WO2008144792A1 - Procédé pour produire de l'acide ascorbique comprenant l'utilisation de pectobacterium cypripedit - Google Patents

Procédé pour produire de l'acide ascorbique comprenant l'utilisation de pectobacterium cypripedit Download PDF

Info

Publication number
WO2008144792A1
WO2008144792A1 PCT/AT2008/000185 AT2008000185W WO2008144792A1 WO 2008144792 A1 WO2008144792 A1 WO 2008144792A1 AT 2008000185 W AT2008000185 W AT 2008000185W WO 2008144792 A1 WO2008144792 A1 WO 2008144792A1
Authority
WO
WIPO (PCT)
Prior art keywords
gluconic acid
diketo
glucose
acid
keto
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.)
Ceased
Application number
PCT/AT2008/000185
Other languages
German (de)
English (en)
Other versions
WO2008144792A4 (fr
Inventor
Claudia Pacher
Klaus D. Kulbe
Eva Steiner
Günter REMBART
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.)
Vogelbusch GmbH
Original Assignee
Vogelbusch GmbH
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 Vogelbusch GmbH filed Critical Vogelbusch GmbH
Publication of WO2008144792A1 publication Critical patent/WO2008144792A1/fr
Publication of WO2008144792A4 publication Critical patent/WO2008144792A4/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
    • C12P17/02—Oxygen as only ring hetero atoms
    • C12P17/04—Oxygen as only ring hetero atoms containing a five-membered hetero ring, e.g. griseofulvin, vitamin C
    • 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
    • C12P7/58—Aldonic, ketoaldonic or saccharic acids
    • C12P7/60—2-Ketogulonic acid

Definitions

  • the present invention relates to a process for producing ascorbic acid, in which in a first step, a carbon source is fermentatively converted to 2,5-diketo-D-gluconic acid, which then in a second step enzymatically with 2,5-diketo-D-gluconic acid Corynebacterium glutamicum reductase is converted to 2-keto-L-gulonic acid.
  • Ascorbic acid is used in the pharmaceutical and food industries as a vitamin and antioxidant.
  • L-ascorbic acid is currently produced predominantly according to the process developed by Reichstein (1934). After this manufacturing process of Reichstein obtained vitamin C from D-glucose in a chemical-biocatalytic process. The glucose is first reduced to sorbitol and then oxidized with sorbose bacteria to the carbohydrate L-sorbose. The sorbose is further oxidized with the addition of acetone, wherein after the subsequent cleavage of the acetone and dehydration, the vitamin C is formed.
  • the Reichstein synthesis requires not only high energy and chemical expenditure, but also sophisticated wastewater treatment in order to avoid the release of lindane-like by-products. Furthermore, this process provides only a yield of about 50%.
  • Sonoyama et al. U.S. Patent 4,879,229, Shionogi & Co., Ltd., Japan, 1989 disclose the use of facultative anaerobic strains of the genus Pectobacter to produce 2,5-DKG.
  • a 2,5-DKG-producing microorganism of the genus Pectobacter or any ingredient thereof (such as enzyme extracts) or immobilized cells is brought into contact with D-glucose.
  • EP 1141368B1 (Genencor International, Inc., California, 2006) describes the biocatalytic production of 2-KLG from a carbon source, wherein the co-factor required for the reaction is regenerated.
  • An alternative method is disclosed, for example, in U.S. Patent 4,757,012 (Genentech Inc. California) which discloses the purification and recombinant production of 2,5-diketo-gluconic acid (2,5-DKG) reductase and their use in the reductive conversion of 2,5-diketo-gluconic acid in 2-keto-L-gulonic acid (2-KLG).
  • the invention is therefore based on the object to provide a method of the type mentioned above, which on the one hand represents an economical alternative to the biotechnological methods described so far and on the other hand overcomes the disadvantages of the Reichstein process.
  • the invention is achieved in that the carbon source is glucose and the conversion of glucose into 2,5-diketo-D-gluconic acid by the Pectobacter cypripedii strain HEPOl (DSMZ 12393) directly.
  • the advantage of the present invention is that the invention Pectobacter cypripedii strain HEPOl can convert glucose directly into 2,5-diketo-D-gluconic acid (2.5 DKG) and thus represents a further development of the prior art. So far, it was by means of Pectobacter spp. it is only possible to ferment gluconic acid, which is considerably more expensive than glucose (gluconic acid costs about twice the amount of glucose), into the intermediate 2.5-DKG.
  • the new Pectobacter strain HEPO1 differs from the previous strain described in the prior art (DSMZ 30182) in that it is capable of nitrate reduction, of growth on malonic acid or of acid formation from L-rhamnose and D-maltose on the basis of physiological and biochemical characteristics is and also has a urease activity.
  • strain HEPO1 (DSMZ 12393) does not have starch hydrolysis activity.
  • the air supply in the first step, can be reduced or adjusted until anaerobic conditions in the nutrient solution are reached, if the formation of the 2,5-diketo-D-gluconic acid from the intermediate 2-keto-D-gluconic acid fails , This ensures that substantially all of the 2-keto-D-gluconic acid is converted to 2,5-diketo-D-gluconic acid, which is the starting substrate for subsequent reduction to 2-keto-L-gulonic acid, giving a higher total Product yield can be achieved.
  • the air supply can be controlled via an oxygen probe.
  • an oxygen probe By monitoring the oxygen content in the nutrient solution using an oxygen probe, it is achieved that, on the one hand, the oxygen content in the nutrient solution corresponds to the particular optimum of the microorganism used, and, on the other hand, that required for complete conversion of 2-keto-D-gluconic acid into 2,5-diketone. Controlled D-gluconic acid necessary anaerobic conditions.
  • the pH may be monitored and / or controlled during the fermentation of the first step.
  • the formation of gluconic acid leads to a lowering of the pH, whereby the growth of the microorganisms is suppressed.
  • By controlling the pH it is ensured that the pH of the nutrient solution is always within the optimal range for the growth of the nutrient solution Microorganisms and thus the healthy growth of microorganisms is maintained.
  • the first step of the method can be carried out in batch mode.
  • all ingredients are introduced into the fermenter and fermented in a conventional manner by the microorganisms.
  • the first step of the process can be carried out in fed-batch mode.
  • fed-batch operation a controlled substrate feed avoids an excessive lowering of the educt concentration, so that the microorganisms are always in the exponential growth phase.
  • the coenzyme NADPH required in the second step is regenerated by the glucose dehydrogenase (GDH), which converts glucose to gluconic acid, whereby the gluconic acid formed by the regeneration and optionally remaining glucose and / or or 2,5-diketo-D-gluconic acid of the process according to the invention is returned to the first step.
  • GDH glucose dehydrogenase
  • Continuous regeneration of the coenzyme NADPH may result in a lower coenzyme concentration than would be necessary for complete conversion of 2,5-DKG to 2-KLG without coenzyme regeneration.
  • Na citrate or Na acetate may be used as the buffer.
  • the Na citrate or Na acetate buffer not only the costs for the buffer system could be lowered, but also the enzyme activity could be increased compared to the Bis-Tris system described in the literature.
  • the gluconic acid and optionally glucose and / or 2,5-diketo-D-gluconic acid prior to recycling by chromatographic separation methods, such as ion exchange chromatography, from the 2- Keto-L-gulonic acid product stream are separated.
  • chromatographic separation methods such as ion exchange chromatography
  • the 2,5-diketo-D-gluconic acid reductase from Corynebacterium glutamicum can be produced recombinantly in E. coli using a pET vector.
  • CglakrPETF and CglakrPETR and the plasmid pPCl may be used as templates for the preparation of the insert DNA as primers for the PCR, the amplified fragment being inserted into the expression vector pET-21d.
  • coli BL21 (DE3) strain transformed with pET-21d can be incubated in a minimal medium MPC-Gly which contains in one liter 10 g peptone from casein, 10 g glycerol, 0.1 ml IM CaCl 2 , 1M MgSO 4 .7H 2 O and 200 ml MPC-Gly salts, with the MPC-Gly saline solution in one liter of water containing 15 g KH 2 PO 4 , 2.5 g NaCl and 5 g NH 4 Cl.
  • MPC-Gly which contains in one liter 10 g peptone from casein, 10 g glycerol, 0.1 ml IM CaCl 2 , 1M MgSO 4 .7H 2 O and 200 ml MPC-Gly salts, with the MPC-Gly saline solution in one liter of water containing 15 g KH 2 PO 4 , 2.5 g NaCl and 5 g NH 4 Cl.
  • reaction mixture is separated into a product stream and a biocatalyst stream by a membrane separation process. This ensures that the enzymes can be performed in the process cycle without major losses.
  • NADP + are retained by a charged ultrafiltration membrane.
  • the separation of the product stream and the biocatalyst stream through an ultrafiltration membrane causes the enzymes or cofactors to remain intact throughout the process
  • Enzyme reactor are present, and need not be constantly replaced by new enzymes or cofactors.
  • the production of ascorbic acid by the process of the present invention is carried out in a two-stage process.
  • glucose is fermentatively converted to 2,5-diketo-D-gluconic acid (2,5-DKG).
  • HEPOI Pectobacter cypripedii strain
  • TSA tryptone Soy Agar
  • the pH of the medium was adjusted to 7.02 using caustic soda (100 g / L). Then the medium was brought to a total volume of 250 ml with tap water.
  • 300 ml Erlenmeyer flasks each case 1.25 g of CaCO 3 were weighed and added to 50 ml of the above-described medium and sterilized for 20 minutes at 121 0 C. After cooling, the Erlenmeyer flasks were in each case inoculated by means of a loop with the Pectobacter cypripedii strain of the TSA plates. The Erlenmeyer flasks were then incubated for 16 hours on a shaking incubator at 200 rpm at 28 ° C. The resulting preculture is used for the subsequent fermentation.
  • the fermentation itself was carried out in a stirred tank fermenter with a total volume of 10 liters, which is equipped with a pH, ⁇ 2 , temperature and fermentation oil selector and associated measuring and control electronics. Furthermore, the CO 2 and O 2 content in the exhaust air of the fermenter was monitored.
  • Both batch and fed-batch fermentation were analyzed by conventional methods. These include the determination of the optical density, the enzymatic determination of glucose and gluconic acid and HPLC.
  • the optical density 1 ml of sample was admixed with 8 ml of citric acid monohydrate solution (40 g / l) and made up to 10 ml. The optical density was measured at 600 nm.
  • the glucose and gluconic acid were each treated with a commercially available enzyme assay determined photometrically at 340 nm (Boehringer, order numbers 716251 and 428191, respectively).
  • the fermentation can be carried out both as a batch fermentation and as a fed-batch fermentation.
  • a batch fermentation will be described.
  • mineral solution A 192.2 g of CSL (Com Steep Liquor), 1.35 g of MgSO 4 .7H 2 O and 8.96 g of K 2 HPO 4 were dissolved in 2503 g of tap water and sterilized at 121 ° C. for 20 minutes.
  • the preparation of the mineral solution B was carried out by slurry of 182.0 g of CaCO 3 in 1499.8 g of tap water and subsequent sterilization at 121 0 C for 20 minutes.
  • the sterile media (glucose solution, mineral solutions A and B) were introduced via a sterile funnel into the fermenter and the weight determined by reweighing.
  • the pH of the medium in the fermenter was adjusted to pH 7.0 by the addition of a NaOH solution (300 g / L).
  • the fermenter was inoculated with the contents of two pre-culture Erlenmeyer flasks obtained as described above.
  • the following parameters were set and kept as constant as possible during the entire fermentation period, with the exception of the amount of air, as described later:
  • the fermentation for the production of 2,5-DKG was carried out in another experiment than fed-batch process, which essentially corresponds to the batch process described above, but with the following changes.
  • the fermentation medium does not contain 200 g / l glucose but only 100 g / l glucose.
  • the sterilized stirred tank was filled with 5.5 liters (without inoculum) medium as described above and inoculated with Pectobacter cypripedii (2 preculture Erlenmeyer flasks).
  • the feed (feed, 50% glucose solution) was started when the CO 2 content in the exhaust air is above 1% by volume.
  • the feed rate of the feed solution is controlled by the CO 2 content in the exhaust air, with only so much glucose solution is supplied that the CO 2 content does not drop significantly below 1%.
  • the purified 2,5-DKG is enzymatically converted to 2-KLG by 2,5-diketo-D-gluconic acid reductase.
  • the 2,5-diketo-D-gluconic acid reductase from Corynebacterium glutamicum (DMSZ 20301) was isolated and amplified by PCR.
  • PCR reaction all relevant components were combined in a master mix for the amplification reaction whose composition is shown in Table 1 and then aliquoted into PCR tubes.
  • the primers used for the PCR have the following sequences:
  • Taq DNA polymerase (5 U / ⁇ l) 0.25
  • thermocycler T3, Biometra (Göttingen, Germany)
  • amplified under the following conditions (Table 2).
  • the amplified PCR product was purified using methods known in the art. After purification, the DNA insert was digested by the restriction enzymes BamHI and Xhol and cloned into the appropriately pretreated pET 21d vector (Novagen, Darmstadt, Germany) using the Topo TA Cloning Kit (Invitrogen).
  • the pET vectors are based on the T7 expression system, which is characterized by a very high transcription efficiency.
  • the system is derived from bacteriophage T7, which is produced by its strong promoters derived from the phage T7.
  • RNA polymerase successfully competes with the transcriptional machinery of the host.
  • the prepared vector (pET-21d) was transformed by heat shock transformation into E.
  • the E. coli strain BL21 (DE3) is particularly suitable for the expression of recombinant proteins in pET systems.
  • the T7 RNA polymerase is expressed after EPTG addition and subsequently the genes under the control of the T7 promoter are transcribed.
  • the pET system was therefore selected because it adds a C-terminal His tag to the desired DNA insert and thus makes very simple purification of the DNA insert possible by means of nickel chelate affinity chromatography.
  • the recombinant cells are then resuspended in a modified minimal medium (MPC-Gly medium) containing in one liter 10 g peptone from casein, 10 g glycerol, 0.1 ml IM CaCl 2 , 1 M MgSO 4 .7H 2 O, and 200 ml of MPC-Gly contains salts, the MPC-Gly saline solution in one liter of water containing 15 g KH 2 PO 4 , 2.5 g NaCl and 5 g NH 4 Cl. After the recombinant cells were cultured in said medium at 37 ° C.
  • MPC-Gly medium modified minimal medium containing in one liter 10 g peptone from casein, 10 g glycerol, 0.1 ml IM CaCl 2 , 1 M MgSO 4 .7H 2 O, and 200 ml of MPC-Gly contains salts, the MPC-Gly saline solution in one liter of water
  • the expression of the heterologous gene is induced by the addition of 5 g / l lactose and the cells are allowed to reach 25 0 C, pH 7.0 and pO 2 20% to grow until an optical density of 8.0 has been reached.
  • the recombinant cells were pelleted by centrifugation (15,000 g) for 20 minutes and then mechanically disrupted by homogenizer (Homogenizer Invensis APV-2000, 1000 bar) or French Pressure Cell (70 bar).
  • homogenizer Homogenizer Invensis APV-2000, 1000 bar
  • French Pressure Cell 70 bar
  • the medium is first charged with a transition metal ion (Ni 2+ ) to form a chelate before use.
  • the proteins bind to the medium, depending on the presence of surface histindin residues that have an affinity for the chelated metal ions. The bond strength is always determined by the metal ion and the pH of the buffer.
  • the bound protein can be eluted by competitive elution with imidazole.
  • the iminodiacetic acid ligand is coupled to the chelating sepharose and thus allows the exchange of immobilized metal ions.
  • the 2,5-diketo-D-gluconic acid reductase (DKR) thus prepared has an activity of about 200 U / l culture.
  • the DKR is used in soluble form.
  • 250 mM (58 g / l) 2,5-DKG, 2 U / ml DKR, 250 mM (45 g / l) glucose, 2 U / ml glucose dehydrogenase (GDH) are added to a 21 reaction vessel. and 0.26 mM (0.21 g / L) of NADP + added.
  • the volume in the batch reactor was 500 or 1000 mL.
  • This reaction gives under the conditions mentioned 56.0 g of 2-keto-L-gulonic acid / l reaction solution (ie buffer, either Na acetate or Na citrate or bis-Tris) (yield 96.7%) and 42.1 g Gluconic acid / 1 reaction solution (ie buffer, either Na acetate or Na citrate or Bis-Tris) (yield 93.5%).
  • the 2-keto-L-gulonic acid is subsequently separated from the gluconic acid, glucose and 2,5-DKG by an ion exchanger and used for further reaction in ascorbic acid.
  • the gluconic acid, glucose and 2,5-DKG are recycled in the first reaction step (fermentation of Pectobacter cypripedii).
  • the above-described two-stage process which proceeds in two spatially separate and non-interconnected reaction vessels, can be combined into a so-called hybrid process.
  • the stirred tank in which the fermentation of Pectobacter cypripedii HEPO1 and the concomitant conversion of glucose into 2,5-diketo-D-gluconic acid (steps 1 of the above method) expires
  • the enzyme reactor in which by the 2, 5-diketo-D-gluconic acid reductase catalyzed reaction takes place, directly connected via connecting lines.
  • An advantage of the hybrid method according to the invention is that continuous 2.5-DKG is formed in the fermenter which, after the microorganisms have been separated from the liquid stream of the fermenter by filtration, is passed into the enzyme reactor.
  • the enzymes necessary for continuous process control, DKR and GDH, and the coenzymes NADPH and NAPD + are retained in the enzyme reactor, while the unused substrates 2,5-diketo-D-glucose and glucose and the products gluconic acid and 2-keto-L- Gulonic acid can pass through the membrane unhindered.
  • the enzymes (DKR and GDH) are retained by the ultrafiltration membrane due to their size, whereas the coenzymes (NADPH and NADP + ) due to their negative charge on the passage are prevented.
  • the product of the process according to the invention must be 2-keto-L- Gulonic acid are separated from the product stream.
  • the ion exchange chromatography strongly basic AIEX was used to separate the 2-keto-L-gulonic acid from the glucose or gluconic acid.
  • the biocatalyst system (DKR, GDH and NADPH) remains through the ultrafiltration membrane in the enzyme reactor.
  • the vessels used for the hybrid method according to the invention are known in the prior art and are on the one hand a conventional stirred tank having the necessary measuring and control electronics such as pH electrode, pO 2 electrode, etc., and on the other hand, an enzyme reactor, the semipermeable membrane having.
  • the semipermeable membrane used is a charged ultrafiltration membrane (NTR-7430, Nitto Electric Industrial Co. (Japan)).
  • NTR-7430 Nitto Electric Industrial Co. (Japan)
  • the stirred tank the fermentative conversion of gluconic acid into 2,5-DKG by means of Pectobacter cypripedii HEPOl takes place, while in the enzyme reactor, the biocatalytic reaction of 2,5-DKG in 2-KLG proceeds with simultaneous regeneration of the coenzyme NADPH.
  • the sterile stirred tank with a reactor volume of 10 liters was filled with medium, which is described above for the fed-batch fermentation, and inoculated the stirred tank with the contents of two precultivation Erlenmeyer flasks of Pectobacter cypripedii HEPO1 strain.
  • the Pectobacter cypripedii strain (HEPOl) was added at 28 0 C, pH 7 or 5.6, as described above, is fermented at a dissolved oxygen concentration of 10%, yielding 137 g / l product. A portion of the culture suspension is removed from the stirred tank, filtered, and the supernatant is introduced into the enzyme reactor.
  • the reaction reaction in the enzyme reactor takes place at 25 ° C (since, as mentioned above, two reactors connected to each other, different temperatures can be used) and a pH value of 6.4, which is kept constant with ammonia ,
  • the product of this fermentation, 2,5-diketo-D-gluconic acid was added with the addition of 2 U / ml of DKR, 2 U / ml and 106 g of glucose / 1 buffer (either Na acetate or Na citrate) in the enzyme reactor in converted the immediate precursor of ascorbic acid, 2-keto-L-gluconic acid with a yield of 132 g / l.
  • the volume in the enzyme reactor is also 10 L.
  • the enzymes (DKR and GDH) and the coenzymes are retained by the ultrafiltration membrane in the enzyme reactor while the product stream (glucose, gluconic acid and 2-keto-L-gulonic acid) is withdrawn from the enzyme reactor via an ion exchange chromatography column (Column material Amberlite FPA90, Rohm and Haas) and eluted with various eluents (H 2 O, methanol and H 2 SO 4 ) to separate the product stream into its individual components, thereby making it possible to remove glucose and gluconic acid from the 2- Keto-L-gulonic acid, wherein the 2-keto-L-gulonic acid is eluted as methyl ester KLG.
  • ion exchange chromatography column Cold material Amberlite FPA90, Rohm and Haas
  • the yield of 2-keto-L-gulonic acid was 132 g / l.
  • the glucose or gluconic acid (99 g / l) are returned to the stirred tank and metabolized there by the Pectobacter cypripedii strain HEPO1 to form further 2,5-diketo-D-gluconic acid.
  • the hybrid method described forms a closed circuit in order to achieve the highest possible product yields (2-keto-L-gulonic acid) with simultaneous use of the originally added enzymes or coenzymes.

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé pour produire de l'acide ascorbique, selon lequel dans une première étape une source carbone est convertie par fermentation en acide 2,5-dicétone-D-gluconique, qui est ensuite mis en réaction de manière enzymatique avec une acide 2,5-dicétone-D-gluconique réductase provenant de Corynebacterium glutamicum pour produire de l'acide 2-céto-L-gluconique. L'objectif est d'améliorer le rendement. A cet effet, la source carbone est le glucose et la conversion du glucose en acide 2,5-dicéto-D-gluconique est directement produite par la souche Pectobacter cypripedii HEPO1 (DSMZ 12393).
PCT/AT2008/000185 2007-06-01 2008-05-29 Procédé pour produire de l'acide ascorbique comprenant l'utilisation de pectobacterium cypripedit Ceased WO2008144792A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ATA875/2007 2007-06-01
AT8752007A AT505263B1 (de) 2007-06-01 2007-06-01 Verfahren zum herstellen von ascorbinsäure

Publications (2)

Publication Number Publication Date
WO2008144792A1 true WO2008144792A1 (fr) 2008-12-04
WO2008144792A4 WO2008144792A4 (fr) 2009-03-05

Family

ID=39863017

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/AT2008/000185 Ceased WO2008144792A1 (fr) 2007-06-01 2008-05-29 Procédé pour produire de l'acide ascorbique comprenant l'utilisation de pectobacterium cypripedit

Country Status (2)

Country Link
AT (1) AT505263B1 (fr)
WO (1) WO2008144792A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021005314A1 (fr) 2019-07-11 2021-01-14 Institut Des Sciences Et Industries Du Vivant Et De L'environnement - Agroparistech Procede de purification de coenzymes a base adenosine de haut poids moleculaire par dia-filtration tangentielle

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111560334B (zh) * 2020-05-22 2021-10-29 浙江农林大学 溶磷固氮复合菌剂及其制备方法和应用

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5234819A (en) * 1980-08-14 1993-08-10 Shiongi & Co., Ltd. Method for preparing 2,5-diketo-D-gluconic acid

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DK1373480T3 (da) * 2001-04-04 2009-05-11 Genencor Int Ukoblede produktive og katabolske værtscellebaner
US20040086984A1 (en) * 2002-10-31 2004-05-06 Kado Clarence I. Microbiological method for producing ascorbic acid

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5234819A (en) * 1980-08-14 1993-08-10 Shiongi & Co., Ltd. Method for preparing 2,5-diketo-D-gluconic acid

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
ANONYMOUS: "Eintrag zu DSMZ 12393: Erwinia pyrifoliae", INTERNET ARTIKEL, 2004, XP002501122, Retrieved from the Internet <URL:http://www.dsmz.de/microorganisms/html/strains/strain.dsm012393.html> [retrieved on 20081024] *
ANONYMOUS: "Eintrag zu DSMZ 30182: Pectobacterium cypripedii (Hori 1911)", INTERNET ARTIKEL, 2004, XP002501123, Retrieved from the Internet <URL:http://www.dsmz.de/microorganisms/html/strains/strain.dsm030182.html> [retrieved on 20081024] *
PACHER, C.: "Application of NADPH-dependent 2.5-diketo-gluconic acid reductase for production of L-ascorbic acid", JOURNAL OF BIOTECHNOLOGY, vol. 118, no. Suppl1, August 2005 (2005-08-01), pages S109 - S110, XP008097843 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021005314A1 (fr) 2019-07-11 2021-01-14 Institut Des Sciences Et Industries Du Vivant Et De L'environnement - Agroparistech Procede de purification de coenzymes a base adenosine de haut poids moleculaire par dia-filtration tangentielle
FR3098416A1 (fr) * 2019-07-11 2021-01-15 Agro Paris Tech Procédé de purification de cofacteurs de haut poids moléculaire par dia-filtration tangentielle
US20220280899A1 (en) * 2019-07-11 2022-09-08 Institut des Sciences et Industries du Vivant et de I'environnement - Agroparis Tech Method for purifying high molecular weight adenosine-based coenzymes by tangential diafiltration

Also Published As

Publication number Publication date
AT505263A1 (de) 2008-12-15
WO2008144792A4 (fr) 2009-03-05
AT505263B1 (de) 2009-08-15

Similar Documents

Publication Publication Date Title
DE69931394T2 (de) Verfahren zur herstellung von ascorbinsäure-zwischenprodukten
EP2670837A1 (fr) Procédé de préparation de 2,3-butanediol par fermentation
EP1437415A1 (fr) Préparation d&#39;acide lactique d&#39;un substrat contenant des pentoses
EP2670838A1 (fr) Procédé de préparation de 2,3-butanediol par fermentation
EP2670836A1 (fr) Procédé de préparation de 2,3-butanediol par fermentation
AT503017A4 (de) Verfahren zur enantioselektiven enzymatischen reduktion von hydroxyketoverbindungen
AT505263B1 (de) Verfahren zum herstellen von ascorbinsäure
DE102016007810B4 (de) Verfahren zur Herstellung von D-Xylonat
DE3485981T2 (de) Biosynthetische 2,5-diketoglukonsaeure-reduktase, verfahren, rekombinante zellen und expressionsvektoren zu deren herstellung und ihre verwendung zur herstellung von 2-keto-l-gulonsaeure.
US5792630A (en) Cellulose-producing microorganism transformed with a gene for an enzyme involved in sucrose metabolism
JP2002520066A (ja) 固定化微生物によるマンニトールの製造方法
WO2012104233A1 (fr) Procédé de préparation de 2,3-butanediol par fermentation
EP1153120A1 (fr) Procede de preparation de l-sorbose
DE10220234B4 (de) Verfahren sowie Mikroorganismen zur mikrobiellen Herstellung von Pyruvat aus Kohlenhydraten sowie Alkoholen
CN108315375B (zh) 一种氧化型烟酰胺腺嘌呤二核苷酸磷酸的生产方法
EP4389881A1 (fr) Micro-organisme génétiquement modifié et son utilisation pour la production de d-chiro-inositol
EP4678755A1 (fr) Procédé de préparation d&#39;une solution aqueuse contenant du d-mannitol
CA1119981A (fr) Procede d&#39;obtention d&#39;acides 2,5-dicetogluconiques
WO2025012225A1 (fr) Procédé de production d&#39;une solution aqueuse contenant du l-sorbose
DE102004010786A1 (de) Mikroorganismus und Verfahren zur Herstellung von Weinsäure
GB2058775A (en) Preparation of 2,5-diketogluconic acid
WO2026013248A1 (fr) Procédé de production d&#39;une solution aqueuse contenant du d-mannitol
EP0171447B1 (fr) Procédé de préparation bactérielle de l&#39;acide gluconique
WO2024084032A1 (fr) Procédé de préparation d&#39;une solution aqueuse contenant un sel alcalin de l&#39;acide glycolique et de l&#39;acide lactique
DD275480A1 (de) Verfahren zur herstellung von zitronensaeure durch aerobe submerse kultivierung von hefen

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: 08747906

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 08747906

Country of ref document: EP

Kind code of ref document: A1