[go: up one dir, main page]

WO2025159558A1 - Micro-organisme recombiné présentant une capacité accrue de production d'acide 3-hydroxypropionique - Google Patents

Micro-organisme recombiné présentant une capacité accrue de production d'acide 3-hydroxypropionique

Info

Publication number
WO2025159558A1
WO2025159558A1 PCT/KR2025/001440 KR2025001440W WO2025159558A1 WO 2025159558 A1 WO2025159558 A1 WO 2025159558A1 KR 2025001440 W KR2025001440 W KR 2025001440W WO 2025159558 A1 WO2025159558 A1 WO 2025159558A1
Authority
WO
WIPO (PCT)
Prior art keywords
hydroxypropionic acid
puuc
ydcw
trc
recombinant microorganism
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/KR2025/001440
Other languages
English (en)
Korean (ko)
Inventor
송찬우
권민아
박종명
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
GS Caltex Corp
Original Assignee
GS Caltex Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by GS Caltex Corp filed Critical GS Caltex Corp
Publication of WO2025159558A1 publication Critical patent/WO2025159558A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0008—Oxidoreductases (1.) acting on the aldehyde or oxo group of donors (1.2)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/93—Ligases (6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/52—Propionic acid; Butyric acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y102/00—Oxidoreductases acting on the aldehyde or oxo group of donors (1.2)
    • C12Y102/01—Oxidoreductases acting on the aldehyde or oxo group of donors (1.2) with NAD+ or NADP+ as acceptor (1.2.1)
    • C12Y102/01003—Aldehyde dehydrogenase (NAD+) (1.2.1.3)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y602/00—Ligases forming carbon-sulfur bonds (6.2)
    • C12Y602/01—Acid-Thiol Ligases (6.2.1)
    • C12Y602/01001—Acetate-CoA ligase (6.2.1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/22—Klebsiella

Definitions

  • the present invention relates to a recombinant microorganism with enhanced 3-hydroxypropionic acid production ability.
  • 3-Hydroxypropionic acid (3-HP) can be produced through chemical synthesis and microbial fermentation processes. Recently, research has been conducted on methods for producing 3-HP using bacteria such as Escherichia coli and Klebsiella.
  • Korean Patent Publication No. 10-2020-0051375 discloses a technology for producing 3-HP using a 3-HP-producing microorganism transformed with a gene encoding adenosyltransferase.
  • the present inventors while studying a recombinant microorganism with high 3-hydroxypropionic acid production ability, completed the present invention by producing a recombinant microorganism with low by-product production and high 3-hydroxypropionic acid production ability.
  • the purpose of the present invention is to provide a recombinant microorganism having low by-product production and high 3-hydroxypropionic acid production capacity.
  • the present invention provides a recombinant microorganism having enhanced 3-hydroxypropionic acid production ability, wherein the pathway for converting 3-hydroxypropionaldehyde into 3-hydroxypropionic acid and the pathway for converting acetate into acetyl CoA are enhanced in a microorganism having a 3-hydroxypropionic acid biosynthetic pathway.
  • the present invention also provides a method for producing 3-hydroxypropionic acid using the recombinant microorganism.
  • the recombinant microorganism of the present invention has the advantage of not only having a high 3-hydroxypropionic acid production ability but also having a low by-product production ability, making it easy to produce and recover 3-hydroxypropionic acid.
  • Figure 1 shows changes in metabolic pathways through gene overexpression in a recombinant microorganism of the present invention. It shows a process of simultaneously overexpressing the puuC gene and the ydcW gene to promote the pathway of converting 3-hydroxypropionaldehyde to 3-hydroxypropionic acid, and a process of overexpressing the acs gene to promote the pathway of converting acetate to acetyl CoA.
  • the trc-puuC and trc-ydcW genes are inserted into the positions of the major byproduct-producing genes dhaT, yqhD, ldhA, and glpK to increase the copy numbers of the puuC and ydcW genes.
  • Figure 2 shows the pathway for producing 3-hydroxypropionic acid from glycerol and the results of analysis of the production amount and yield of 3-hydroxypropionic acid according to the location on the genome for overexpressing the puuC gene and the ydcW gene and the type of each promoter in the recombinant microorganism of the present invention.
  • Figure 3 shows the results of analysis of the production amount and yield of 3-hydroxypropionic acid according to the increase in the number of copies of the puuC gene and the ydcW gene in the genome of the recombinant microorganism of the present invention.
  • Figure 4 shows the results of analyzing the amount and yield of 3-hydroxypropionic acid and the amount of acetate produced according to the location and promoter on the genome for overexpressing the acs gene in the recombinant microorganism of the present invention.
  • This relates to a recombinant microorganism with enhanced 3-hydroxypropionic acid production capacity and an enhanced pathway.
  • This relates to a recombinant microorganism with enhanced 3-hydroxypropionic acid production ability, which has an enhanced pathway for converting acetate to acetyl CoA.
  • This relates to a recombinant microorganism with enhanced 3-hydroxypropionic acid production ability, characterized in that the genes puuC and ydcW encoding aldehyde dehydrogenase involved in the pathway for converting to 3-hydroxypropionic acid and the gene acs encoding acetyl CoA synthetase involved in the pathway for converting to acetyl CoA are each overexpressed.
  • a gene encoding an enzyme having at least 90% amino acid sequence homology with the expression product of the gene puuC (SEQ ID NO: 1) or ydcW (SEQ ID NO: 2) encoding the aldehyde dehydrogenase may also be preferably used, and furthermore, a gene encoding an enzyme having at least 90% amino acid sequence homology with the expression product of the gene acs (SEQ ID NO: 47) encoding the acetyl CoA synthetase may also be preferably used.
  • puuC, ydcW and acs genes of the present invention may be characterized in that they are derived from Klebsiella pneumoniae, but are not limited to those derived from a specific microorganism, and it will be apparent that any gene derived from a microorganism with a similar function to these may be used in the present invention.
  • This relates to a recombinant microorganism with enhanced 3-hydroxypropionic acid production ability, characterized in that the genes of puuC and ydcW are simultaneously overexpressed within the genome and the number of copies of the simultaneous overexpression is arranged in a repeating manner of 1 to 20 times.
  • This relates to a recombinant microorganism with enhanced 3-hydroxypropionic acid production ability, characterized in that the puuC, ydcW and acs genes are each operably linked to a trc promoter.
  • a step of preparing a culture solution by culturing a medium inoculated with the recombinant microorganism
  • a method for producing 3-hydroxypropionic acid comprising a step of recovering 3-hydroxypropionic acid from the above culture solution.
  • the main metabolic pathways of the wild type strain were confirmed, and among them, the puuC gene and the ydcW gene, which are key genes for the production of 3-hydroxypropionic acid, were confirmed, which are genes encoding aldehyde dehydrogenase involved in the conversion of 3-hydroxypropionaldehyde to 3-hydroxypropionic acid.
  • the puuC gene utilizes NAD+ with high efficiency
  • the ydcW gene utilizes NADP+ with high efficiency.
  • the acs gene which converts acetate into acetyl CoA, was identified, and when it was overexpressed, it was found that the production of acetate, a byproduct, was reduced and the production of 3-hydroxypropionic acid was further increased, thereby completing the present invention.
  • the microorganism of the present invention has a 3-hydroxypropionic acid biosynthetic pathway.
  • the microorganism may be a microorganism having the aforementioned 3-hydroxypropionic acid biosynthetic pathway, and is not particularly limited thereto.
  • the 3-hydroxypropionic acid biosynthetic pathway refers to a pathway by which 3-hydroxypropionic acid is synthesized from a specific metabolite in the microorganism.
  • the 3-hydroxypropionic acid biosynthetic pathway of the present invention may be a pathway by which 3-hydroxypropionic acid (3-HP) is synthesized from 3-hydroxypropionaldehyde (3-HPA).
  • the microorganism of the present invention has a pathway by which 3-hydroxypropionic acid is synthesized from a carbon source such as glycerol or glycerin.
  • the microorganism of the present invention may be a microorganism having a 3-hydroxypropionic acid biosynthetic pathway as a wild type or a recombinant microorganism obtained by genetic recombination.
  • the microorganism may be selected from the group consisting of the genus Klebsiella, the genus Enterobacter, the genus Corynebacterium, the genus Clostridium, and the genus Lactobacillus, preferably the genus Klebsiella, and more preferably the genus Klebsiella, and more preferably the genus Klebsiella pneumoniae .
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid in which the pathway for converting 3-hydroxypropionaldehyde into 3-hydroxypropionic acid is enhanced.
  • the pathway for converting 3-hydroxypropionaldehyde into 3-hydroxypropionic acid may be a pathway in which the conversion of 3-hydroxypropionaldehyde into 3-hydroxypropionic acid is controlled by aldehyde dehydrogenase.
  • Aldehyde dehydrogenase regulates the conversion of 3-hydroxypropionaldehyde to 3-hydroxypropionic acid.
  • the pathway for converting 3-hydroxypropionaldehyde to 3-hydroxypropionic acid can be promoted.
  • the increase in aldehyde dehydrogenase activity can be achieved by amplifying the expression of the aldehyde dehydrogenase gene, increasing the enzymatic activity of aldehyde dehydrogenase, etc.
  • a person skilled in the art can increase the activity of aldehyde dehydrogenase by selecting an appropriate method, such as introducing puuC or ydcW, a gene encoding aldehyde dehydrogenase, into a microorganism, causing a mutation in the gene (mutation such as changing, substituting, or deleting some bases or introducing some bases to amplify the expression of the gene), regulating gene expression during the transcription or translation process, replacing or modifying a promoter, etc.
  • an appropriate method such as introducing puuC or ydcW, a gene encoding aldehyde dehydrogenase, into a microorganism, causing a mutation in the gene (mutation such as changing, substituting, or deleting some bases or introducing some bases to amplify the expression of the gene), regulating gene expression during the transcription or translation process, replacing or modifying a promoter, etc.
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid with an enhanced pathway for converting acetate to acetyl CoA.
  • the pathway for converting acetate to acetyl CoA may be a pathway in which the conversion of acetate to acetyl CoA is regulated by acetyl CoA synthetase.
  • Acetyl CoA synthase regulates the conversion of acetate into acetyl CoA.
  • the pathway for converting acetate into acetyl CoA can be promoted.
  • the increase in the activity of the acetyl CoA synthase can be achieved by amplifying the expression of the acetyl CoA synthase gene, increasing the enzymatic activity of the acetyl CoA synthase, etc.
  • a person skilled in the art can increase the activity of the acetyl CoA synthase by selecting an appropriate method, such as introducing the gene acs encoding the acetyl CoA synthase into a microorganism, causing a mutation in the gene (mutation such as changing, substituting, or deleting some bases, or introducing some bases to amplify the expression of the gene), regulating gene expression during the transcription or translation process, replacing or modifying the promoter, etc.
  • an appropriate method such as introducing the gene acs encoding the acetyl CoA synthase into a microorganism, causing a mutation in the gene (mutation such as changing, substituting, or deleting some bases, or introducing some bases to amplify the expression of the gene), regulating gene expression during the transcription or translation process, replacing or modifying the promoter, etc.
  • the recombinant microorganism with enhanced 3-hydroxypropionic acid (3-HP) production ability of the present invention may be a recombinant microorganism with enhanced 3-hydroxypropionic acid production ability characterized by enhanced pathways for converting 3-hydroxypropionaldehyde (3-HPA) into 3-hydroxypropionic acid (3-HP) and for converting acetate into acetyl CoA.
  • the genes that promote the pathway for converting 3-HPA into 3-HP are puuC and ydcW, and the gene that promotes the pathway for converting acetate into acetyl CoA is acs, as described above.
  • the genes that increase the productivity of 3-HP refer to the three genes puuC, ydcW, and acs. Hereinafter, these three genes may also be referred to as 3-HP productivity enhancing genes.
  • the number of copies of the 3-hydroxypropionate (3-HP) production enhancing gene inserted into the genomic DNA is preferably 1 copy or more, more preferably 2 copies or more of the combination of ydcW and puuC, and most preferably 3 copies or more and 20 copies or less.
  • the 3-HP production enhancing genes being replicated are puuC and ydcW, and the acs gene may be inserted alone at the end of a certain copy number, but the location of the inserted acs gene may be anywhere in the genome.
  • aldehyde dehydrogenase is an enzyme that converts 3-hydroxypropionaldehyde into 3-hydroxypropionic acid, and the enzyme may be encoded by the puuC gene or encoded by the ydcW gene, but is not limited thereto.
  • the above puuC gene or ydcW gene may be derived from Klebsiella pneumoniae and may include a mutation in the gene and/or amino acid sequence within the range of maintaining the enzymatic activity that converts 3-HPA into 3-HP.
  • acetyl CoA synthase is an enzyme that converts acetate into acetyl CoA, and the enzyme may be encoded by the acs gene and may be derived from Klebsiella pneumoniae.
  • the recombinant microorganism of the present invention may additionally comprise an inducible promoter and a 3-HP production enhancing gene operably linked thereto, i.e., a puuC gene, a ydcW gene, and/or an acs gene.
  • the inducible promoter may be one that induces expression of a gene by linking to a promoter through the addition (treatment) of an inducer, and the transcription timing of the genes may be controlled using the inducible promoter.
  • the inducible promoter may be BAD, LTTR, trc, and/or tac
  • the microorganism may additionally comprise a puuC gene, a ydcW gene, and/or an acs gene operably linked to a trc promoter.
  • the trc promoter and the genes may be cloned into a single vector and introduced into a cell.
  • the genes may use not only an inducible promoter but also a constitutive promoter for the purpose of increasing the expression amount.
  • promoter refers to an untranslated nucleic acid sequence upstream of a coding region, which includes a binding site for a polymerase and has transcription initiation activity into mRNA of a gene downstream of the promoter, i.e., a DNA region to which a polymerase binds to initiate transcription of a gene, and may be located at the 5' site of the mRNA transcription initiation site.
  • a method in order to increase the productivity of 3-hydroxypropionic acid, a method is used in which the copy number of a gene (polynucleotide) encoding aldehyde dehydrogenase is increased and integrated into the genome, and a gene encoding acetyl CoA synthetase is also inserted, and each gene is operably linked to a trc promoter.
  • the recombinant microorganism with enhanced 3-hydroxypropionic acid production ability of the present invention may have higher productivity (amount of 3-hydroxypropionic acid produced per unit time, per unit volume), productivity (amount of 3-hydroxypropionic acid produced per unit volume, i.e., concentration of 3-hydroxypropionic acid in the culture medium), and yield (production amount of 3-hydroxypropionic acid relative to the amount of carbon source) of 3-hydroxypropionic acid, and in particular, the recombinant microorganism may have a higher concentration of 3-hydroxypropionic acid in the fermentation medium than a wild-type microorganism during fermentation.
  • productivity is defined and used as a broad concept that includes yield in addition to the general concentration concept of productivity.
  • the amount of acetate produced may be lower than that of a wild-type microorganism.
  • the recombinant microorganism of the present invention integrated the puuC gene and the ydcW gene, or the ydcW gene and the puuC gene, which are involved in encoding the enzyme for increasing the activity of aldehyde dehydrogenase, which converts 3-hydroxypropionaldehyde to 3-hydroxypropionic acid, into the genome so that they were simultaneously expressed regardless of the order, thereby confirming that the genome stability was also increased.
  • both puuC and ydcW genes can utilize NAD+ and NADP+ as cofactors, but puuC is known to utilize NAD+ with high efficiency (high cofactor preference) and ydcW is known to utilize NADP+ with high efficiency.
  • puuC and ydcW for simultaneous expression is most desirable in terms of the production amount and yield of 3-hydroxypropionic acid, and in addition to inducible promoters such as trc and tac, various constitutive promoters can be used for the expression of the two genes.
  • the acs gene involved in encoding acetyl CoA synthetase which converts acetate into acetyl CoA, was overexpressed to increase the activity of this enzyme.
  • the strain with an enhanced 3-HP production pathway was designed to express the trc promoter and acs gene, and as a result, it was found that the 3-HP production ability was the best.
  • the present invention relates to a method for producing 3-hydroxypropionic acid, comprising the steps of: inoculating a medium with a recombinant microorganism for producing 3-hydroxypropionic acid of the present invention; culturing the medium inoculated with the recombinant microorganism to produce a culture solution; and recovering 3-hydroxypropionic acid from the culture solution.
  • the above cultivation may be performed as a flask culture without DO (dissolved oxygen) control, or may be performed under aerobic conditions, preferably microaerobic conditions.
  • the above cultivation may be performed while supplying oxygen, i.e. air, during the cultivation, and as a specific example, this may be accomplished through stirring, but is not limited thereto.
  • the recombinant microorganism of the present invention may be cultured in a complex medium, and the type of the complex medium is not particularly limited, and it is obvious that a person skilled in the art can appropriately select and use a commercially available, commonly used complex medium.
  • the culture may be cultured under conditions that do not control DO, or may be performed under conditions of DO of 0 to 10%, 0 to 10%, 1 to 8%, 2 to 7%, 2.5 to 6.5%, or 5%.
  • the culturing can be performed for 1 hour or more, 2 hours or more, 3 hours or more, 4 hours or more, 5 hours or more, 6 hours or more, 7 hours or more, 8 hours or more, 9 hours or more, 10 hours or more, 11 hours or more, 12 hours or more, 13 hours or more, 14 hours or more, 15 hours or more, 16 hours or more, 20 hours or more, 25 hours or more, 30 hours or more, 35 hours or more, 40 hours or more, 45 hours or more, or 48 hours or more.
  • the culturing can be performed for 95 hours or less, 90 hours or less, 85 hours or less, 80 hours or less, 70 hours or less, 65 hours or less, 60 hours or less, 55 hours or less, 50 hours or less, 45 hours or less, 40 hours or less, 35 hours or less, 30 hours or less, 25 hours or less, 20 hours or less, or 15 hours or less.
  • the culture can be performed in a flask for 1 to 24 hours under conditions of initial pH 7.00, 200 rpm, and 37°C.
  • the data shown in Figures 2 to 4 are the results of culture under these conditions.
  • the step of recovering 3-hydroxypropionic acid from the above culture solution is not particularly limited and may be performed by a conventional method for recovering a specific product from a culture solution using microorganisms.
  • the above method for producing 3-hydroxypropionic acid may additionally include a step of purifying the culture solution.
  • the purification may be performed using a method commonly used for purifying 3-hydroxypropionic acid, and is not particularly limited.
  • the carbon source during the above cultivation can be glycerol, glycerin, etc.
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid in which the pathway for converting 3-hydroxypropionaldehyde into 3-hydroxypropionic acid is enhanced among microorganisms having a 3-hydroxypropionic acid biosynthetic pathway.
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid with an enhanced pathway for converting acetate into acetyl CoA.
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid with increased 3-hydroxypropionic acid production ability.
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid in which the pathway for converting 3-hydroxypropionaldehyde into 3-hydroxypropionic acid is promoted and/or the pathway for converting acetate into acetyl CoA is promoted in a microorganism having a 3-hydroxypropionic acid biosynthetic pathway (see FIG. 1).
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid, which has a 3-hydroxypropionic acid concentration in a culture medium of 1 g/L or more, 2 g/L or more, 3 g/L or more, 4 g/L or more, 5 g/L or more, 6 g/L or more, 7 g/L or more, 8 g/L or more, 9 g/L or more, 10 g/L or more, 11 g/L or more, 12 g/L or more, 13 g/L or more, 14 g/L or more, or 15 g/L or more when cultured in a flask for 24 hours under conditions of initial pH 7.00, 200 rpm, and 37°C.
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid, which has a 3-hydroxypropionic acid concentration in a culture medium of 30 g/L or less, 29 g/L or less, 28 g/L or less, 27 g/L or less, 26 g/L or less, 25 g/L or less, 24 g/L or less, 23 g/L or less, 22 g/L or less, 21 g/L or less, 20 g/L or less, 19 g/L or less, 18 g/L or less, 17 g/L or less, or 16 g/L or less when cultured in a flask for 24 hours under initial pH 7.00, 200 rpm, and 37°C conditions.
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid, which has an acetate concentration in a culture medium of 15 g/L or less, 10 g/L or less, 5 g/L or less, 4.5 g/L or less, 4.0 g/L or less, 3.5 g/L or less, 3.0 g/L or less, 2.5 g/L or less, 2.0 g/L or less, 1.5 g/L or less, or 1.0 g/L or less when cultured in a flask for 24 hours under conditions of initial pH 7.00, 200 rpm, and 37°C.
  • the recombinant microorganism of the present invention may be a recombinant microorganism for producing 3-hydroxypropionic acid, which has an acetate concentration in a culture medium of 0.1 g/L or more, 0.15 g/L or more, 0.2 g/L or more, 0.25 g/L or more, 0.3 g/L or more, 0.35 g/L or more, 0.4 g/L or more, 0.45 g/L or more, 0.5 g/L or more, 0.55 g/L or more, 0.6 g/L or more, 0.65 g/L or more, 0.7 g/L or more, or 0.75 g/L or more when cultured in a flask for 24 hours under conditions of initial pH 7.00, 200 rpm, and 37°C.
  • Klebsiella pneumoniae GSC123 (KCTC12133BP) was used.
  • the metabolic product production capacity of microorganisms was calculated as follows.
  • (1) medium, (2) culture conditions, and (3) operating conditions are as follows.
  • Seed cultures are prepared in test tubes and are as shown in Table 1 below.
  • the culture conditions for the main culture are as shown in Tables 2 and 3.
  • the operating conditions of this culture are as shown in Table 4.
  • the operating conditions refer to the basic conditions of flask culture.
  • genes encoding aldehyde dehydrogenase were amplified from the genome of Klebsiella pneumoniae GSC123 strain. Specifically, the expression levels of the puuC and ydcW genes present in the genome were enhanced by expressing them through the strong trc promoter. The trc-puuC and trc-ydcW fragments were obtained through overlapping PCR, and the fragments were fused with the homologous region at the desired insertion site to obtain the final integration fragment.
  • the trc-puuC or trc-ydcW fragment was inserted into the puuC or ydcW site in the genome to produce puuC::trc-puuC, puuC::trc-ydcW, ydcW::trc-puuC, ydcW::trc-ydcW strains, thereby confirming the effect of simultaneous expression of the puuC and ydcW genes (Examples 1 to 8).
  • puuC and ydcW the major byproduct-producing genes identified in the previously applied patent (KR Application No.: 10-2023-0017522), such as dhaT, yqhD, ldhA, glpK, pta-ack, and poxB, were utilized (reference: Fig. 1).
  • a strong promoter such as trc or tac
  • Example 1 Construction of puuC::trc-puuC strain, ydcW::trc-ydcW strain, and puuC::trc-puuC ydcW::trc-ydcW strain
  • the left homologous region was obtained using the primer of SEQ ID NO. (3/4), and the right homologous region was obtained using the primer of SEQ ID NO. (5/6).
  • An overlapping fragment was obtained using the primer of SEQ ID NO. (3/6), and the overlapping fragment was treated with the restriction enzyme (Xba1/Not1) and then introduced into the pKOV plasmid to obtain pKOV-puuC::trc-puuC.
  • the pKOV-puuC::trc-puuC plasmid was introduced into the strain using electroporation (electroporation, 25 uF, 200 ⁇ , 2.5 kV/cm), and finally, the puuC gene in the genome was expressed under the trc promoter (Example 1-1).
  • the ydcW::trc-ydcW strain was also produced using the same method (using SEQ ID NOs. 7/8 and 9/10) (Example 1-2).
  • puuC::trc-puuC ydcW::trc-ydcW strain was also produced using pKOV-puuC::trc-puuC and pKOV-ydcW::trc-ydcW (Example 1-3).
  • the trc-puuC or trc-ydcW region was amplified by PCR in the genome and inserted into the desired insertion site through homologous recombination.
  • trc-puuC was inserted into the ydcW site present in the genome, and trc-puuC was amplified as SEQ ID NO: 11/12, the left homologous arm was amplified with the SEQ ID NO: 13/14 primer, and the right homologous arm was amplified with the SEQ ID NO: 15/16 primer.
  • a homologous recombination site was produced through overlapping PCR using the SEQ ID NO: 11/16 primer. The produced site was introduced into the pKOV plasmid and used for strain production.
  • the overlapping fragment was introduced into the pKOV plasmid after restriction enzyme treatment with (Xba1/Xma1), thereby obtaining pKOV-ydcW::trc-puuC, and this was introduced into the puuC::trc-puuC strain produced in Example 1-1 using the electroporation method described above, thereby producing the GSC123 puuC::trc-puuC ydcW::trc-puuC strain.
  • Example 3 Construction of ydcW::trc-ydcW puuC::trc-ydcW strain
  • trc-ydcW was inserted into the puuC site in the genome, and trc-ydcW was amplified as SEQ ID NO: 17/18, the left homologous arm was amplified with the SEQ ID NO: 19/20 primer, and the right homologous arm was amplified with the SEQ ID NO: 21/22 primer.
  • a homologous recombination site was created through overlapping PCR using the SEQ ID NO: 17/22 primer. The created site was introduced into the pKOV plasmid and used for strain creation.
  • the overlapping fragment was introduced into the pKOV plasmid after restriction enzyme treatment with (Xba1/Xma1), thereby obtaining pKOV-puuC::trc-ydcW, and this was introduced into the ydcW::trc-ydcW strain produced in Example 1-2 using the electroporation method described above, thereby producing the GSC123 ydcW::trc-ydcW puuC::trc-ydcW strain.
  • trc-puuC was inserted into the dhaT site of the byproduct-producing gene present in the genome, and trc-puuC was amplified as SEQ ID NO: 23/24, the left homologous arm was amplified with the SEQ ID NO: 25/26 primer, and the right homologous arm was amplified with the SEQ ID NO: 27/28 primer.
  • a homologous recombination site was produced through overlapping PCR using the SEQ ID NO: 23/28 primer. The produced site was introduced into the pKOV plasmid and used for strain production.
  • the overlapping fragment was introduced into the pKOV plasmid after restriction enzyme treatment with (Xba1/Not1), thereby obtaining pKOV-dhaT::trc-puuC, which was then introduced into the puuC::trc-puuC ydcW::trc-ydcW strain produced in Example 1-3 using the electroporation method described above, thereby producing the GSC123 puuC::trc-puuC ydcW::trc-ydcW dhaT::trc-puuC strain.
  • Example 5 Construction of puuC::trc-puuC ydcW::trc-ydcW dhaT::trc-puuC yqhD::trc-ydcW strain
  • trc-ydcW was inserted into the yqhD site of the byproduct-producing gene present in the genome.
  • the trc-ydcW was amplified as SEQ ID NO: 29/30
  • the left homologous arm was amplified with the SEQ ID NO: 31/32 primer
  • the right homologous arm was amplified with the SEQ ID NO: 33/34 primer
  • a homologous recombination site was produced through overlapping PCR using the SEQ ID NO: 29/34 primer.
  • the produced site was introduced into the pKOV plasmid and used for strain production.
  • the overlapping fragment was introduced into the pKOV plasmid after restriction enzyme treatment with (Xba1/Not1), thereby obtaining pKOV-yqhD::trc-ydcW, which was then introduced into the strain produced in Example 4 using the electroporation method described above, thereby producing the GSC123 puuC::trc-puuC ydcW::trc-ydcW dhaT::trc-puuC yqhD::trc-ydcW strain.
  • Example 6 Construction of puuC::trc-puuC ydcW::trc-ydcW dhaT::trc-puuC yqhD::trc-ydcW glpK::trc-puuC strains
  • trc-puuC was inserted into the byproduct-producing gene glpK site in the genome, and trc-puuC was amplified as SEQ ID NO: 35/36.
  • the homologous left arm was amplified with the SEQ ID NO: 37/38 primers, and the homologous right arm was amplified with the SEQ ID NO: 39/40 primers.
  • a homologous recombination site was produced through overlapping PCR using the SEQ ID NO: 35/40 primers. The produced site was introduced into the pKOV plasmid and used for strain production.
  • the overlapping fragment was introduced into the pKOV plasmid after restriction enzyme treatment with (BamH1/Not1), thereby obtaining pKOV-glpK::trc-puuC, and then introduced into the strain produced in Example 5 using the electroporation method described above, thereby producing the GSC123 puuC::trc-puuC ydcW::trc-ydcW dhaT::trc-puuC yqhD::trc-ydcW glpK::trc-puuC strain.
  • Example 7 Construction of puuC::trc-puuC ydcW::trc-ydcW dhaT::trc-puuC yqhD::trc-ydcW glpK::trc-puuC ldhA::trc-ydcW strains
  • trc-ydcW was inserted into the ldhA site of the byproduct-producing gene present in the genome, and trc-ydcW was amplified as SEQ ID NO: 41/42, the left homologous arm was amplified with the SEQ ID NO: 43/44 primer, and the right homologous arm was amplified with the SEQ ID NO: 45/46 primer.
  • a homologous recombination site was produced through overlapping PCR using the SEQ ID NO: 41/46 primer. The produced site was introduced into the pKOV plasmid and used for strain production.
  • the overlapping fragment was introduced into the pKOV plasmid after restriction enzyme treatment with (BamH1/Not1), thereby obtaining pKOV-ldhA::trc-ydcW, which was then introduced into the strain produced in Example 6 using the electroporation method described above, thereby producing the GSC123 puuC::trc-puuC ydcW::trc-ydcW dhaT::trc-puuC yqhD::trc-ydcW glpK::trc-puuC ldhA::trc-ydcW strain.
  • Example 8 Construction of puuC::trc-puuC ydcW::trc-ydcW dhaT::trc-puuC yqhD::trc-ydcW glpK::trc-puuC ldhA::trc-ydcW acs::trc-acs strains
  • trc-acs was inserted into the acs position, which is an acetyl CoA synthetase gene present in the genome, and the left homologous region of trc-acs was obtained using the primer of sequence number (48/49), and the right homologous region was obtained using the primer of sequence number (50/51) (see sequence number 47 for the base sequence of the acs gene).
  • the GSC123/pTrc99C strain was constructed by inserting the pTrc99C empty vector into the GSC123 strain.
  • the empty vector has no function and exhibits the same performance as the wild-type strain.
  • the recombinant strain in which the puuC gene and the ydcW gene were arranged in that order as shown in Fig. 2 and the trc promoter was used in front of each gene, had the best 3-HP production ability in a 24-hour culture.
  • the strain of Comparative Example 1 and the wild-type strain did not produce 3-hydroxypropionic acid at all, and it was confirmed that production of 3-hydroxypropionic acid was possible only when the puuC gene or ydcW gene was overexpressed, as in the recombinant strains of Examples 1 to 3. However, it was also found that 3-hydroxypropionic acid was not produced when both promoters were native promoters even if the puuC gene or ydcW gene was present in the genome.
  • nat represents the native promoter and trc represents the trc promoter.
  • Sequence number 1 is the base sequence of the puuC gene.
  • Sequence number 2 is the base sequence of the ydcW gene.
  • Sequence number 3 is the base sequence of a primer for amplifying the left arm of the trc-puuC gene of Example 1 by PCR. In addition, Sequence number 3 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 4 is the base sequence of a primer for amplifying the left arm of the trc-puuC gene of Example 1 by PCR.
  • Sequence number 5 is the base sequence of a primer for amplifying the right arm of the trc-puuC gene of Example 1 by PCR.
  • Sequence number 6 is the base sequence of a primer for amplifying the right arm of the trc-puuC gene of Example 1 by PCR.
  • sequence number 6 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 7 is the base sequence of a primer for amplifying the left arm of the trc-ydcW gene of Example 1 by PCR.
  • sequence number 7 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 8 is the base sequence of a primer for amplifying the left arm of the trc-ydcW gene of Example 1 by PCR.
  • Sequence number 9 is the base sequence of a primer for amplifying the right arm of the trc-ydcW gene of Example 1 by PCR.
  • Sequence number 10 is the base sequence of a primer for amplifying the right arm of the trc-ydcW gene of Example 1 by PCR.
  • sequence number 10 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 11 is the base sequence of a primer for amplifying the trc-puuC gene of Example 2 by PCR. In addition, Sequence number 11 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 12 is the base sequence of a primer for amplifying the trc-puuC gene of Example 2 by PCR.
  • Sequence number 13 is the base sequence of a primer for amplifying the left arm of the trc-puuC gene of Example 2 by PCR.
  • Sequence number 14 is the base sequence of a primer for amplifying the left arm of the trc-puuC gene of Example 2 by PCR.
  • Sequence number 15 is the base sequence of a primer for amplifying the right arm of the trc-puuC gene of Example 2 by PCR.
  • Sequence number 16 is the base sequence of a primer for amplifying the right arm of the trc-puuC gene of Example 2 by PCR. In addition, Sequence number 16 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 17 is the base sequence of a primer for amplifying the trc-ydcW gene of Example 3 by PCR.
  • sequence number 17 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 18 is the base sequence of a primer for amplifying the trc-ydcW gene of Example 3 by PCR.
  • Sequence number 19 is the base sequence of a primer for amplifying the left arm of the trc-ydcW gene of Example 3 by PCR.
  • Sequence number 20 is the base sequence of a primer for amplifying the left arm of the trc-ydcW gene of Example 3 by PCR.
  • Sequence number 21 is the base sequence of a primer for amplifying the right arm of the trc-ydcW gene of Example 3 by PCR.
  • Sequence number 22 is the base sequence of a primer for amplifying the right arm of the trc-ydcW gene of Example 3 by PCR. In addition, Sequence number 22 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 23 is the base sequence of a primer for amplifying the trc-puuC gene of Example 4 by PCR. In addition, Sequence number 23 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 24 is the base sequence of a primer for amplifying the trc-puuC gene of Example 4 by PCR.
  • Sequence number 25 is the base sequence of a primer for amplifying the left arm of the trc-puuC gene of Example 4 by PCR.
  • Sequence number 26 is the base sequence of a primer for amplifying the left arm of the trc-puuC gene of Example 4 by PCR.
  • Sequence number 27 is the base sequence of a primer for amplifying the right arm of the trc-puuC gene of Example 4 by PCR.
  • Sequence number 28 is the base sequence of a primer for amplifying the right arm of the trc-puuC gene of Example 4 by PCR. In addition, Sequence number 28 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 29 is the base sequence of a primer for amplifying the trc-ydcW gene of Example 5 by PCR.
  • sequence number 29 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 30 is the base sequence of a primer for amplifying the trc-ydcW gene of Example 5 by PCR.
  • Sequence number 31 is the base sequence of a primer for amplifying the left arm of the trc-ydcW gene of Example 5 by PCR.
  • Sequence number 32 is the base sequence of a primer for amplifying the left arm of the trc-ydcW gene of Example 5 by PCR.
  • Sequence number 33 is the base sequence of a primer for amplifying the right arm of the trc-ydcW gene of Example 5 by PCR.
  • Sequence number 34 is the base sequence of a primer for amplifying the right arm of the trc-ydcW gene of Example 5 by PCR. In addition, Sequence number 34 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 35 is the base sequence of a primer for amplifying the trc-puuC gene of Example 6 by PCR. In addition, Sequence number 35 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 36 is the base sequence of a primer for amplifying the trc-puuC gene of Example 6 by PCR.
  • Sequence number 37 is the base sequence of a primer for amplifying the left arm of the trc-puuC gene of Example 6 by PCR.
  • Sequence number 38 is the base sequence of a primer for amplifying the left arm of the trc-puuC gene of Example 6 by PCR.
  • Sequence number 39 is the base sequence of a primer for amplifying the right arm of the trc-puuC gene of Example 6 by PCR.
  • Sequence number 40 is the base sequence of a primer for amplifying the right arm of the trc-puuC gene of Example 6 by PCR. In addition, Sequence number 40 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 41 is the base sequence of a primer for amplifying the trc-ydcW gene of Example 7 by PCR.
  • sequence number 41 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 42 is the base sequence of a primer for amplifying the trc-ydcW gene of Example 7 by PCR.
  • Sequence number 43 is the base sequence of a primer for amplifying the left arm of the trc-ydcW gene of Example 7 by PCR.
  • Sequence number 44 is the base sequence of a primer for amplifying the left arm of the trc-ydcW gene of Example 7 by PCR.
  • Sequence number 45 is the base sequence of a primer for amplifying the right arm of the trc-ydcW gene of Example 7 by PCR.
  • Sequence number 46 is the base sequence of a primer for amplifying the right arm of the trc-ydcW gene of Example 7 by PCR.
  • sequence number 46 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 47 is the base sequence of the acs gene.
  • Sequence number 48 is the base sequence of a primer for amplifying the left arm of the trc-acs gene of Example 8 by PCR. In addition, Sequence number 48 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.
  • Sequence number 49 is the base sequence of a primer for amplifying the left arm of the trc-acs gene of Example 8 by PCR.
  • Sequence number 50 is the base sequence of a primer for amplifying the right arm of the trc-acs gene of Example 8 by PCR.
  • Sequence number 51 is the base sequence of a primer for amplifying the right arm of the trc-acs gene of Example 8 by PCR. In addition, Sequence number 51 is the base sequence of a primer for producing a fusion fragment by overlapping PCR of the amplified fragment.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La présente invention concerne un micro-organisme recombiné présentant une capacité accrue de production d'acide 3-hydroxypropionique, le micro-organisme présentant des voies de biosynthèse de l'acide 3-hydroxypropionique dans lesquelles une voie de conversion du 3-hydroxypropionaldéhyde en acide 3-hydroxypropionique et une voie de conversion de l'acétate en acétyl-CoA sont renforcées. En outre, la présente invention concerne un procédé de production d'acide 3-hydroxypropionique par l'utilisation du micro-organisme recombiné pour la production d'acide 3-hydroxypropionique.
PCT/KR2025/001440 2024-01-25 2025-01-24 Micro-organisme recombiné présentant une capacité accrue de production d'acide 3-hydroxypropionique Pending WO2025159558A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020240011673A KR20250116828A (ko) 2024-01-25 2024-01-25 3-하이드록시프로피온산 생산능이 증강된 재조합 미생물
KR10-2024-0011673 2024-01-25

Publications (1)

Publication Number Publication Date
WO2025159558A1 true WO2025159558A1 (fr) 2025-07-31

Family

ID=96545240

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2025/001440 Pending WO2025159558A1 (fr) 2024-01-25 2025-01-24 Micro-organisme recombiné présentant une capacité accrue de production d'acide 3-hydroxypropionique

Country Status (2)

Country Link
KR (1) KR20250116828A (fr)
WO (1) WO2025159558A1 (fr)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20210036632A (ko) * 2019-09-26 2021-04-05 주식회사 엘지화학 3-하이드록시프로피온산의 생산량이 증가된 미생물 및 이를 이용한 3-히드록시프로피온산의 생산 방법
KR20230081847A (ko) * 2021-11-30 2023-06-08 주식회사 엘지화학 포도당으로부터 3-하이드록시프로피온산을 생산하는 방법

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20210036632A (ko) * 2019-09-26 2021-04-05 주식회사 엘지화학 3-하이드록시프로피온산의 생산량이 증가된 미생물 및 이를 이용한 3-히드록시프로피온산의 생산 방법
KR20230081847A (ko) * 2021-11-30 2023-06-08 주식회사 엘지화학 포도당으로부터 3-하이드록시프로피온산을 생산하는 방법

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
KANCHANA R. KILDEGAARD, NIELS B. JENSEN, KONSTANTIN SCHNEIDER, EIK CZARNOTTA, EMRE ÖZDEMIR, TOBIAS KLEIN, JéRôME MAURY, BIRGITT: "Engineering and systems-level analysis of Saccharomyces cerevisiae for production of 3-hydroxypropionic acid via malonyl-CoA reductase-dependent pathway", MICROBIAL CELL FACTORIES, vol. 15, no. 1, 1 December 2016 (2016-12-01), XP055434334, DOI: 10.1186/s12934-016-0451-5 *
WANG XIAODI, CUI ZHENZHEN, SUN XI, WANG ZHIWEN, CHEN TAO: "Production of 3-Hydroxypropionic Acid from Renewable Substrates by Metabolically Engineered Microorganisms: A Review", MOLECULES, MDPI AG, CH, vol. 28, no. 4, CH , pages 1888, XP093337337, ISSN: 1420-3049, DOI: 10.3390/molecules28041888 *
ZHAO PENG; MA CHUNLU; XU LIDA; TIAN PINGFANG: "Exploiting tandem repetitive promoters for high-level production of 3-hydroxypropionic acid", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, SPRINGER BERLIN HEIDELBERG, BERLIN/HEIDELBERG, vol. 103, no. 10, 29 March 2019 (2019-03-29), Berlin/Heidelberg, pages 4017 - 4031, XP036768605, ISSN: 0175-7598, DOI: 10.1007/s00253-019-09772-5 *

Also Published As

Publication number Publication date
KR20250116828A (ko) 2025-08-04

Similar Documents

Publication Publication Date Title
WO2019203435A1 (fr) Nouveau promoteur issu d'une levure résistant aux acides organiques et procédé d'expression d'un gène cible à l'aide de celui-ci
WO2019117673A2 (fr) Nouveau polypeptide et procédé de production d'imp l'utilisant
WO2019117671A1 (fr) Micro-organisme producteur de 5'-inosine monophosphate et procédé de production de 5'-inosine monophosphate l'utilisant
WO2021167414A1 (fr) Micro-organisme producteur de nucléotide de purine et procédé de production de nucléotide de purine l'utilisant
WO2022050671A1 (fr) Micro-organismes produisant de la l-valine et procédé de production de l-valine les utilisant
WO2012046924A1 (fr) Souche productrice de xylitol dans laquelle une voie métabolique de l'arabinose est introduite, et procédé de production du xylitol l'employant
WO2022239953A1 (fr) Micro-organisme ayant une activité améliorée de 3-méthyl-2-oxobutanoate hydroxyméthyltransférase et ses utilisations
WO2015093831A1 (fr) Micro-organisme recombiné ayant une productivité accrue de d(-)2,3-butanediol, et procédé de production de d(-)2,3-butanediol l'utilisant
WO2015163682A1 (fr) Micro-organisme recombinant possédant une capacité améliorée à produire du 2,3-butanediol et procédé de production de 2,3-butanediol au moyen de celui-ci
WO2019235680A1 (fr) Micro-organisme produisant de l'acide 5'-xanthylique et procédé de production d'acide 5'-xanthylique au moyen de celui-ci
WO2013103246A2 (fr) Microorganisme recombinant produisant de l'acide quinolinique et procédé de production d'acide quinolinique l'utilisant
WO2020075943A1 (fr) Micro-organisme mutant produisant de l'acide succinique dans lequel une malate déshydrogénase à forte activité est introduite et procédé de préparation d'acide succinique à l'aide de celui-ci
WO2022191633A1 (fr) Nouveau variant de citrate synthase et procédé de production d'o-acétyl-l-homosérine ou de l-méthionine l'utilisant
WO2022098163A1 (fr) Micro-organismes ayant la capacité de produire de l'acide 3-hydroxypropionique à partir de glucose et leurs utilisations
WO2022245176A1 (fr) Micro-organisme produisant un nucléotide de purine, et procédé de production de nucléotide de purine au moyen de celui-ci
WO2015060649A1 (fr) Microorganisme produisant une o-succinyl homosérine et procédé de production d'o-succinyl homosérine à l'aide dudit microorganisme
WO2022055094A1 (fr) Micro-organisme recombinant pour la production d'acide l-glutamique et procédé de production d'acide l-glutamique l'utilisant
WO2018208120A2 (fr) Nouvelle souche de micro-organisme pour le métabolisme à haute performance de source de carbone dérivée de biomasse
WO2025159558A1 (fr) Micro-organisme recombiné présentant une capacité accrue de production d'acide 3-hydroxypropionique
WO2021235775A1 (fr) Nouveau polypeptide et procédé de production de l-leucine l'utilisant
WO2023277307A1 (fr) Souche pour la production d'acide l-glutamique hautement concentré et procédé de production d'acide l-glutamique l'utilisant
WO2024144283A1 (fr) Micro-organisme dans lequel de la glutamine synthétase hétérologue est introduite, et procédé de production de l-tryptophane à l'aide de celui-ci
WO2022191630A1 (fr) Nouveau variant de citrate synthase et procédé de production de l-valine l'utilisant
WO2022191635A1 (fr) Nouveau variant de citrate synthase et procédé de production d'acides l-aminés l'utilisant
WO2024167326A1 (fr) Micro-organisme recombiné pour la production d'acide 3-hydroxypropionique

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 25745380

Country of ref document: EP

Kind code of ref document: A1