WO2003100043A1 - Nouvelle (d)-2-hydroxy acide oxydase derivee d'un micro-organisme et procede biochimique permettant de produire de l'acide glycoxylique - Google Patents
Nouvelle (d)-2-hydroxy acide oxydase derivee d'un micro-organisme et procede biochimique permettant de produire de l'acide glycoxylique Download PDFInfo
- Publication number
- WO2003100043A1 WO2003100043A1 PCT/JP2003/006367 JP0306367W WO03100043A1 WO 2003100043 A1 WO2003100043 A1 WO 2003100043A1 JP 0306367 W JP0306367 W JP 0306367W WO 03100043 A1 WO03100043 A1 WO 03100043A1
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- WIPO (PCT)
- Prior art keywords
- acid
- oxidase
- enzyme
- dalioxylic
- activity
- Prior art date
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0006—Oxidoreductases (1.) acting on CH-OH groups as donors (1.1)
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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
- C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
- C12P7/58—Aldonic, ketoaldonic or saccharic acids
Definitions
- the present invention relates to a novel (D) -2-hydroxyacid oxidase derived from a microorganism having the ability to convert glycolic acid to dalioxylic acid.
- the present invention also provides a biochemical production of daloxylic acid, which comprises oxidizing glycolic acid with the above-mentioned oxidase and Z or a microorganism producing the oxidase, or a processed product thereof to convert the acid to dalioxylic acid.
- daloxylic acid which comprises oxidizing glycolic acid with the above-mentioned oxidase and Z or a microorganism producing the oxidase, or a processed product thereof to convert the acid to dalioxylic acid.
- Dalioxylic acid is used as a raw material for the synthesis of vanillin, ethyl vanillin, etc., and is also a useful compound as an intermediate in the synthesis of agricultural chemicals and pharmaceuticals.
- chemical methods such as nitric acid oxidation of dalyoxal have been known as methods for producing daroxylic acid, and at present most of daloxylic acid is produced by these chemical methods.
- chemical methods such as nitric acid nitric acid oxidation easily produce by-products such as organic acids other than dalioxylic acid, which adversely affects the quality of the produced dalioxylic acid and removes these by-products This requires a complicated process.
- there is a problem in treating a large amount of salt waste generated in the neutralization process such as nitric acid used in large quantities.
- dalicholate oxidase derived from spinach is used to convert dalicholate to dalioxylate.
- Japanese Patent Application Laid-Open Publication No. Hei 7-520985, Japanese Patent Application Publication No. Hei 8-5108159 are known.
- Glycolic acid oxidase has been conventionally recognized mainly in green plants.
- plant-derived glycolate oxidase such as spinach has a relatively high activity on dalioxylic acid.
- dalioxylic acid is further oxidized and oxidized.
- By-products such as acids are formed.
- oxidase is used in the oxidation reaction of dalicholate to dalioxylic acid
- oxygen may be used in addition to the substrate glycolic acid, and oxidized coenzymes such as NAD or NADP, such as dehydrogenase
- oxidized coenzymes such as NAD or NADP, such as dehydrogenase
- the use of oxidase for the oxidation reaction of glycolic acid to dalioxylic acid is an industrially advantageous method.
- no oxidase-type enzyme showing high activity and selectivity for dalicholate has been found at present. Disclosure of the invention
- the present invention provides an oxidase derived from a microorganism having an activity of converting glycolic acid to dalioxylic acid, and an efficient method for producing daloxylic acid using an oxidase derived from the microorganism or a microorganism producing the enzyme.
- the purpose is to provide.
- the present inventors have conducted intensive studies to develop an efficient method for producing dalioxylic acid, and as a result, found a microorganism having high oxidase activity against dalicholate in soil and having the activity from the microorganism.
- the present invention has been completed by isolating and purifying the enzyme and conducting detailed studies. That is, the present invention provides a novel oxidase that converts dalicholate into dalioxylic acid, and the action of cells and Z or treated cells of microorganisms producing the enzyme on glycolic acid to convert it into dalioxylic acid.
- a method for producing dalioxylic acid comprising accumulating and collecting this.
- FIG. 1 is a diagram showing the principle of a method for measuring the activity of an oxidase reaction.
- FIG. 2 is a diagram showing the temperature stability of the enzyme of the present invention.
- FIG. 3 is a diagram showing the optimal pH of the reaction of the enzyme of the present invention.
- Awake is 1M phosphate buffer as buffer, mouth is 0.lMTris_HC
- FIG. 4 is a diagram showing the pH stability of the enzyme of the present invention.
- the enzyme for converting glycolic acid to dalioxylic acid of the present invention has the following physicochemical properties (1) and (2):
- the enzyme of the present invention includes the following physicochemical properties (3):
- enzymes of the present invention include the following physicochemical properties:
- the microorganism used as a source of the (D) -2-hydroxyacid oxidase of the present invention is not particularly limited, and examples thereof include bacteria belonging to the genus Arthrobacter (Arthrobacter).
- Arthrobactors p. KNK-GA1 deposited by the National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary, Japan Tsukuba, Ibaraki, Japan) 1 Higashi 1-chome 1 Chuo No. 6 (Zip code 305-8566), Deposit date: March 7, 2002, Accession number: FERM BP-8375).
- KNK-GA 1 strain (deposited organization National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary, Address: 1-1, Higashi 1-chome, Tsukuba, Ibaraki, Japan; Deposit date 2002 On March 7, (D) -2-hydroxy oxidase derived from accession number FERM BP-8375) can be isolated and purified, for example, as follows. First, the KNK-GA1 strain (deposited organization, National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary, Tsukuba East 1-chome, Ibaraki, Japan, Chuo No. 6 (zip code 305-8566), date of deposit) On March 7, 2003, accession number FERM BP-8375) is cultured in an appropriate medium.
- the medium used in this case is not particularly limited, but for example, 5 g of glycolic acid, 1 g of yeast extract, 3.5 g of potassium dihydrogen phosphate, 6.5 g of ammonium hydrogen phosphate, 7.5 g of magnesium sulfate 0.5 g of hydrate, 0.02 g of zinc sulfate heptahydrate 0.03 g of ferrous sulfate heptahydrate 0.002 g of copper sulfate pentahydrate 0.002 g of calcium chloride dihydrate 0
- a medium (pH 7) having a composition of lg and 0.3 g of sodium chloride (both per liter) can be preferably used.
- the cells were collected from the resulting culture by centrifugation, suspended in 0.05M phosphate buffer (pH7), disrupted with Dynomill (Dyno-Mi11), and centrifuged. Remove the supernatant to obtain the supernatant (cell-free extract). Prominin sulfate is added to the resulting supernatant, and the resulting insoluble matter is removed by centrifugation to remove nucleic acids.
- the enzyme having the desired activity can be obtained from the resulting prominin-treated solution by various chromatographic methods such as salting out (ammonium sulfate), ion exchange chromatography, hydrophobic chromatography, and adsorption chromatography. Can be further purified.
- (D) — 2-Hydroxy acid oxidase of the present invention is obtained from the above KNK-GA1 strain (deposited organization, National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary, 1st address, 1-1-1, Higashi, Tsukuba, Ibaraki, Japan) 1 Has almost the same properties as (D) -2-hydroxy oxidase derived from Chuo No. 6 (Zip code 305-8566), Deposit date: March 7, 2002, Accession number FER M BP-8375) As long as it is an enzyme, it may be a natural enzyme or a recombinant enzyme.
- the recombinant enzyme is the KNK-GA1 strain (deposited by the National Institute of Advanced Industrial Science and Technology (AIST), Patent Organism Depositary, 1-1-1 Higashi, Tsukuba, Ibaraki, Japan 1 Chuo No. 6 (zip code 305-5-8566 ), Deposit date: March 7, 2002, Substitution or deletion of one or several amino acids in the amino acid sequence of (D)-2-hydroxyoxidase derived from accession number FERM BP-8375) It can be made by losing, inserting or adding.
- KNK-GA1 strain (Deposited organization: National Institute of Advanced Industrial Science and Technology (AIST), Patent Organism Depositary: Yuichi, Address: Tsukuba East 1-chome, Ibaraki, Japan 1 Chuo No. 6 (zip code 305-8566) Deposit date: March 7, 2002, accession number FERM BP-8 375) may be mutated, or (D) -2-hydroxy oxidase obtained from other natural or unnatural microorganisms .
- the activity of the oxidase reaction is measured by adding 0.1 ml of enzyme solution to 0.9 ml of 10 OmM phosphate buffer (pH 7) having the following composition, and adding a wavelength of 555 at 30 ° C. This is done by measuring the increase in nm absorbance.
- the enzyme activity of producing of H 2 ⁇ 2 l ino 1 is defined as 1 un it to 1 minute.
- 4-aminoaminopyrine (hereinafter 4-AA) 0.67 mM
- glycolic acid and dalioxylic acid can be analyzed by high performance liquid chromatography.
- Analysis by high performance liquid chromatography is performed, for example, using a Bio-Rad Aminex HPX-87H (7.8 mm x 300 mm) column, using a 5 mM aqueous solution of H 2 SO 4 as a solvent at a flow rate of 4 m 1 / min. .
- Detection is performed by measuring the absorbance at 23 OmM. Under these conditions, glycolic acid is eluted 20 minutes after initiation and dalioxylic acid is eluted 15 minutes after initiation.
- Known glycolic acid oxidases are active on 2-hydroxy acids in addition to glycolic acid. It is active in 2-hydroxyisopropanoic acid and hardly active in D-lactic acid. Due to its stereospecificity, it is now (S) -2-hydroxyacid oxidase (EC 1. 1.3.15).
- the (D) -2-hydroxy oxidase of the present invention has a high activity on dalicholate, but also has an activity on D-form 2-hydroxy acids such as D-lactic acid, L-lactic acid has no activity.
- (D) -2-Hydroxyacid oxidase which selectively acts only on the D-form 2-hydroxy acid and does not act on the L-form 2-hydroxy acid, was first found in the present invention. In the case of the conventional glycolate oxidase, when the substrate is D-form, it hardly reacts or shows only low activity even when reacted, but the (D) -2-hydroxyacid oxidase of the present invention has been used for the first time. It became possible to efficiently react the D-form of 2-hydroxy acid. Therefore, the present invention
- (D) 2-hydroxy acid oxidase is intended not only to convert glycolic acid to dalioxylic acid, but also to react when the 2-hydroxy acid of D-form is a substrate or to selectively react with D-form. It is excellent in that it can also be used for optical division.
- the conventional glycolate oxidase (S) -2-hydroxyacid oxidase) is known to have a relatively high activity even for dalioxylic acid.
- the enzymes of the present invention show relatively low activity on dalioxylic acid. Therefore, the enzyme of the present invention is also advantageous in that conversion of glycolic acid to dalioxylic acid produces less by-products such as oxalic acid due to enzymatic oxidation of dalioxylic acid.
- the method for producing dalioxylic acid according to the present invention includes: a culture solution of the microorganism-derived (D) -2-hydroxyacid oxidase or a microorganism producing the enzyme; a microbial cell isolated from the culture solution;
- D microorganism-derived
- a microbial cell isolated from the culture solution One of the major features is that one of the treated products is allowed to act on dalicholate, and dalicholate is converted to dalioxylic acid and accumulated.
- the treated product of microbial cells is, for example, For example, it means a crude enzyme solution, freeze-dried cells, acetone-dried cells, or a crushed product of those cells.
- the term "crude enzyme solution” includes, for example, a physical method using glass beads, a biochemical method using enzymes, etc., after centrifuging cells from a culture solution and suspending them in an appropriate buffer. And a cell-free extract obtained by removing solids in the solution by centrifugation. Furthermore, an enzyme solution obtained by partially purifying the cell-free extract using a method commonly used by those skilled in the art, for example, dialysis, ammonium sulfate precipitation, or chromatography alone or in combination, is also referred to as a “crude enzyme solution”. " include. Furthermore, the processed microbial cells can be used by immobilizing the enzyme or the cells as they are by known means.
- the immobilization can be performed by a method known to those skilled in the art (for example, a crosslinking method, a physical adsorption method, an entrapment method, and the like).
- a microbial culture solution or cells can be used as they are without purifying the enzyme, which is an advantageous production method in industrial production.
- the reaction conditions vary depending on the enzyme used, the microorganism or its processed product, the substrate concentration, etc., but the reaction is carried out in the presence of oxygen, and the reaction temperature is 10 T: ⁇ 70 ° C.
- the reaction temperature is preferably from 0 ° C to 50, and the reaction pH is preferably from 4 to 12, and the pH is preferably from 6 to 10 from the viewpoint of optimum pH and pH stability.
- Example 1 Glycolic acid 10 g, yeast extract 0.1 g, ammonium nitrate 2 g, dipotassium hydrogen phosphate lg, sodium dihydrogen phosphate 1 g, magnesium sulfate heptahydrate 0.2 g, and calcium chloride 2 water 5 ml of an S medium (pH 7) having a composition of 0.1 g of the hydrate (all per liter) was placed in a test tube and sterilized with high-pressure steam. Then, 2 g of each soil sample collected from Japan was suspended in 10 ml of physiological saline and 0.2 ml of the supernatant was added to the S medium, and the suspension was added at 28 ° C for 3 to 7 days. Enrichment culture was performed.
- S medium pH 7
- the culture solution in which the bacteria had grown was applied in 0.1 ml portions to an S medium plate containing 2% agar, and cultured at 28 for 3 to 7 days. For the grown colonies, incubate each cell in a test tube with 5 ml of S medium at 28 ° C for 3 days with shaking, collect the cells by centrifugation, wash with saline, and add 10 OmM phosphate buffer. (PH 7)
- the suspension was suspended in 0.5 ml. 0.1 ml of the cell suspension 0.1 ml of 100 mM phosphate buffer containing 10 OmM glycolic acid, 1.34 mM 4-AA, 2.18 mM T ⁇ S, and 4 UZm 1 peroxidase And shaken at 28 ° C. for 2 hours.
- the one in which the reaction solution turned purple after shaking that is, the one in which hydrogen peroxide was generated by the reaction with dalicholate, was obtained as a glycolytic acid oxidizing activity-positive strain. '
- the inoculated strain of dalicholate oxidizing activity was inoculated into 5 ml of S medium, followed by shaking culture at 28 ° C for 2 days to obtain a precultured solution.
- 0.5 ml of the preculture solution was inoculated into 5 Oml of medium S medium, and cultured with shaking at 28 for 3 days.
- the cells were collected by centrifugation from 2 Oml of the obtained culture solution, washed with physiological saline, and suspended in 5 ml of a 100 mM phosphate buffer (pH 7) containing 100 mM dalicholate. It became cloudy and was shaken at 28 for 6 hours.
- KNK-GA1 strain isolated from soil (Deposited organization National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary, 1-chome, Tsukuba, Higashi, Ibaraki, Japan 1 Chuo No. 6 (zip code 305-8566), date of deposit On March 7, 2002, accession number FERM BP-8375) was placed in a 500-ml Sakaguchi flask and contained 5 g of glycolic acid, yeast extract lg, 3.5 g of dihydrogen phosphate, and hydrogen phosphate.
- the cells were collected from 10 ml of the obtained culture solution by centrifugation in a test tube, and the cells were suspended in 2 ml of 100 mM phosphate buffer (pH 7), and 1 M glycol was added to 0.9 ml of the cell suspension.
- An acid solution (0.1 lm 1) was added, and a shaking reaction was performed in a test tube at 28 ° C. for 16 hours.
- the obtained reaction solution was analyzed by high performance liquid chromatography. As a result, 3 OmM dalioxylic acid was produced.
- KNK-GA1 strain obtained by the method described in Example 2 (Deposited institution: National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary, Tsukuba East 1-chome, Ibaraki, Japan 1 Chuo No. 6 (Postal code 305- 8566), Deposit date: March 7, 2002, cells were collected by centrifugation from 50 ml of the culture solution of accession number FERM BP-8375), and washed with 0.1 M phosphate buffer (pH 7). The cells were suspended in 4 ml of 0.1 M phosphate buffer (pH 7). The cell suspension was crushed with a mini-bit beater (manufactured by BISPEC) and centrifuged to obtain a supernatant (cell-free extract).
- Example 6 0.1 ml of the cell-free extract after dialysis obtained in Example 4 containing 0.1 AA 1.34 mM, TOOS 2.19 mM and POD 6 U / m1 0.1 M phosphate buffer (pH 7) 0.05 ml was added, and 0.05 ml of a 10 OmM glycolic acid solution was further added. The mixture was shaken at 28 ° C for 2 minutes in a test tube. As a result, the reaction solution turned deep purple. 10 OmM glycol as control When 0.05 ml of 0.1 M phosphate buffer was added instead of the acid solution, the reaction solution did not discolor. From this, it was confirmed that hydrogen peroxide was generated by the oxidation of glycolic acid by the cell-free extract, and that the enzyme catalyzing the acidity of glycolic acid was oxidase. .
- Example 6 Example 6
- the cells were collected from the resulting culture by centrifugation and suspended in 1.2 L of 0.05 M phosphate buffer (PH7).
- the obtained cell suspension was disrupted by a dynomill, and then centrifuged to obtain 1.1 L of a supernatant.
- Add 62 ml of 5% protamine sulfate aqueous solution to the supernatant, and add After stirring for 0 minutes, the precipitate was removed by centrifugation to obtain 1.2 L of the supernatant.
- stirring 1.2 L of the obtained supernatant under ice-cooling with a stirrer gradually add ammonium sulfate, and when the saturation of the added ammonium sulfate is in the range of 25 to 40%.
- the sedimented protein was collected by centrifugation.
- ammonium sulfate precipitated protein was dissolved in 320 ml of 0.05 M phosphate buffer (PH7), dialyzed against 15 L of the same buffer, and then dialyzed with 0.05 M phosphate buffer (pH 7.2).
- the column was charged to a pre-equilibrated DEAE-Toyopearl 6 50M (Tosoichi) column (column diameter: 4 cm, height: 20 cm).
- the active fraction was eluted with m 1 Zhr and collected. After adding a 0.05 M phosphate buffer containing 1.2 M ammonium sulfate so that the concentration of ammonium sulfate becomes 0.6 M to the obtained active fraction, add 0.05 M ammonium sulfate.
- the dialyzed solution was pre-equilibrated with a 0.05 M phosphate buffer (pH 7.2) and a Super Q Toyopearl 650M (Tosoichi) column (column diameter: 2.4 cm, high was charged at 2 O cm) and eluted at a flow rate of 5 Oml / hr with a linear gradient of sodium chloride from 0 M to 0.3 M to collect the active fraction.
- the obtained active fraction was adjusted so that the concentration of ammonium sulfate became 0.4 M, and the first buffer was pre-equilibrated with a 0.05 M phosphate buffer (pH 7.2) containing 0.4 M ammonium sulfate.
- the concentration of ammonium sulfate in the active fraction was adjusted to 0.6 M, and then RESOURCE 15 PHE (Ame) pre-equilibrated with 0.05 M phosphate buffer (pH 7.2) containing 0.6 M ammonium sulfate rs ham pharmamaciabiotech) column (6 ml), and eluted at a flow rate of 2 ml / min with a 0.6 M-0 M ammonium sulfate linear concentration gradient to collect an active fraction.
- enzyme activity is measured in 10 OmM phosphate buffer or Tris buffer, 10 mM substrate such as glycolic acid, 4-AA 0.67 mM, T ⁇ OS 1.09 mM, POD 2 UZm 1, and enzyme
- the reaction was carried out by reacting 1.0 ml of the reaction solution containing 0.1 ml of the solution with 301: for 100 seconds and measuring the increase in absorbance at a wavelength of 555 nm.
- the enzyme of the present invention showed the substrate specificity shown in Table 2.
- the activity was measured using 1M phosphate buffer and 0.1M Tris-HCl buffer in a pH range of 5 to 10 using dalicholate as a substrate.
- Figure 3 shows the results. The optimum pH was 7-9.
- This purified enzyme was subjected to 10% SDS-polyacrylamide gel electrophoresis in the presence of 1% 2-mercaptoethanol, and its molecular weight was estimated from the relative mobility with the standard protein. As a result, the enzyme formed a single band at a position corresponding to a molecular weight of about 60,000 Da.
- the (D) -2-hydroxyacid oxidase of the present invention has the ability to efficiently convert glycolic acid to dalioxylic acid, and uses the enzyme or a microorganism that produces the enzyme to produce glycosolic acid. According to the method for producing glyoxylic acid from acid, it is possible to produce dalioxylic acid under mild conditions without producing by-products. Further, the (D) -2-hydroxyacid oxidase of the present invention is the first enzyme that selectively acts only on the D-form 2-hydroxyacid.
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Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP03730557A EP1507001A4 (en) | 2002-05-24 | 2003-05-22 | NOVEL (D) -2-HYDROXY ACID OXIDASE DERIVED FROM MICROORGANISM AND BIOCHEMICAL PROCESS FOR PRODUCING GLYCOXYLIC ACID |
AU2003242374A AU2003242374A1 (en) | 2002-05-24 | 2003-05-22 | Novel (d)-2-hydroxy acid oxidase derived from microorganism and biochemical process for producing glyoxylic acid therewith |
JP2004508284A JP4318637B2 (ja) | 2002-05-24 | 2003-05-22 | 微生物由来新規(d)−2−ヒドロキシ酸オキシダーゼ、およびそれを用いたグリオキシル酸の生化学的製造方法 |
US10/513,432 US20050170481A1 (en) | 2002-05-24 | 2003-05-22 | Novel (d)-2-hydroxy-acid oxidase derived from microorganism and biochemical process for producing glyoxylic acid therewith |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2002/150850 | 2002-05-24 | ||
JP2002150850 | 2002-05-24 |
Publications (1)
Publication Number | Publication Date |
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WO2003100043A1 true WO2003100043A1 (fr) | 2003-12-04 |
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Application Number | Title | Priority Date | Filing Date |
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PCT/JP2003/006367 WO2003100043A1 (fr) | 2002-05-24 | 2003-05-22 | Nouvelle (d)-2-hydroxy acide oxydase derivee d'un micro-organisme et procede biochimique permettant de produire de l'acide glycoxylique |
Country Status (5)
Country | Link |
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US (1) | US20050170481A1 (ja) |
EP (1) | EP1507001A4 (ja) |
JP (1) | JP4318637B2 (ja) |
AU (1) | AU2003242374A1 (ja) |
WO (1) | WO2003100043A1 (ja) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110452935A (zh) * | 2019-08-28 | 2019-11-15 | 精晶药业股份有限公司 | 一种酶法制备乙醛酸的方法 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1995001443A1 (en) * | 1993-07-01 | 1995-01-12 | E.I. Du Pont De Nemours And Company | Microbial transformants of hansenula that express glycolate oxidase and catalase |
US5834262A (en) * | 1992-01-06 | 1998-11-10 | E. I. Du Pont De Nemours And Company | Oxidation of glycolic acid to glyoxylic acid using a microbial cell transformant as catalyst |
-
2003
- 2003-05-22 EP EP03730557A patent/EP1507001A4/en not_active Withdrawn
- 2003-05-22 US US10/513,432 patent/US20050170481A1/en not_active Abandoned
- 2003-05-22 AU AU2003242374A patent/AU2003242374A1/en not_active Abandoned
- 2003-05-22 WO PCT/JP2003/006367 patent/WO2003100043A1/ja not_active Application Discontinuation
- 2003-05-22 JP JP2004508284A patent/JP4318637B2/ja not_active Expired - Fee Related
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5834262A (en) * | 1992-01-06 | 1998-11-10 | E. I. Du Pont De Nemours And Company | Oxidation of glycolic acid to glyoxylic acid using a microbial cell transformant as catalyst |
WO1995001443A1 (en) * | 1993-07-01 | 1995-01-12 | E.I. Du Pont De Nemours And Company | Microbial transformants of hansenula that express glycolate oxidase and catalase |
Non-Patent Citations (1)
Title |
---|
See also references of EP1507001A4 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN110452935A (zh) * | 2019-08-28 | 2019-11-15 | 精晶药业股份有限公司 | 一种酶法制备乙醛酸的方法 |
Also Published As
Publication number | Publication date |
---|---|
EP1507001A1 (en) | 2005-02-16 |
US20050170481A1 (en) | 2005-08-04 |
JPWO2003100043A1 (ja) | 2005-09-22 |
AU2003242374A1 (en) | 2003-12-12 |
JP4318637B2 (ja) | 2009-08-26 |
EP1507001A4 (en) | 2005-07-20 |
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