WO1999028481A1 - Production microbienne d'hydroxyacetone et de 1,2-propanediol - Google Patents

Production microbienne d'hydroxyacetone et de 1,2-propanediol Download PDF

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Publication number
WO1999028481A1
WO1999028481A1 PCT/US1998/025318 US9825318W WO9928481A1 WO 1999028481 A1 WO1999028481 A1 WO 1999028481A1 US 9825318 W US9825318 W US 9825318W WO 9928481 A1 WO9928481 A1 WO 9928481A1
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Prior art keywords
genetically
yeast
acetol
engineered yeast
propanediol
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PCT/US1998/025318
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English (en)
Inventor
Douglas C. Cameron
Anita J. Shaw
Michael L. Hoffman
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Wisconsin Alumni Research Foundation
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Publication date
Priority claimed from US08/801,344 external-priority patent/US6087140A/en
Application filed by Wisconsin Alumni Research Foundation filed Critical Wisconsin Alumni Research Foundation
Priority to AU16107/99A priority Critical patent/AU1610799A/en
Publication of WO1999028481A1 publication Critical patent/WO1999028481A1/fr

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    • 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
    • C12P7/24—Preparation of oxygen-containing organic compounds containing a carbonyl group
    • C12P7/26—Ketones
    • 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
    • C12N9/88—Lyases (4.)
    • 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
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/18—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic polyhydric

Definitions

  • the invention is drawn to genetically-engineered yeast and their use in the production of acetol and 1,2-propanediol. More specifically, the present invention is drawn to genetically-engineered yeast having recombinant enzymatic activity which enables the yeast to ferment sugars into acetol and/or 1,2-propanediol in isolatable quantities.
  • 1,2-Propanediol (1,2-PD; also known as propylene glycol) is a major commodity chemical with an annual production greater than one billion pounds in the United States.
  • 1,2-PD is a major commodity chemical with an annual production greater than one billion pounds in the United States.
  • the major utilization of 1,2-PD is in unsaturated polyester resins, liquid laundry detergents, pharmaceuticals, cosmetics, antifreeze and de-icing formulations.
  • 1,2-PD is conventionally produced from petrochemicals. Unfortunately, several toxic chemicals, such as chlorine, propylene oxide, and propylene chlorohydrin are either required or are produced as by-products in the conventional synthesis. In the conventional route, 1,2-PD is produced by the hydration of propylene oxide, which is obtained from propylene. The synthetic process produces racemic 1,2-PD, an equimolar mixture of the two enantiomers. This chemical process has a number of disadvantages.
  • Propylene oxide is manufactured by one of two standard commercial processes: the chlorohydrin process or the hydroperoxide process.
  • the chlorohydrin process involves toxic chlorinated intermediates and the use of caustic or lime. Additionally, this process may result in air emissions of propylene chlorohydrin and chlorine. (Franklin
  • the hydroperoxide process involves oxidation of propylene by an organic hydroperoxide and results in the stoichiometric co-production of either tert- butanol or 1-phenyl ethanol. This make the economics of the production of propylene oxide via the hydroperoxide route directly related to the market for the co-produced byproducts. (Gait (1973).)
  • 1,2-PD is produced by several organisms when grown on exotic sugars. As early as 1937, the fermentation of L-rhamnose to 1,2-PD (later shown to be the S enantiomer) was described by Kluyver and Schnellen (1937). In E. coli and a variety of other microorganisms, L-rhamnose and L-fucose are metabolized to L- lactaldehyde and dihydroxyacetone phosphate. (Sawada and Takagi (1964) and
  • thermosaccharolyticum HG-8 (formerly Clostridium thermosaccharolyticum, ATCC 31960) also produces R-1,2-PD via methylglyoxal.
  • DHAP is converted to MG.
  • the MG is then reduced at the aldehyde group to yield acetol (i.e., hydroxyacetone).
  • the acetol is then further reduced at the ketone group to give R-1,2-PD.
  • acetol i.e., hydroxyacetone
  • R-1,2-PD the enzymes responsible for the production of 1,2-PD have not been isolated or cloned.
  • a first embodiment of the invention is a method of producing acetol and 1,2-PD by yeast fermentation.
  • the method comprises culturing a genetically-engineered yeast which expresses a recombinant enzyme which enables the yeast to produce acetol, 1,2-
  • 1,2-PD are produced by the yeast and secreted into the extracellular environment.
  • the preferred first embodiment of the invention is drawn to a method of producing acetol and 1,2-PD by fermentation using genetically-engineered yeast which comprises culturing a genetically-engineered yeast in a medium containing a suitable carbon source such as arabinose, glucose, galactose, lactose, xylose, sucrose, starch, and the like, wherein the yeast expresses a recombinant gene encoding methylglyoxal synthase.
  • a suitable carbon source such as arabinose, glucose, galactose, lactose, xylose, sucrose, starch, and the like
  • the yeast expresses a recombinant gene encoding methylglyoxal synthase.
  • the carbon source in the medium is fermented by the yeast into acetol and 1,2-PD, which can be isolated from the medium.
  • a second embodiment of the invention is drawn to genetically-engineered yeast which ferment a suitable carbon source into acetol and
  • the genetically-engineered yeast expresses one or more recombinant enzymes which enable the genetically-engineered yeast to produce acetol and/or 1,2-propanediol in isolatable quantities.
  • the genetically-engineered yeast express recombinant methylglyoxal synthase activity.
  • a third embodiment of the invention is drawn to a synthetic operon which enables the production of acetol and 1,2-PD in yeast.
  • the operon comprises one or more genes whose encoded gene products catalyze the formation of methylglyoxal in yeast and a promoter sequence functional in yeast operationally linked to the one or more genes.
  • the preferred synthetic operon comprises, in 5' to 3' order, a CUP1 promoter, operationally linked to a gene encoding methylglyoxal synthase, operationally linked to a CYC1 terminator.
  • the preferred embodiment of the invention is drawn to the use of genetically-engineered yeast, preferably genetically-engineered S. cerevisiae, which express recombinant methylglyoxal synthase activity to produce acetol and 1,2-PD.
  • the invention utilizes genetically-engineered yeast which express enzymes which enable the production of acetol and 1,2-PD from the fermentation of carbon sources utilizable by the yeast.
  • suitable or "utilizable” carbon sources refers to carbon sources utilizable by conventional and genetically-engineered yeast including, but not limited to, L-arabinose, D-glucose, D-galactose, D-xylose, lactose, sucrose, starch, and the like.
  • D-xylose as a utilizable carbon source
  • there is known genetically-engineered yeast which utilize xylose as a carbon source See, for instance, Ho et al. (1993).
  • a major advantage of the present invention is that microbial fermentation provides a clean and "environmentally friendly" synthetic route to acetol and 1,2-PD.
  • the microbial process can use as a substrate a renewable sugar such as glucose or xylose
  • Suitable carbon sources are also produced in commodity amounts from corn and sugar cane and from lignocellulosic biomass.
  • the microbial process produces no toxic wastes.
  • the byproducts of fermentation are carbon dioxide, alcohols, and organic acids, all of which can be purified as valuable co-products or used as animal feed.
  • Another distinct advantage of the invention is that it provides a unique route to acetol and 1,2-PD from readily-available sugars or starch. These carbon sources are cheap, renewable, and readily available.
  • a further advantage of the present invention is that microbial processes are straightforward to operate and do not involve high temperatures and pressures. Large fermentation facilities such as those used for the production of ethanol can be readily adapted to the production of acetol and 1,2-PD.
  • acetol and/or 1,2-PD from a sugar carbon source is favorable: on the order of about 1.0 moles or more of acetol or 1,2-PD per mole sugar.
  • 1,2-PD has very low toxicity to microorganisms. This allows for good cellular growth and viability at high final product titers. Cellular growth in the presence of 100 g/L 1,2-PD has been obtained.
  • Fig. 1 is a schematic representation of the production of 1,2-PD according to the present invention.
  • Fig. 2 is an HPLC elution profile of a standard solution of glucose, glycerol, 1,2- PD, 2,3-butandiol, and ethanol.
  • the HPLC protocol used was the same as that described in Examples 4-6.
  • Fig. 3 is an HPLC elution profile of a standard solution of glucose, succinate, acetate, acetol, and ethanol.
  • the HPLC protocol used was the same as that described in Examples 4-6.
  • Fig. 4 is an HPLC elution profile of culture medium taken from genetically- engineered yeast transformed and cultured as described in Example 4.
  • acetol and 1,2-PD display a retention times of 21.517 minutes and 20.417 minutes, respectively.
  • Fig. 5 is an HPLC elution profile of culture medium from the negative control yeast described in Example 3.
  • Fig. 6 is an ultraviolet spectrum of a standard acetol solution. The spectrum displays a characteristic absorption at 263.8 nm.
  • Fig. 7 is an ultraviolet spectrum of the isolated peak labelled "acetol" in Fig. 4. This ultraviolet spectrum confirms the identity of the 21.517 minute peak in the HPLC elution profile of Fig. 4 as being acetol.
  • Fig. 8 is a graph depicting the level of glucose (o), 1,2-PD (-), biomass (dry cell weight, DCW) (•) and acetol (x) over time in a culture of yeast transformed according to the present invention.
  • DHAP dihydroxyacetone phosphate
  • G-3-P glyceraldehyde-3-phosphate - O O
  • TPI triose phosphate isomerase
  • yeast explicitly encompasses, but is not limted to, microorganisms of the genus Saccharomyces, Schizosaccharomyces, Zygosaccharomyces, Pichia, Kluveromyces, Candida, Hansenula, Debaryomyces, and Torulopsis. Examples include Saccharomyces cerevisiae and Schizosaccharomyces pombe.
  • FIG. 1 An abbreviated schematic diagram of sugar metabolism resulting in the production of acetol and 1,2-PD according to the present invention is shown in Fig. 1. It is hypothesized that in yeast transformed according to the present invention, sugar is metabolized into DHAP and G-3-P by glycolytic enzymes common to most organisms. Together, DHAP and G-3-P are referred to as triose phosphates. Under normal conditions, the triose phosphates are interconverted by the activity of TPI.
  • DHAP is the initial intermediate in the acetol/ 1,2- PD pathway.
  • DHAP is converted to MG. While not being limited to a particular cellular mechanism, it is believed that in response to the increased intracellular level of MG, the transformed yeast detoxify the MG by a reductive pathway leading to acetol and then to 1 ,2-PD, which then diffuses or is actively secreted from the cell. Regardless of the actual mechanism, the present inventors have shown that by expressing recombinant MGS activity in yeast, the yeast will produce both acetol and 1,2-PD in isolatable quantities.
  • the crux of the invention is a method to produce acetol and/or 1,2-PD using genetically-engineered yeast which express recombinant enzyme activities whereby the yeast produce intracellular MG (or whereby the yeast produce increased amounts of intracellular MG as compared to non-transformed yeast).
  • the MG is then converted into acetol and 1,2-PD.
  • the acetol and 1,2-PD so formed may then be harvested from the cell culture medium.
  • the first step of the process is to identify and/or obtain the DNA sequences which encode the desired enzyme activities (i.e., activities leading to increased production of MG) and to insert them into the yeast. This can be accomplished by any means known to the art.
  • MGS methylglyoxal synthase
  • the gene which encodes the enzyme having the required activity is then incorporated into a suitable vector which is then used to transform a yeast host.
  • the preferred host is S. cerevisiae (baker's yeast).
  • the genetically- engineered yeast produce acetol and 1,2-PD from the fermentation of commonly utilized carbon sources, including arabinose, galactose, glucose, lactose, sucrose, xylose, and starch. Careful selection of mutant hosts can also be used to increase the yield of acetol and 1 ,2-PD.
  • a TPI knockout mutant or a mutant having altered TPI activity can be used as the host cell to increase the intracellular levels of MG, thereby increasing acetol and 1,2-PD production. See, for instance, Pompliano et al. (1990) for a discussion on altering the specificity of TPI.
  • glyoxalase knockout mutants can also be used as host cells, thereby increasing the intracellular level of MG for conversion to acetol and
  • Appropriate host selection also allows the conditions under which 1,2-PD is produced to be varied and/or optimized, e.g., aerobic or anaerobic production, different substrates as a carbon source, etc.
  • Isolation of the acetol and 1,2-PD formed from the cell medium can be accomplished by any means known in the separation art.
  • the preferred method is to filter the culture medium to separate cells and cellular debris, and then to isolate the acetol and 1,2-PD from the medium by vacuum distillation. (See, for instance, Simon et al. (1987).) If so desired, the yeast may be completely lysed by any known means prior to isolation of the products.
  • yeast transformed to contain the preferred vector which contains an insert encoding green fluorescence protein rather than MGS do not produce acetol or 1,2-PD. See Example 3 and Fig. 5.
  • YpJ66 is constructed from YEp352, whose oligonucleotide sequence is shown in SEQ.
  • the vector is then transformed into YPH500 (ATCC 76626) (leu , trp ' , urd, lys ' , ode ' , his ' ) by standard methods and fed the required amino acids for growth, except uracil, which is used as the marker to maintain the plasmid in yeast.
  • Yeast transformed to contain the mgs insert produce acetol and 1,2-PD in isolatable quantities when fermented on a wide variety of substrates. Any vector capable of successfully transforming yeast to express the required enzyme activity can be used in the present invention.
  • the vector may be an integrating plasmid, in which case the recombinant gene is incorporated into the genome of the yeast host, or the vector may be replicating, in which case the recombinant gene might be present only on one or more copies of a self-replicating plasmid, or the required genetic elements may be placed on a yeast artificial chromosome (YAC).
  • YAC yeast artificial chromosome
  • yeast auxotrophic markers such as HIS3 (imidazole glycerolphosphate dehydratase), LEU2 ( ⁇ -isopropylmalate dehydrogenase), LYS2 ( ⁇ - aminoadipate reductase), TRP1 (N-(5'-phosphoribosyl)-anthranilate isomerase), and URA3 (orotidine-5'-phosphate decarboxylase). Selection is accomplished by culturing the yeast in a suitable media lacking the required nutrient.
  • HIS3 imidazole glycerolphosphate dehydratase
  • LEU2 ⁇ -isopropylmalate dehydrogenase
  • LYS2 ⁇ - aminoadipate reductase
  • TRP1 N-(5'-phosphoribosyl)-anthranilate isomerase
  • URA3 orotidine-5'-phosphate decarboxylase
  • Replicating vectors include an autonomously replicating sequence (ARS) or a 2 ⁇ sequence to allow multiple copies of the plasmid to be replicated within each yeast cell.
  • ARS autonomously replicating sequence
  • 2 ⁇ sequence to allow multiple copies of the plasmid to be replicated within each yeast cell.
  • the vector may also include a centromere sequence (CEN), which will generally limit the copy number of the plasmid vector to one or two per cell.
  • CEN centromere sequence
  • YAC's which further include telomere sequences, may also be used to carry the recombinant genes necessary to induce the production of acetol and 1,2-PD by the transformed yeast.
  • the DNA coding region for E. coli. MGS shown in SEQ. ID. NO: 1, was amplififed by the polymerase chain reaction (PCR) using conventional and well known techniques.
  • the PCR primers used are listed in SEQ. ID. NO: 2 (a Kpnl restriction site is defined by nucleotides 3-8: GGTACC) and SEQ. ID. NO: 3 (a EcoRI restriction site is defined by nucleotides 3-8: GAATTC).
  • the nucleotide base sequence of the amplified fragment was confirmed by sequencing using conventional methods.
  • YpJ66 is based on YEp352 and contains the CUP1 promoter and the CYC1 terminator for the expression of proteins in S. cerevisiae.
  • YpJ66 is constructed from YEp352, whose oligonucleotide sequence is shown in SEQ. ID. NO: 4, according to the method of Hill et al. (1986). This is accomplished by inserting the CUP1 promoter, (galK), and CYC1 terminator sequence into the Xbal site of Yep352.
  • YEP352 also contains the URA3 gene wich allows selection in a URA3- host strain.
  • YEp352 SEQ. ID. NO: 4
  • the EcoRI, Kpnl, Smal, and Hindlll sites are knocked out using T4 DNA polymerase.
  • a BamHI/EcoRI fragment of the genomic CUP1 promoter is ligated at the EcoRI site to a Clal/EcoRI fragment of the CYC1 terminator.
  • This fragement is blunt-ended (BamHI and Clal blunt-ended with T4 polymerase) into the blunt-ended (T4 polyerase) Xbal site in YEp352, with the filled-in BamHI site of the insert furthest from the BamHI site in
  • E. coli DNA was subjected to PCR using the primers depicted in SEQ. ID NOS: 2 and 3.
  • the PCR amplification was digested with EcoRI/KpnI.
  • the fragments were separated on an agarose gel.
  • YpJ66 was also digested with EcoRI/KpnI and the fragments separated on an agarose gel.
  • the digested PCR fragment was then ligated to the large fragment from YpJ66. This plasmid was named pMH36.
  • the CUP1 promoter is induced by copper and other heavy metals in fermentation media (Etcheverry, 1990).
  • the pMH36 construct was designed so that the coding region of MGS is downstream from the CUPl promoter (i.e., in the 3' direction from CUPl) and upstream from the CYCl terminator (i.e., in the 5' direction from CYCl).
  • the identity of the pMH36 construct was confirmed by restriction enzyme analysis on agarose gel.
  • the complete nucleotide sequence of pMH36 is provided in SEQ. ID. NO: 5. All plasmid construction steps were performed in E. coli strain AG1 (Stratagene,
  • Example 2 - Yeast Transformation Yeast strain YPH500 (ATCC 76626) (MAT ⁇ ura3 lys2 ade2 trpl his3 leu2) was used for all of the Examples (Sikorski & Hieter, 1989). Yeast were transformed according to the lithium acetate method of Ito as modified by Kaiser et al. , 1994:
  • Yeast cultures were grown overnight in the medium defined below to an OD 600 of from about 0.3 to about 0.5. The cultures were then centrifuged at 4000 x g for 5 minutes and the cells resuspended in 10 mL sterile water. The cells were then centrifuged at 5000 x g for 5 minutes and the cells resuspended in 1.5 mL of a buffered lithium acetate solution (1 vol. lOx Tris/EDTA buffer, pH 7.5; 1 vol. lOx LiAc (1 M), pH 7.5; 8 vols. sterile water) and incubated for 1 hour at 30°C.
  • a buffered lithium acetate solution (1 vol. lOx Tris/EDTA buffer, pH 7.5; 1 vol. lOx LiAc (1 M), pH 7.5; 8 vols. sterile water
  • yeast strain YPH500 was transformed with a plasmid designated pMHl.
  • pMHl encodes (and expresses) green fluoresence protein in the same cassette as pMH36 (CUPl promoter and CYCl terminator).
  • Fig. 5 shows an HPLC elution profile of culture medium from a culture of yeast transformed with pMHl. No acetol or 1,2-PD is evident in the elution profile. Yeast were cultured and HPLC analyses were performed as detailed in Examples 4-6.
  • SDM synthetic defined medium
  • MnSO 4 0.4 mg/L; Na 2 MoO 4 , 0.2 mg/L; ZnSO 4 , 0.4 mg/L; biotin, 2 mg/L; calcium pantothenate, 0.4 mg/L; folic acid, 2 mg/L; nicotinic acid, 0.4 mg/L; /.-aminobenzoic acid, 0.2 mg/L; pyridoxine hydrochloride, 0.4 mg/L; riboflavin, 0.2 mg/L; and thiamine hydrochloride, 0.4 mg/L.
  • This media was supplemented with the required nutrients (lysine, adenine, tryptophan, histidine and leucine) to support the auxotrophs and maintain the plasmid (Kaiser et al., 1994). Copper (as CuCl 2 ) was added to the medium to induce the expression of MGS.
  • Anaerobic fermentations were performed in 15 mL anaerobic tubes; aerobic fermentations were performed in 250 mL baffled shake flasks. All fermentations were agitated by mixing at 200 rpm and maintained at 30°C.
  • Figs. 2 and 3 HPLC elution profiles for standard solutions using the above protocol are depicted in Figs. 2 and 3.
  • Fig. 2 is an HPLC elution profile of a standard solution containing glucose, glycerol, 1,2-PD, 2,3-butandiol, and ethanol.
  • Fig. 3 is an HPLC elution profile of a standard solution containing glucose, succinate, acetate, acetol, and ethanol.
  • FIG. 4 An HPLC elution profile of the culture medium from this Example is presented in Fig. 4. As shown in Fig. 4, 1,2-PD eluted from the column at 20.417 minutes, which is the same retention time displayed by the 1,2-PD standard shown in Fig. 2. In Fig. 4, acetol eluted at 21.517 minutes, which is slightly faster than the 21.650 retention time displayed by the acetol standard shown in Fig. 3.
  • Fig. 4 To confirm the identity of the 21.517 minute peak in Fig. 4 as being acetol, the peak was collected and analyzed by ultraviolet spectroscopy against an acetol standard.
  • the U.V. spectrum for the acetol standard is depicted in Fig. 6.
  • Fig. 6 displays a very characteristic U.V. absorption at 263.8 nm.
  • the U.V. spectrum for the 21.517 minute peak of Fig. 4 is shown in Fig. 7. These two U.V. spectra are virtually identical, indicating that the 21.517 minute peak is, in fact, acetol.
  • Fig. 8 glucose ( ⁇ ), 1,2-PD (-), biomass (•) and acetol (x). Each data point is the average of three experiments. Over the course of time, the concentration of acetol (x) and 1,2-PD ( ⁇ ) increases markedly. Concurrently, the concentration of glucose drops quickly. This graph shows that as the yeast consume the glucose, they convert this carbon source to acetol and 1,2- PD, which are then secreted into the culture medium.
  • the invention is not limited to the particular protocols, reagents, host strains, plasmids, and promoters described herein, but encompasses all modified forms thereof which are encompassed by the attached claims.

Abstract

La présente invention concerne une levure élaborée génétiquement transformant par fermentation les sources de carbone utilisables en acétol et/ou en 1,2-propanédiol, des procédés de production de l'acétol et du 1,2-propanédiol par fermentation en utilisant la levure transformée, ainsi que des opérons synthétiques permettant d'effectuer cette transformation.
PCT/US1998/025318 1997-02-19 1998-11-30 Production microbienne d'hydroxyacetone et de 1,2-propanediol WO1999028481A1 (fr)

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AU16107/99A AU1610799A (en) 1997-12-03 1998-11-30 Microbial production of hydroxyacetone and 1,2-propanediol

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US08/801,344 US6087140A (en) 1997-02-19 1997-02-19 Microbial production of 1,2-propanediol from sugar
US98471797A 1997-12-03 1997-12-03
US08/984,717 1997-12-03

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008116851A1 (fr) * 2007-03-23 2008-10-02 Metabolic Explorer Micro-organisme obtenu par génie métabolique utile pour produire acétol
WO2011012697A2 (fr) 2009-07-30 2011-02-03 Metabolic Explorer Enzyme yqhd mutante pour la production d'un produit biochimique par fermentation
WO2011012702A1 (fr) 2009-07-30 2011-02-03 Metabolic Explorer Glycérol déshydrogénase (glydh) mutante pour la production d'un agent biochimique par fermentation
WO2011012693A1 (fr) 2009-07-30 2011-02-03 Metabolic Explorer Méthylglyoxal synthétase (mgs) mutante pour la production d'un agent biochimique par fermentation
WO2012172050A1 (fr) 2011-06-15 2012-12-20 B.R.A.I.N. Biotechnology Research And Information Network Ag Nouveaux moyens et procédés de production de propanediol
US9617567B2 (en) 2008-11-07 2017-04-11 Metabolic Explorer Use of sucrose as substrate for fermentative production of 1,2-propanediol

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WO1998037204A1 (fr) * 1997-02-19 1998-08-27 Wisconsin Alumni Research Foundation Production microbienne de 1,2-propanediols a partir de sucre

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008116851A1 (fr) * 2007-03-23 2008-10-02 Metabolic Explorer Micro-organisme obtenu par génie métabolique utile pour produire acétol
US9617567B2 (en) 2008-11-07 2017-04-11 Metabolic Explorer Use of sucrose as substrate for fermentative production of 1,2-propanediol
WO2011012697A2 (fr) 2009-07-30 2011-02-03 Metabolic Explorer Enzyme yqhd mutante pour la production d'un produit biochimique par fermentation
WO2011012702A1 (fr) 2009-07-30 2011-02-03 Metabolic Explorer Glycérol déshydrogénase (glydh) mutante pour la production d'un agent biochimique par fermentation
WO2011012693A1 (fr) 2009-07-30 2011-02-03 Metabolic Explorer Méthylglyoxal synthétase (mgs) mutante pour la production d'un agent biochimique par fermentation
WO2012172050A1 (fr) 2011-06-15 2012-12-20 B.R.A.I.N. Biotechnology Research And Information Network Ag Nouveaux moyens et procédés de production de propanediol

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