an acetyl-CoA producing microbial organism, where the microbial organism expresses at least one exogenous nucleic acid encoding an acetyl-CoA pathway enzyme or protein in a sufficient amount to produce acetyl-CoA, under conditions and for a sufficient period of time to produce acetyl-CoA.

Abstract translation: 本发明提供具有乙酰-CoA途径的非天然存在的微生物生物体，以及利用合成气或合成气和甲醇的能力。 在一个实施方案中，本发明提供非天然存在的微生物，其包含一种或多种外源蛋白质，赋予微生物将CO，CO 2和/或H 2转化为乙酰辅酶A（乙酰辅酶A），甲基四氢叶酸（甲基 -THF）或其它所需产物，其中微生物缺乏在不存在一种或多种外源蛋白的情况下将CO或CO 2和H 2转化为乙酰辅酶A或甲基-THF的能力。 本发明另外提供了生产乙酰辅酶A的方法，例如通过培养产生乙酰辅酶A的微生物，其中微生物生物体表达至少一种编码乙酰辅酶A途径酶或蛋白质的外源核酸，其量足够 在条件下和足够的时间内产生乙酰辅酶A产生乙酰辅酶A。

公开(公告)号：US08673601B2

公开(公告)日：2014-03-18

申请号：US12018129

申请日：2008-01-22

Applicant: Anthony P. Burgard ,  Stephen J. Van Dien

Inventor： Anthony P. Burgard ,  Stephen J. Van Dien

IPC: C12P7/52 ,  C12N9/00 ,  C12N1/20 ,  C12N15/00 ,  C07H21/04

CPC classification number: C12N15/52 ,  C12N9/00 ,  C12P7/42

Abstract: The invention provides a non-naturally occurring microorganism having one or more gene disruptions, the one or more gene disruptions occurring in genes encoding an enzyme obligatory coupling 3-hydroxypropionic acid production to growth of the microorganism when the gene disruption reduces an activity of the enzyme, whereby the one or more gene disruptions confers stable growth-coupled production of 3-hydroxypropionic acid onto the non naturally occurring microorganism. The disruptions can be complete gene disruptions and the non-naturally occurring organisms can include a variety of prokaryotic or eukaryotic microorganisms. A method of producing a non-naturally occurring microorganism having stable growth-coupled production of 3-hydroxypropionic acid is further provided. The method includes: (a) identifying in silico a set of metabolic modifications requiring 3-hydroxypropionic acid production during exponential growth, and (b) genetically modifying a microorganism to contain the set of metabolic modifications requiring 3-hydroxypropionic acid production.

Abstract translation: 本发明提供了一种具有一个或多个基因破坏的非天然存在的微生物，当基因破坏降低酶的活性时，编码酶强制性偶合3-羟基丙酸产生的微生物生长的基因中发生的一种或多种基因破坏 由此一个或多个基因破坏赋予非天然存在的微生物上稳定的3-羟基丙酸生长偶联产生。 破坏可能是完全的基因破坏，非天然存在的生物体可以包括多种原核或真核微生物。 还提供了一种生产具有稳定的3-羟基丙酸的生长偶联生产的非天然存在的微生物的方法。 该方法包括：（a）在指数生长过程中，以计算机识别需要3-羟基丙酸生成的一组代谢修饰，和（b）遗传修饰微生物以包含需要3-羟基丙酸生产的一组代谢修饰。

公开(公告)号：US08663957B2

公开(公告)日：2014-03-04

申请号：US12780802

申请日：2010-05-14

Applicant: Robin E. Osterhout ,  Anthony P. Burgard ,  Mark J. Burk ,  Priti Pharkya

Inventor： Robin E. Osterhout ,  Anthony P. Burgard ,  Mark J. Burk ,  Priti Pharkya

IPC: C12P7/26

CPC classification number: C12P7/26 ,  C12N9/16 ,  C12N9/88 ,  C12N9/93 ,  C12N15/52 ,  C12N15/70

Abstract: A non-naturally occurring microbial organism has cyclohexanone pathways that include at least one exogenous nucleic acid encoding a cyclohexanone pathway enzyme. A pathway includes a 2-ketocyclohexane-1-carboxyl-CoA hydrolase (acting on C—C bond), a 2-ketocyclohexane-1-carboxylate decarboxylase and an enzyme selected from a 2-ketocyclohexane-1-carboxyl-CoA hydrolase (acting on thioester), a 2-ketocyclohexane-1-carboxyl-CoA transferase, and a 2-ketocyclohexane-1-carboxyl-CoA synthetase. A pathway includes an enzyme selected from a 6-ketocyclohex-1-ene-1-carboxyl-CoA hydrolase (acting on C—C bond), a 6-ketocyclohex-1-ene-1-carboxyl-CoA synthetase, a 6-ketocyclohex-1-ene-1-carboxyl-CoA hydrolase (acting on thioester), a 6-ketocyclohex-1-ene-1-carboxyl-CoA transferase, a 6-ketocyclohex-1-ene-1-carboxyl-CoA reductase, a 6-ketocyclohex-1-ene-1-carboxylate decarboxylase, a 6-ketocyclohex-1-ene-1-carboxylate reductase, a 2-ketocyclohexane-1-carboxyl-CoA synthetase, a 2-ketocyclohexane-1-carboxyl-CoA transferase, a 2-ketocyclohexane-1-carboxyl-CoA hydrolase (acting on thioester), a 2-ketocyclohexane-1-carboxylate decarboxylase, and a cyclohexanone dehydrogenase. A pathway includes an adipate semialdehyde dehydratase, a cyclohexane-1,2-diol dehydrogenase, and a cyclohexane-1,2-diol dehydratase. A pathway includes a 3-oxopimelate decarboxylase, a 4-acetylbutyrate dehydratase, a 3-hydroxycyclohexanone dehydrogenase, a 2-cyclohexenone hydratase, a cyclohexanone dehydrogenase and an enzyme selected from a 3-oxopimeloyl-CoA synthetase, a 3-oxopimeloyl-CoA hydrolase (acting on thioester), and a 3-oxopimeloyl-coA transferase. Each these pathways can include a PEP carboxykinase. A method for producing cyclohexanone includes culturing these non-naturally occurring microbial organisms.

Abstract translation: 非天然存在的微生物有机体具有包含至少一种编码环己酮途径酶的外源核酸的环己酮途径。 途径包括2-酮环己烷-1-羧基-CoA水解酶（作用于CC键），2-酮环己烷-1-羧酸脱羧酶和选自2-酮环己烷-1-羧基-CoA水解酶的酶（作用于硫酯 ），2-酮环己烷-1-羧基-CoA转移酶和2-酮环己烷-1-羧基-CoA合成酶。 途径包括选自6-酮环己-1-烯-1-羧基-CoA水解酶（作用于CC键）的酶，6-酮环己-1-烯-1-羧基-CoA合成酶，6-酮环己基 - 1-烯-1-羧基-CoA水解酶（作用于硫酯），6-酮环己-1-烯-1-羧基-CoA转移酶，6-酮环己-1-烯-1-羧基-CoA还原酶，6 6-酮环己-1-烯-1-羧酸脱羧酶，6-酮环己-1-烯-1-羧酸还原酶，2-酮环己烷-1-羧基-CoA合成酶，2-酮环己烷-1-羧基-CoA转移酶， 2-酮环己烷-1-羧基-CoA水解酶（作用于硫酯），2-酮环己烷-1-羧酸脱羧酶和环己酮脱氢酶。 途径包括己二酸半醛脱水酶，环己烷-1,2-二醇脱氢酶和环己烷-1,2-二醇脱水酶。 途径包括3-氧代邻苯二甲酸脱羧酶，4-乙酰基丁酸脱水酶，3-羟基环己酮脱氢酶，2-环己烯酮水合酶，环己酮脱氢酶和选自3-氧代木糖酰辅酶A合成酶，3-氧代木糖酰-CoA水解酶 （作用于硫酯）和3-氧代木糖酰辅酶A转移酶。 每个这些途径可以包括PEP羧基激酶。 生产环己酮的方法包括培养这些非天然存在的微生物。