+

WO1994006925A1 - Production d'acide glyoxylique, par oxydation d'acide glycoliqu en presence de glycolate oxydase et de catalase immobilisees - Google Patents

Production d'acide glyoxylique, par oxydation d'acide glycoliqu en presence de glycolate oxydase et de catalase immobilisees Download PDF

Info

Publication number
WO1994006925A1
WO1994006925A1 PCT/US1992/008002 US9208002W WO9406925A1 WO 1994006925 A1 WO1994006925 A1 WO 1994006925A1 US 9208002 W US9208002 W US 9208002W WO 9406925 A1 WO9406925 A1 WO 9406925A1
Authority
WO
WIPO (PCT)
Prior art keywords
immobilized
catalase
glycolate oxidase
acid
glycolic acid
Prior art date
Application number
PCT/US1992/008002
Other languages
English (en)
Inventor
David Leroy Anton
Robert Dicosimo
John Edward Gavagan
Original Assignee
E.I. Du Pont De Nemours And Company
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
Priority to PCT/US1992/008002 priority Critical patent/WO1994006925A1/fr
Priority claimed from CA002144637A external-priority patent/CA2144637A1/fr
Priority to JP50803294A priority patent/JP3145711B2/ja
Priority to CZ95700A priority patent/CZ70095A3/cs
Priority to EP92920655A priority patent/EP0662142A1/fr
Priority to BR9207165A priority patent/BR9207165A/pt
Priority to AU26543/92A priority patent/AU2654392A/en
Priority to CA002144637A priority patent/CA2144637A1/fr
Application filed by E.I. Du Pont De Nemours And Company filed Critical E.I. Du Pont De Nemours And Company
Priority to HU9500950A priority patent/HU213760B/hu
Priority to NZ244529A priority patent/NZ244529A/en
Priority to ZA927518A priority patent/ZA927518B/xx
Priority claimed from NZ244592A external-priority patent/NZ244592A/en
Priority to PT100935A priority patent/PT100935A/pt
Priority to CN92112653A priority patent/CN1086263A/zh
Publication of WO1994006925A1 publication Critical patent/WO1994006925A1/fr

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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids

Definitions

  • This invention relates to an improved process for the production of glyoxylic acid by the enzyme catalyzed oxidation of glycolic acid. More specifically, the present invention relates to the use of glycolate oxidase and catalase immobilized on an insoluble support as catalyst.
  • Glycolate oxidase an enzyme commonly found in leafy green plants and mammalian cells, catalyzes the oxidation of glycolic acid to glyoxylic acid, with the concomitant production of hydrogen peroxide.
  • the addition of certain compounds, such as ethylenediamine, limited the further oxidation of the intermediate glyoxylic acid.
  • the oxidations were carried out at a pH of about 8, typically using glycolic acid concentrations of about 3-40 mM (millimolar). The optimum pH for the glycolate oxidation was reported to be 8.9. Oxalic acid (100 mM) was reported to inhibit the catalytic action of the glycolate oxidase. Similarly, K. E. Richardson and N. E. Tolbert. J. Biol. Chem.. Vol. 236, 1280-1284 (1961) showed that buffers containing tris(hydroxymethyl)aminomethane inhibited the formation of oxalic acid in the glycolate oxidase catalyzed oxidation of glycolic acid. C. O. Clagett, N. E. Tolbert and R. H. Burris.
  • This invention relates to a process for the production of glyoxylic acid (OCHCOOH) where glycolic acid (HOCH2COOH) (200 to about 2500 mM) and oxygen are reacted in an aqueous solution (pH 7 to 10), in the presence of a catalyst consisting of glycolate oxidase ((S)-2-hydroxy-acid oxidase, EC 1.1.3.15) and catalase (EC 1.11.1.6) immobilized on an insoluble support. Under optimum conditions, very high yields of glyoxylic acid are obtained at high conversion of glycolic acid, and the immobilized enzyme catalyst can be recovered and reused.
  • This invention describes the preparation and use of an immobilized enzyme catalyst for the manufacture of glyoxylic acid from glycolic acid (hydroxyacetic acid).
  • glycolic acid hydroxyacetic acid
  • high selectivities (c 99%) to glyoxylic acid have not been previously obtained, nor has the oxidation of glycolic acid been performed at concentrations of 0.20M to 2.5M.
  • U.S.S.N. 07/422,011 filed Oct.
  • a previously-reported use of soluble enzymes as catalysts poses several problems: catalyst recovery for reuse is not easily performed, catalyst stability is not as good as can be obtained with immobilized enzyme systems, and soluble enzymes are not stable to the sparging of the reaction mixture with oxygen (required to increase the rate of oxygen dissolution and, thus, reaction rate).
  • a catalyst preparation has now been developed which involves the simultaneous immobilization of the two enzymes; i.e., glycolate oxidase (e.g., from spinach or beet leaves, isolated or obtained from commercial sources) and catalase (e.g., from Aspergillus niger. Aspergillus nidulans.
  • the immobilized catalyst is easily recovered from the reaction mixture at the conclusion of the reaction for reuse, whereas the soluble enzyme is only recovered with great difficulty and loss of activity; 2) the immobilized catalyst is more stable than the soluble enzyme, both for the number of catalyst turnovers obtained versus the soluble enzyme, as well as for recovered enzyme activity at the conclusion of a reaction or after prolonged storage in aqueous buffer; and 3) most importantly, the immobilized catalyst is stable to reaction conditions where oxygen is sparged into the reaction mixture to increase the rate of oxygen dissolution and reaction rate, where under similar reaction conditions the soluble glycolate oxidase is rapidly denatured.
  • the immobilization of enzymes can be performed using a variety of techniques, including: (1) binding of the enzyme to a carrier or support, via covalent attachment, physical adsorption, electrostatic binding, or affinity binding, (2) crosslinking with bifunctional or multifunctional reagents, (3) entrapment in gel matrices, polymers, emulsions, or some form of membrane, and (4) a combination of any of these methods.
  • the stability of immobilized glycolate oxidase in aqueous buffers is much greater than the soluble enzyme (approaching the stability of ammonium sulfate-precipitated enzyme).
  • Recovery and reuse of the co-immobilized catalyst was easily performed by simply filtering the catalyst away from the reaction mixture and recycling it to fresh reaction mixture; in this manner for immobilized glycolate oxidase the number of turnovers (i.e., the number of substrate molecules that are converted to product molecules per catalyst molecule before inactivation of the enzyme) as high as 10
  • the immobilized glycolate oxidase used in the reaction should be present in an effective concentration, usually a concentration of about 0.001 to about 10.0 IU/mL, preferably about 0.1 to about 4 IU/mL.
  • An IU IU/mL
  • This method is also used to assay the activity of recovered or recycled glycolate oxidase.
  • the pH of the reaction solution should be between 7 and 10, preferably between 8.0 and 9.5.
  • the pH can be maintained by a buffer, since enzyme activity varies with pH.
  • the pH of the reaction decreases slightly as the reaction proceeds, so it is often useful to start the reaction near the high end of the maximum enzyme activity pH range, about
  • an amine buffer capable of complexing the glyoxylic acid (by forming an imine which is more stable to chemical or enzymatic oxidation) is employed along with catalase to maximize product selectivity.
  • Ethylenediamine, or less preferably, tris(hydroxymethyl)methylamine (hereinafter TRIS), piperazine, or glycylglycine improved the yield of glyoxylic acid.
  • TRIS tris(hydroxymethyl)methylamine
  • piperazine or glycylglycine
  • These amines are used in a molar ratio of amine/glycolic acid (starting amount) of 1.0 to 3.0, preferably 1.05 to 1.33.
  • the exact value may be adjusted to obtain the desired pH.
  • very basic amines used at high amine to glycolic acid ratios it may be necessary to adjust the pH, as by adding acid, for example hydrochloric or sulfuric acids.
  • acid for example hydrochloric or sulfuric acids.
  • less basic amines such as TRIS it may be necessary to add a base to maintain the desired pH.
  • the concentration of immobilized catalase should be 50 to 100,000 IU/mL, preferably 350 to 14,000 IU/mL. It is preferred that the enzymes be co-immobilized to limit the amount of catalyst added to the reaction, and that the catalase and glycolate oxidase concentrations be adjusted within the above ranges so that the ratio (measured in IU for each) of catalase:glycolate oxidase is at least about 250:1.
  • Flavinmononucleotide (FMN) is an optional added ingredient, used at a concentration of 0.0 to 2.0 mM, preferably 0.01 to 0.2 mM.
  • the reaction rate is at least partially controlled by the rate at which oxygen can be dissolved into the aqueous medium.
  • Oxygen can be added to the reaction as the oxygen in air, but it is preferred to use a relatively pure form of oxygen, and to use elevated pressures. Although no upper limit of oxygen pressure is known, oxygen pressures up to 50 atmospheres may be used, and an upper limit of 15 atmospheres is preferred. Sparging (bubbling) oxygen through the reaction mixture is necessary to maintain a high oxygen dissolution (and hence reaction) rate. Oxygen is sparged through the reaction mixture at a rate of 0.05 to 5 volumes of oxygen (measured at atmospheric pressure) per volume of reaction mixture per minute (vol/vol-min), and preferably between 0.2 and 2 vol/vol-min.
  • reaction temperature is an important variable, in that it affects reaction rate and the stability of the enzymes.
  • a reaction temperature of 0°C to 40°C may be used, but the preferred reaction temperature range is from 5°C to 15°C. Operating in the preferred temperature range maximizes recovered enzyme activity at the end of the reaction.
  • amine buffer is most conveniently removed by use of an ion exchange resin.
  • Suitable acidic cationic exchange resins include "AMBERLITE” CG120 or “AMBERLITE” IR120 (Rohm & Haas Co.), and “DOWEX” 50 (Dow Chemical Co.). The amine may then be recovered and subsequently recycled by treatment of the resin with strong base.
  • glyoxylic acid is useful in the preparation of vanillin and ethylvanillin, as well as being used in ion exchange resins and as an acid catalyst in the pharmaceutical industry (Ullmanns). It is usually sold as a 50% (weight percent) aqueous solution. It is also to be understood that reference to glyoxylic acid in this application can also mean the glyoxylate anion, especially when the glyoxylic acid is present in a solution whose pH is greater than about 2.3.
  • Glycolate oxidase from spinach was purified using selective ammonium sulfate fractionation followed by batch adsorption of the extract using DEAE cellulose. The latter procedure resulted in the adsorption of virtually all plant proteins except glycolate oxidase. All steps in the purification were performed at 4°C unless otherwise stated. At 25°C, two bushels (16 kg) of fresh spinach were chopped into fine particles using a Fitz Mill grinder fitted with a 0.5 inch mesh screen. The liquid fraction (ca. 6 L) of the resulting pulp was isolated by squeezing through 4 layers of cheesecloth; alternatively, a juice extractor (Nitantonio) may be used.
  • the protein pellet was dissolved in approx. 200 mL of 20 mM bicine buffer (pH 8.0). Using Spectropor 2 dialysis tubing (12,000-14,000
  • the protein was dialysed for 16 hrs. vs. 4 L of 20 mM bicine (pH 8.0) containing 2 mM FMN.
  • the conductivity of the protein solution was measured relative to the conductivity of fresh bicine buffer using a conductivity meter, and if the readings were not equivalent, the protein solution was dialysed an additional 4 hrs, then tested as before.
  • the dialysed protein solution (approx. 250 ml) was stirred in a beaker using either a magnetic stir bar or overhead stirrer, then 25 g of pre-swollen DEAE cellulose (Sigma) (Kerr, M. W., Groves, P.. Phytochemistry. Vol. 14, 359-362 (1975)) added and the resulting mixture incubated for 10 minutes. Protein binding to the resin was monitored by following the decrease in protein concentration of the solution using the
  • a 200 mL portion of the yeast suspension was transferred to a 400 mL capacity
  • a 50-mL portion of the dialysed protein was loaded onto a radial flow chromatography column (Sepragen) packed with 100 mL of Q Sepharose fast flow ion exchange resin (Pharmacia), and the unbound protein eluted with 20 mM TRIS (pH 7.5) at 10 mL/min. Protein elution was monitored using a flow cell fitted with a 280 nm filter (LKB) linked to a chart recorder (LKB); 10-15 mL column fractions were collected using an LKB fraction collector.
  • LLB 280 nm filter
  • LLB chart recorder
  • This purification method has also been used to purify catalase from Aspergillus nidulans and Aspergillus niger.
  • Glycolate oxidase immobilized on oxirane acrylic beads was assayed by accurately weighing ca. 5-10 mg of the treated beads into a 3-mL quartz cuvette containing a magnetic stirring bar, then 2.0 mL of a solution which was 0.12 mM in
  • Samples for analysis were prepared by mixing 0.100 mL of the reaction mixture with 0.300 mL of 0.1 N H2SO4, then filtering the resulting solution through a Millipore Ultrafree MC filter unit (10,000 mw cutoff).
  • Analyses for glycolic acid, glyoxylic acid, oxalic acid and formic acid were performed by high performance liquid chromatography (HPLC) on a Bio-Rad Aminex HPX-87H column (300 x 7.8 mm) at 40°C, using as solvent an aqueous solution of H2SO4(0.01 N) and l-hydroxyethane-l,l-diphosphonic acid (0.1 mM) at 1.0 mL/minute.
  • the instrument was a Waters 840 HPLC system with Model 510 pumps, a 712 WISP autosampler, and, in sequence, a 490E UN detector and 410 differential refractometer. UN analysis was performed at 210 nm. The retention times for oxalic acid, glyoxylic acid, glycolic acid, formic acid, and propionic acid (internal standard) were 4.29, 6.09, 7.77, 8.79, and 11.41 minutes, respectively.
  • PION Poly(ethyleneimine)
  • poly(ethyl-eneimine) on silica gel benzylated poly(ethyl-eneimine) on silica gel
  • Bio-Rex 70 CH Sepharose 4B
  • XAD-4, XAD-8 Phenyl Agarose
  • Eupergit C Eupergit C-250L
  • Eupergit C-30 ⁇ were all obtained from commercial sources.
  • PAN-500 poly(acrylamide-co-N-acryloxysuccinimide) gel crosslinked with triethylenetetramine
  • oxirane acrylic beads Eupergit C
  • a solution containing 50 mM bicine buffer (pH 8.0) and 0.02 mM flavin mononucleotide 50 mM bicine buffer (pH 8.0) and 0.02 mM flavin mononucleotide
  • the oxirane acrylic beads were then suspended in the buffer by swirling the contents of the flask. After the beads had settled to the bottom of the flask, the fines which floated to the top of the mixture were removed by pipet, along with as much of the supernatant which could be removed without disturbing the settled beads. This washing procedure was repeated a second time.
  • a 10-mL mixture containing 100 mg (715,000 IU) of ammonium sulfate-precipitated Aspergillus niger catalase (Sigma C-3515) was centrifuged at 15,000 rpm for 10 minutes (Sorvall SS-34 rotor). The supernatant was discarded and the pellet dissolved in the buffer containing the glycolate oxidase. This enzyme solution was then added to the flask containing the washed oxirane acrylic beads, and the final volume adjusted to 125 mL with additional buffer. The resulting mixture was transferred to a 250-mL polypropylene bottle, which was capped and placed on a bottle roller at 4-5 rpm for 16 hours at 15°C.
  • the mixture was then poured into a chromatography column equipped with a fritted bed support, allowed to drain, and the immobilized enzymes were washed three times with 30 mL of the bicine/FMN buffer and stored at 5°C in this same buffer.
  • the co-immobilized enzyme catalyst had 7.2 IU of glycolate oxidase activity /gram Eupergit C and 5680 IU of catalase activity/gram Eupergit C.
  • Example 3 Relative Stability of Soluble and Immobilized Glycolate Oxidase The stability of unimmobilized (soluble) glycolate oxidase versus glycolate oxidase immobilized on oxirane acrylic beads (Eupergit C) was measured by storing either form of the enzyme at 4°C in a buffered (pH 8.0) solution containing 2.0 mM flavin mononucleotide, then monitoring the enzyme activity with time. Additionally, the stability of the enzyme precipitated in 3.2 M ammonium sulfate, 2.0 mM flavin mononucleotide, and stored under similar conditions was also monitored. Enzyme
  • Example 4 Sparged Co-Immobilized Enzyme Reaction Into a 2.5-cm ID x 20 cm glass column equipped with a 20-mm polyethylene bed support was placed 10 mL of a solution containing glycolic acid (0.25 M), ethylenediamine (0.33 M), propionic acid (0.075 M, HPLC internal standard), and flavin monunucleotide (0.2 mM). The column and its contents were cooled to 15°C, then 2.5 IU of spinach glycolate oxidase and 27,000 IU of Aspergillus niger catalase (co-immobilized on Eupergit C) were added to the solution.
  • Oxygen was then passed through the porous bed support and bubbled through the reaction mixture at a rate of 10 mL/min.
  • the reaction was monitored by taking a 100 mL aliquot of the reaction mixture at regular intervals, mixing the aliquot with 300 mL of 0.1 N sulfuric acid to quench the reaction, filtering the aliquot and analyzing by HPLC. After 5.5 hours, the yields of glyoxylic acid, oxalic acid, and formic acid were 98%, 2%, and 0%, respectively, with complete conversion of glycolic acid.
  • the final activities of glycolate oxidase and catalase were 95% and 65% of their initial values.
  • Example 4 The reaction described in Example 4 was repeated, except that the same amounts of soluble, unimmobihzed glycolate oxidase and catalase were added to the reaction mixture. After 4 hours, the yields of glyoxylic acid, oxalic acid, and formic acid were 43%, 0%, and 0%, respectively, with a 46% conversion of glycolic acid. The final activities of glycolate oxidase and catalase were ⁇ 2% and 82% of their initial values, respectively, and no further reaction was observed at longer reaction times.
  • Example 4 The reaction in Example 4 was repeated using 10 mL of a solution containing glycolic acid (0.75 M), ethylenediamine (0.86 M), propionic acid
  • the yields of glyoxylic acid, oxalic acid, and formic acid were 93%, 0%, and 0.3%, respectively, with a 94% conversion of glycolic acid.
  • the final activities of glycolate oxidase and catalase were 48% and 69% of their initial values.
  • a 300-mL EZE-Seal stirred autoclave (Autoclave Engineers) was charged with 100 mL of a solution containing glycolic acid (0.75 M), ethylenediamine (0.86 M, pH 9.0), propionic acid (0.075 M, HPLC internal standard), and flavin monunucleotide (0.01 mM), and the solution cooled to 15°C.
  • To the autoclave was then added 89 IU of spinach glycolate oxidase and 72,600 IU of Aspergillus niger catalase co-immobilized on ca.28 g of Eupergit C. The resulting mixture was stirred at 500 rpm and 15°C.
  • the immobilized enzyme catalyst was recovered from the reaction described in Example 7 by filtering the reaction mixture through a 2.5-cm ID x 20 cm glass column equipped with a 20-mm polyethylene bed support. The remaining liquid adsorbed on the catalyst was removed by briefly passing a stream of nitrogen through the column, then the catalyst was resuspended in 100 mL of a fresh 15°C solution containing glycolic acid (0.75 M), ethylenediamine (0.86 M), propionic acid (0.075 M HPLC internal standard), and flavin mononucleotide (0.01 mM). The 300-mL autoclave reactor was again charged with this reaction mixture, and the reaction repeated. This catalyst recovery procedure was performed for 10 consecutive batch reactions, and the reaction time, the recovery of glycolate oxidase (G.O.) and catalase activity, and yield of glyoxylic acid are listed in the table below.
  • a 300-mL EZE-Seal stirred autoclave (Autoclave Engineers) was charged with 100 mL of a solution containing glycolic acid (0.75 M), ethylenediamine (0.86 M, pH 9.0), propionic acid (0.075 M, HPLC internal standard), and flavin mononucleotide (0.01 mM), and the solution cooled to 15°C.
  • To the autoclave was then added 41 IU of spinach glycolate oxidase and 42,800 IU of Aspergillus niger catalase co-immobilized on ca. 15 g of Eupergit C.
  • the resulting mixture was stirred at 400 rpm and 15°C under 35, 70, 105, or 140 psig (242, 483, 724 or 965 kPa) of oxygen, while bubbling oxygen through the mixture at 50 mL/min.
  • the reaction was monitored by taking a 100 mL aliquot of the reaction mixture at regular intervals, mixing the aliquot with 300 mL of 0.1 N sulfuric acid to quench the reaction, filtering the aliquot and analyzing by HPLC.
  • the rates for reactions run under 35, 70, 105, or 140 psig (242, 483, 724 or 965 kPa) of oxygen were 0.48, 0.54, 0.53, and 0.57 mmol/min of glycolic acid, respectively.
  • Example 9 The reactions in Example 9 were repeated in a stirred autoclave reactor, except that no oxygen was bubbled through the reaction mixtures.
  • Example 10 Enzymatic oxidation of Glycolic Acid Using Permeabilized Bakers Yeast with Immobilized Glycolate Oxidase
  • the procedures described in Examples 7 and 8 were repeated, except that 50 IU of spinach glycolate oxidase immobilized on ca. 15 g of Eupergit C, and 4.0 g of fresh Saccharomyces cerevisiae (Bakers yeast, Red Star brand, Universal Foods) which had been permeabilized with isopropanol and contained 100,000 IU of catalase activity, were used as catalyst.
  • the reaction mixture was stirred at 400 rpm and 15°C under 70 psig (483 kPa) of oxygen, while bubbling oxygen through the mixture at 20 mL/min.
  • Six consecutive batch reactions were run, and the reaction time, the recovery of glycolate oxidase (G.O.) and catalase activity, and yield of glyoxylic acid are listed in the table below.
  • Example 7 The procedure described in Example 7 was repeated using 80, 60, 40, or 20 IU of spinach glycolate oxidase and 1,400,000, 1,000,000, 70,000, or 35,000 IU of Aspergillus niger catalase co-immobilized, respectively, on Eupergit C.
  • the reaction mixture was stirred at 400 rpm and 15°C under 70 psig of oxygen, while sparging oxygen through the mixture at 20 mL/min.
  • the rates of glycolic acid oxidation obtained using different concentrations of glycolate oxidase is listed in the table below.

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)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

Procédé de production d'acide glyoxylique consistant à faire réagir de l'acide glycolique avec de l'oxygène dans une solution aqueuse en présence d'un tampon d'amine capable de former un produit d'addition chimique avec de l'acide glyoxylique, et de glycolate et de catalase immobilisées ou co-immobilisées sur un suport insoluble. Ladite réaction est effectuée à un pH de 7 à 10, de préférence 8 à 9,5, à une concentration initiale d'acide glycolique de 200 à 2500 mM, à une concentration d'amine, dans laquelle le rapport molaire initial de l'amine à l'acide glycolique est de l'ordre de 1,0 à 3,0, à une concentration de catalase immobilisée de 50 à 100.000 IU/mL, de préférence 350 à 14.000 IU/mL, à une pression d'oxygène allant jusqu'à 50 atmosphères, de préférence 15 atmosphères, à une concentration de glycolate oxydase immobilisée d'environ 0,01 à 10 IU/mL, de préférence environ 0,1 à 4 IU/mL, et à une température de 0 °C à 40 °C, de préférence de 5 °C à 15 °C. Des supports d'immobilisation insolubles préférés sont Eupergit C-250L et Eupergit C (bourrelets acryliques d'oxirane).
PCT/US1992/008002 1992-09-18 1992-09-18 Production d'acide glyoxylique, par oxydation d'acide glycoliqu en presence de glycolate oxydase et de catalase immobilisees WO1994006925A1 (fr)

Priority Applications (12)

Application Number Priority Date Filing Date Title
AU26543/92A AU2654392A (en) 1992-09-18 1992-09-18 Production of glyoxylic acid by oxidizing glycolic acid in the presence of immobilized glycolate oxidase and catalase
JP50803294A JP3145711B2 (ja) 1992-09-18 1992-09-18 固定化グリコレートオキシダーゼ及びカタラーゼの存在下における，グリコール酸の酸化によるグリオキシル酸の製造
CZ95700A CZ70095A3 (en) 1992-09-18 1992-09-18 Preparation of glyoxylic acid by oxidizing glycollic acid in the presence of immobilized glycolate oxidase and catalase
EP92920655A EP0662142A1 (fr) 1992-09-18 1992-09-18 Production d'acide glyoxylique, par oxydation d'acide glycoliqu en presence de glycolate oxydase et de catalase immobilisees
BR9207165A BR9207165A (pt) 1992-09-18 1992-09-18 Processo para a produção de ácido glioxílico
CA002144637A CA2144637A1 (fr) 1992-09-18 1992-09-18 Preparation d'acide glyoxylique par oxydation de l'acide glycolique en presence de glycolate oxydase et catalase immobilisees
HU9500950A HU213760B (en) 1992-09-18 1992-09-18 Process for the production of glyoxylic acid by enzyme catalysed oxidation of glycolic acid
PCT/US1992/008002 WO1994006925A1 (fr) 1992-09-18 1992-09-18 Production d'acide glyoxylique, par oxydation d'acide glycoliqu en presence de glycolate oxydase et de catalase immobilisees
NZ244529A NZ244529A (en) 1992-09-18 1992-09-28 Production of glyoxylic acid comprising contacting glycolic acid, glycolate oxidase and catalase in an aqueous solution
ZA927518A ZA927518B (en) 1992-09-18 1992-09-30 Production of glyoxylic acid by oxidizing glycolic acid in the presence of immobilized glycolate oxidase and catalase
PT100935A PT100935A (pt) 1992-09-18 1992-10-07 Processo para a preparacao de acido glioxilico por oxidacao de acido glicolico na presenca de glicolato oxidase e catalase imobilizadas
CN92112653A CN1086263A (zh) 1992-09-18 1992-10-26 二羟乙酸的生产方法

Applications Claiming Priority (9)

Application Number Priority Date Filing Date Title
CA002144637A CA2144637A1 (fr) 1992-09-18 1992-09-18 Preparation d'acide glyoxylique par oxydation de l'acide glycolique en presence de glycolate oxydase et catalase immobilisees
HU9500950A HU213760B (en) 1992-09-18 1992-09-18 Process for the production of glyoxylic acid by enzyme catalysed oxidation of glycolic acid
PCT/US1992/008002 WO1994006925A1 (fr) 1992-09-18 1992-09-18 Production d'acide glyoxylique, par oxydation d'acide glycoliqu en presence de glycolate oxydase et de catalase immobilisees
CS7009592 1992-09-18
NZ244529A NZ244529A (en) 1992-09-18 1992-09-28 Production of glyoxylic acid comprising contacting glycolic acid, glycolate oxidase and catalase in an aqueous solution
ZA927518A ZA927518B (en) 1992-09-18 1992-09-30 Production of glyoxylic acid by oxidizing glycolic acid in the presence of immobilized glycolate oxidase and catalase
NZ244592A NZ244592A (en) 1991-10-04 1992-10-02 Improving the selectivity of delignification of chemical pulp using an
PT100935A PT100935A (pt) 1992-09-18 1992-10-07 Processo para a preparacao de acido glioxilico por oxidacao de acido glicolico na presenca de glicolato oxidase e catalase imobilizadas
CN92112653A CN1086263A (zh) 1992-09-18 1992-10-26 二羟乙酸的生产方法

Publications (1)

Publication Number Publication Date
WO1994006925A1 true WO1994006925A1 (fr) 1994-03-31

Family

ID=38595975

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1992/008002 WO1994006925A1 (fr) 1992-09-18 1992-09-18 Production d'acide glyoxylique, par oxydation d'acide glycoliqu en presence de glycolate oxydase et de catalase immobilisees

Country Status (10)

Country Link
EP (1) EP0662142A1 (fr)
JP (1) JP3145711B2 (fr)
CN (1) CN1086263A (fr)
AU (1) AU2654392A (fr)
CZ (1) CZ70095A3 (fr)
HU (1) HU213760B (fr)
NZ (1) NZ244529A (fr)
PT (1) PT100935A (fr)
WO (1) WO1994006925A1 (fr)
ZA (1) ZA927518B (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6323011B1 (en) 1996-03-23 2001-11-27 Institute Of Food Research Production of vanillin
US11806335B2 (en) 2019-11-01 2023-11-07 Lilac Therapeutics, Inc. Heterocyclic carboxylate compounds as glycolate oxidase inhibitors

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109988784B (zh) * 2019-04-16 2021-02-02 台州学院 一种固定化甘醇酸氧化酶催化合成丙酮酸的方法

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991005868A1 (fr) * 1989-10-16 1991-05-02 E.I. Du Pont De Nemours And Company Production d'acide glyoxylique par oxydation enzymatique d'acide glycolique

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991005868A1 (fr) * 1989-10-16 1991-05-02 E.I. Du Pont De Nemours And Company Production d'acide glyoxylique par oxydation enzymatique d'acide glycolique

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6323011B1 (en) 1996-03-23 2001-11-27 Institute Of Food Research Production of vanillin
US6664088B2 (en) 1996-03-23 2003-12-16 Plant Bioscience Limited Production of vanillin
US11806335B2 (en) 2019-11-01 2023-11-07 Lilac Therapeutics, Inc. Heterocyclic carboxylate compounds as glycolate oxidase inhibitors
US12168002B2 (en) 2019-11-01 2024-12-17 Lilac Therapeutics, Inc. Heterocyclic carboxylate compounds as glycolate oxidase inhibitors

Also Published As

Publication number Publication date
CN1086263A (zh) 1994-05-04
EP0662142A1 (fr) 1995-07-12
HU213760B (en) 1997-09-29
NZ244529A (en) 1995-06-27
ZA927518B (en) 1994-03-30
PT100935A (pt) 1994-06-30
HU9500950D0 (en) 1995-05-29
JP3145711B2 (ja) 2001-03-12
JPH08501218A (ja) 1996-02-13
CZ70095A3 (en) 1995-11-15
AU2654392A (en) 1994-04-12
HUT72915A (en) 1996-06-28

Similar Documents

Publication Publication Date Title
US4221869A (en) Enzymatic synthesis of L-carnitine
AU686182B2 (en) Process for the preparation of pyruvic acid
Huwig et al. Laboratory procedures for producing 2-keto-D-glucose, 2-keto-D-xylose and 5-keto-D-fructose from D-glucose, D-xylose and L-sorbose with immobilized pyranose oxidase of Peniophora gigantea
Seip et al. Biocatalytic production of glyoxylic acid
Seip et al. Glyoxylic acid production using immobilized glycolate oxidase and catalase
US5439813A (en) Production of glyoxylic acid with glycolate oxidase and catalase immobilized on oxirane acrylic beads
EP0056038B1 (fr) Procede a hydrate de carbone
EP0496799B1 (fr) Production d'acide glyoxylique par oxydation enzymatique d'acide glycolique
WO1994006925A1 (fr) Production d'acide glyoxylique, par oxydation d'acide glycoliqu en presence de glycolate oxydase et de catalase immobilisees
US6500649B2 (en) Process for the conversion of organic materials, particularly saccharide materials, comprising an enzymatic oxidation step in the presence of ruthenium or palladium
US5219745A (en) Production of glyoxylic acid from glycolic acid
CA2144637A1 (fr) Preparation d'acide glyoxylique par oxydation de l'acide glycolique en presence de glycolate oxydase et catalase immobilisees
EP0621900B1 (fr) Oxydation d'acide glycolique en acide glycoxylique a l'aide d'une cellule transformee microbienne utilisee comme catalyseur
US5221621A (en) Production of glyoxylic acid from glycolic acid
US5149646A (en) Process for isolating galactose oxidase
JP3709317B2 (ja) 光学活性シアノヒドリンの合成方法
US5262314A (en) Enzymatic oxidation of glycolic acid in the presence of non-enzymatic catalyst for decomposing hydrogen peroxide
US5538875A (en) Process for the preparation of pyruvic acid using permeabilized transformants of H. polymorha and P. pastoris which express glycolate oxidase and catalase
Schmidt et al. Comparison of chemical and biochemical reduction methods for the synthesis of (R)-2-hydroxy-4-phenylbutyric acid
CA1150656A (fr) Methode de production de carbohydrates
SU1041567A1 (ru) Способ получени водорастворимого иммобилизованного протеолитического комплекса
JP2006521102A (ja) 組合わされた補因子依存型酵素反応系

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU BB BG BR CA CS FI HU JP KP KR LK MG MN MW NO PL RO RU SD

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LU MC NL SE BF BJ CF CG CI CM GA GN ML MR SN TD TG

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 1992920655

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2144637

Country of ref document: CA

WWE Wipo information: entry into national phase

Ref document number: PV1995-700

Country of ref document: CZ

WWP Wipo information: published in national office

Ref document number: 1992920655

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: PV1995-700

Country of ref document: CZ

WWW Wipo information: withdrawn in national office

Ref document number: 1992920655

Country of ref document: EP

WWR Wipo information: refused in national office

Ref document number: PV1995-700

Country of ref document: CZ

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载