JPH0884592A - Method for biochemically producing glycol aldehyde - Google Patents

Abstract

PURPOSE: To biochemically obtain the compound useful as a raw material for α-amino acids, medicines, agricultural chemicals, etc., by treating ethylene glycol with an oxireductase capable of oxidizing the ethylene glycol to produce glycol aldehyde, and subsequently collecting the product. CONSTITUTION: The method for biochemically producing the glycol aldehyde comprises treating the ethylene glycol with an oxireductase having an ability to oxidize the ethylene glycol to produce glycol aldehyde, or a microorganism (e.g. Kluyveromyces marxianus IFO 0273) containing the oxireductase and belonging to the genus Pichia, Candida, Hansenula, Aspergillus, Penicillium, Neurospora, Pseudomonas, Acetobacter, Kluyveromyces, etc., and subsequently collecting the produced glycol aldehyde. The glycol aldehyde is useful as a raw material for α-amino acids, medicines, agricultural chemicals, photographic chemicals or polymers, or useful for fiber-treating agents, odorizing agents, deodorizing agents, etc.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to a biochemical method for producing glycolaldehyde. More specifically, by oxidizing ethylene glycol to ethylene glycol, and reacting a redox enzyme having the ability to produce glycolaldehyde or a microorganism containing the redox enzyme,
The present invention relates to a biochemical production method of glycol aldehyde, which comprises converting ethylene glycol to glycol aldehyde and collecting the product.

Glycolaldehyde is an α-amino acid,
It is a compound useful as a raw material for medicines, agricultural chemicals, photographic agents, special polymers, and as a fiber treatment agent, odorant, and deodorant.

[0003]

2. Description of the Related Art Conventionally, glycol aldehyde is a method in which ethylene glycol and water vapor are passed through a copper or copper / zinc catalyst (JP-A-61-69740), silver, copper, silver oxide, copper oxide, etc. and silicon. , A compound using phosphorus or a boron compound is used (JP-A-62-45548), and ethylene glycol is introduced in a gas layer into a copper-zinc composite catalyst layer filled in a reaction tube. However, it has been industrially manufactured by a chemical synthesis method such as a method of condensing the produced gas by a cooler connected to the reaction tube (JP-A-3-279342). Was not manufactured until.

[0004]

However, when glycol aldehyde is produced by a chemical synthesis method,
Since heavy metals and reactions at high temperatures are required, and a large amount of acid is by-produced, it is necessary to remove and purify them, which requires complicated operations. Therefore, there has been a demand for a method for producing glycolaldehyde that produces less by-products under mild conditions.

[0005]

Under these circumstances, the present inventors have found that the method does not require complicated operations and that the production of by-products is extremely small under mild conditions, that is, biochemical methods. An attempt was made to produce glycolaldehyde by the method.

[0006] So far, as biochemical studies on the oxidation of alcohols, the existence of oxidoreductases that act on certain monovalent or trivalent alcohols has been known [Y.
Tani, T. Miya and K. Ogata, Agric. Biol. Chem., 36
Volume, 76-83 (1972), H. Sahm and F. Wagner, Eur.
J. Biochem., 36, 250-256 (1973), N. Kato, Y.
Omori, Y. tani and K. Ogata, Eur. J. Biochem., 64
Volume, 341-350 (1976), CR Couder and J. Baratt
i, Agric. Biol. Chem., 44, 2279-2289 (1980),
T. Uwajima, H. Akita, K. Ito, A. Mihara, K. Aisak
a and O. Terada, Agric. Biol. Chem., Volume 43, 2633 ~ 2
634 (1979)].

Regarding the oxidoreductase acting on the dihydric alcohol, only the possibility of its existence is suggested [A. Boronat, E. Caballero and J. Aguilar, J. Bact.
eriol., 153, 134-139 (1983)], the identification of the reaction product was not performed at all.

The present inventors have been engaged in research on oxidoreductases produced by microorganisms for many years, and as a result of earnestly examining the reaction of oxidoreductases produced by microorganisms with ethylene glycol, glycol aldehyde and glyoxal, In a redox enzyme or a microorganism containing the redox enzyme having the ability to oxidize ethylene glycol and produce glycolaldehyde, it is newly found that glycolaldehyde can be accumulated from ethylene glycol, and using these enzymes or microorganisms, We have developed a method for producing and accumulating glycolaldehyde from ethylene glycol.

The present invention has been completed based on these findings. That is, ethylene glycol is converted to glycol aldehyde by reacting with oxidoreductase having the ability to oxidize ethylene glycol to ethylene glycol and produce glycol aldehyde, or a microorganism containing the oxidoreductase, and collecting this. Is a biochemical production method of glycol aldehyde.

The present invention makes it possible for the first time to efficiently produce glycolaldehyde under mild conditions.

The chemical reaction formulas for producing glycolaldehyde from the ethylene glycol of the present invention by the oxidoreductase reaction are shown in Formulas 1 and 2.

[0012]

(Equation 1)

[0013]

[Formula 2]

The reaction represented by the above formula 1 is a case where a dehydrogenase or a microorganism containing the dehydrogenase is mediated, and the above formula 2 is a case where an oxidase or a microorganism containing the oxidase is mediated. Recognized in.

Hydrogen peroxide is produced in the enzyme reaction of the above formula 2, and this hydrogen peroxide destabilizes the oxidase, oxidizes the produced glycol aldehyde, etc., so that the amount of glycol aldehyde produced is increased. May decrease. However, even in this case, hydrogen peroxide can be removed by adding catalase during the reaction, and the amount of accumulated glycolaldehyde can be increased. In addition, an enzyme that has the ability to oxidize ethylene glycol to produce glycol aldehyde may further oxidize glycol aldehyde to glyoxal. However, by selecting appropriate conditions for accumulating glycol aldehyde, glyoxal of glycol aldehyde can be selected. It is also possible to suppress the oxidization to glycerol and increase the amount of accumulated glycolaldehyde.

The ethylene glycol, which is the starting substance of the present invention, is a typical dihydric alcohol, and industrially, ethylene chloride is reacted with hydrochloric acid water to obtain ethylene chlorohydrin, which is hydrogen carbonate. Hydrolyzed in an autoclave with sodium or sodium carbonate solution, or
It is produced on the market by heating ethylene chlorohydrin with alkali hydroxide to give ethylene oxide, which is hydrolyzed by reacting it with dilute sulfuric acid.

In screening the enzymes and microorganisms that can be used in the above reaction, the present inventors first established a method for quantifying the reaction products glycolaldehyde and glyoxal, and further, in the above reaction,
A method for quantifying glyoxylic acid was also established to confirm that glyoxylic acid (OHCCOOH) was not produced.

Experimental Example 1 The amount of glycol aldehyde was determined by [MA Paz, OOB
lumenfeld, M. Rojikind, E. Henson, C. Furfin and
P. Gallop, Arch. Biochem. Biophys., 109, 547-55
9 (1965)], etc., and the method was performed under the following conditions. 0.
For 10 μl of 5 mM to 2.5 mM glycolaldehyde solution
0.1 M glycine hydrochloric acid buffer (pH 4.0) 150 μl, 1% MBTH
[3-methyl-2-benzothiazolinone / hydrazone /
Hydrochloride (3-methyl-2-benzothiazolinone hyd
razone hydrochloride)] solution (60 μl), 0.2% ferric chloride solution (750 μl) is added, and glycolaldehyde derivative-
2 was produced, and the relationship between the absorption spectrum and the absorbance at 610 nm and the concentration of glycolaldehyde was examined. As a result, the absorbance curve as shown in FIG. 1 and the relationship between the absorbance and glycolaldehyde concentration as shown in FIG. 2 were obtained, and it was shown that glycolaldehyde can be quantified under these conditions.

Experimental Example 2 Glyoxal was quantified by [MA Paz, OO Blumenf
eld, M. Rojikind, E. Henson, C. Furfin and P. Gallo
p, Arch. Biochem. Biophys., 109, 547-559 (196
5)] and other methods were improved and carried out under the following conditions. 0.5 mM ~
To 10 μl of 2.5 mM glyoxal solution, 750 μl of 0.1 M glycine-HCl buffer (pH 4.0) and 300 μl of 1% MBTH solution were added to generate glyoxal derivative-1, and its absorption spectrum and absorbance at 380 nm and glyoxal concentration were measured. Examined the relationship. As a result, the relationship between the absorbance curve as shown in FIG. 1 and the absorbance and glyoxal concentration as shown in FIG. 2 was obtained, and it was shown that glyoxal can be quantified under these conditions.

Experimental Example 3 Quantification of glyoxylic acid was carried out by [MA Paz, OO Blumenf
eld, M. Rojikind, E. Henson, C. Furfin and P. Gallo
p, Arch. Biochem. Biophys., 109, 547-559 (196
5)] and other methods were improved and the same conditions as in Experimental Examples 1 and 2 were used. As a result, the absorbance curve shown in FIG. 1 and the relationship between the absorbance and the glyoxylic acid concentration shown in FIG. 2 were obtained, and it was shown that glyoxylic acid can be quantified under these conditions.

Experimental Example 4 In Experimental Examples 1 to 3, glyoxal was colored yellow when 1% MBTH solution was added, but was not colored blue even when 0.2% ferric chloride solution was added. Glycolic aldehyde and glyoxylic acid were colorless when the 1% MBTH solution was added, and became blue by adding the 0.2% ferric chloride solution. Therefore, it was possible to separately measure glyoxal from glycolaldehyde and glyoxylic acid, but the absorption curves of the glycolaldehyde and glyoxylic acid derivatives were similar and it was difficult to distinguish them. Then, the derivatives of these three kinds of compounds were prepared according to Experimental Examples 1 to 3, and the reverse phase using a C ₁₈ column was used.
Fractionation of the three compounds was performed by HPLC chromatography.
Regarding glyoxylic acid, solutions of derivative-1 and derivative-2 before and after addition of 0.2% ferric chloride solution were subjected to HPLC. As a result, as shown in FIG. 3, without addition of ferric chloride, glyoxal (derivative-1) had peaks at 34.0 minutes and 34.6 minutes, respectively.
In 1), a peak was recognized around 12.1 minutes. In the derivative of ferric chloride added liquid, glycolaldehyde (derivative-2) was around 16.4 minutes, glyoxylic acid (derivative-2).
However, a peak was obtained at around 22.8 minutes, and it was possible to separately quantify three kinds of compounds.

Since a method for quantifying the three kinds of compounds could be established in this way, oxidoreductase having the ability to oxidize ethylene glycol and produce and accumulate glycolaldehyde was screened. As a result, it was revealed that the oxidase has the ability to oxidize ethylene glycol into alcohol oxidase and glycerol oxidase to produce and accumulate glycol aldehyde. Alcohol oxidases include Pichia sp.
pastoris) -derived alcohol oxidase, Candida s
p. No. SS104 (The mycological properties of this bacterium are described in Koshien University Bulletin,
No. 18, (A), 23-29 (1990), and has been deposited as FERM-P No. 13702 at the Institute of Biotechnology, Institute of Biotechnology, AIST. ) According to the literature, alcohol oxidase derived from the genus Candida, alcohol oxidase derived from the genus Hansenula, which is a product of Sigma, and the like are exemplified.On the other hand, as glycerol oxidase, literature and Japanese Examined Patent Publication No. Aspergillus japonicus KY-45 ATCC N described in Kosho 60-49477
o. 1042 derived glycerol oxidase from Aspergillus, and further Penicillium sp. KY868 NR
Examples include penicillium-derived glycerol oxidase derived from RL 11339.

Next, the present inventor extensively searched for microorganisms containing an oxidoreductase which has the ability to oxidize ethylene glycol and produce glycolaldehyde. As a result, it was found that at least the microorganisms exemplified below have the ability. That is, Pichia (for example, Pichia p
The astoris JCM 3650 is exemplified. ), Candida spp (for example, Candida sp. No. SS104 FERM P-13702 is exemplified), and Hansenula spp (for example, Hansenula polymorp).
ha ATCC 26012 is exemplified. ), Aspergillus (for example, Aspergillus japonicus KY-45 ATCC 1042 is exemplified.), Penicillium (for example, Penicill)
ium sp. KY868 NRRL 11339 is exemplified. ), Neurospora (for example, Neurospora crassa IFO 6068 is exemplified), Pseudomonas (for example, Pseudomona).
s sp. GOX-201 FERM P-14328 is exemplified. ), Acetobacter (for example, Acetobacter pasteurianus IFO)
3222 is exemplified. ) And Kluyveromyces spp. (For example, Kluyveromyces marxianus IFO 0273 is exemplified).

Further, the present inventors conducted the following experiment to examine the relationship between the concentration of ethylene glycol and the production of glycolaldehyde and glyoxal using oxidoreductase.

Experimental Example 5 To 9 ml of 0.2 M TES buffer (pH 7.0) containing 1.1 M to 6.6 M ethylene glycol, 60 u of glycerol oxidase derived from Aspergillus and 130 ku of catalase were added (final reaction solution volume: 10 ml). ), And reacted at 20 ° C. for 24 hours. MBTH derivative was prepared from this reaction solution according to Experimental Examples 1 and 2,
The results of determining the concentrations of glycolaldehyde and glyoxal are shown in Table 1 and FIG.

[0026]

[Table 1]

Experimental Example 6 As in Experimental Example 5, the ethylene glycol concentration during the reaction was 2
To a reaction solution prepared so as to have a concentration of M to 10 M, 1000 u of Pichia-derived alcohol oxidase and 130 ku of catalase were added and reacted at 20 ° C for 4 hours. The results are shown in Table 2 and FIG.

[0028]

[Table 2]

As shown in Table 1 and FIG. 4 or Table 2 and FIG. 5 of Experimental Examples 5 and 6, the action of oxidoreductase having the ability to oxidize ethylene glycol to the substrate ethylene glycol to produce glycol aldehyde. It was also revealed that it is possible to generate glycolaldehyde, and further increasing the concentration of ethylene glycol can reduce the production ratio of glyoxal to glycolaldehyde.

Experimental Example 7 Various buffer solutions containing 2.0 M ethylene glycol, that is,
TES 〔N-Tris (hydroxymethyl) methyl-2-aminoethane s
ulphonic acid] buffer, Tris-HCl buffer, BES [N, N-Bis (2-hydroxymethyl) -2-aminoethane su
lfonic acid] buffer, MOPS (3-Morpholinopropane sul
fonic acid) buffer solution, AMPD 〔2-Amino-2-methyl-1,3prop
andiol] buffer solution, boric acid (Na ₂ B ₄ O ₇ · HCl) buffer solution, and phosphate buffer solution (0.9 ml) were added with 50 μl of Pichia-derived alcohol oxidase and 13 ku of catalase (final reaction solution volume). : 1 ml) and reacted at 20 ° C. for 4 hours. An MBTH derivative was prepared from this reaction solution according to Experimental Examples 1 and 2, and the results of determining the concentrations of glycolaldehyde and glyoxal are shown in Table 3.

[0031]

[Table 3]

As is clear from Table 3, in any of the buffer solutions, a large amount of glycol aldehyde could be produced and accumulated from ethylene glycol.

Further, the present inventor conducted Experimental Examples 8 to 11 in order to examine the influence on the glycolaldehyde formation by changing the pH during the reaction.

Experimental Example 8 0.2 M Tris-HCl buffer solution (pH 7.
0-9.0) to 2.0 M ethylene glycol, and add 25 μl of Pichia-derived alcohol oxidase and 2.6 ku of catalase (final reaction volume: 1 ml) at 20 ° C.
And reacted for 4 hours. The MBTH derivative was prepared from this reaction solution according to Experimental Examples 1 and 2, and the results of determining the concentrations of glycolaldehyde and glyoxal are shown in Table 4.

[0035]

[Table 4]

Experimental Example 9 Dissolved in 0.2 M 2 amino-2-methyl-1,3 propanediol (AMPD) buffer solution (pH 7.5-9.5) at various pHs to give 2.0 M ethylene glycol, and derived from Pichia Alcohol oxidase (25 u) and catalase (2.6 ku) were added (final reaction solution volume: 1 ml) and reacted at 20 ° C for 4 hours. The MBTH derivative was prepared from this reaction solution according to Experimental Examples 1 and 2, and the results of determining the concentrations of glycolaldehyde and glyoxal are shown in Table 5.

[0037]

[Table 5]

Experimental Example 10 0.2 M Tris-HCl buffer solution (pH 7.
0-9.0) to 2.0 M ethylene glycol, and add 32 u of Aspergillus-derived glycerol oxidase and 2.6 ku of catalase (final reaction volume: 1
ml) and reacted at 20 ° C. for 4 hours. Experimental Example 1 from this reaction solution
Table 6 shows the results of preparing the MBTH derivatives according to 2 and 2 and determining the concentrations of glycolaldehyde and glyoxal.

[0039]

[Table 6]

Experimental Example 11 Dissolved in 0.2 M 2 amino-2-methyl-1,3 propanediol (AMPD) buffer solution (pH 7.5-9.5) at various pHs to give 2.0 M ethylene glycol, and derived from Aspergillus Glycerol oxidase 32 u, catalase 2.6
ku was added (final reaction solution volume: 1 ml), and the mixture was reacted at 20 ° C. for 4 hours. The MBTH derivative was prepared from this reaction solution in accordance with Experimental Examples 1 and 2, and the results of determining the concentrations of glycolaldehyde and glyoxal are shown in Table 7.

[0041]

[Table 7]

As is clear from Tables 4 to 7, pH 7-9.
5 That is, by reacting an enzyme in the neutral to alkaline range, a large amount of glycolaldehyde can be produced and accumulated from ethylene glycol, and by using a buffer solution in the alkaline range, the reaction product glycolaldehyde can be It became clear that the production ratio of glyoxal to

The conditions under which the enzyme acts in the biochemical method for producing glycolaldehyde of the present invention will be described. First, the reaction time is not particularly limited as long as the condition under which the enzyme acts, but is preferably 30 minutes to 30 days. Is. The reaction temperature is also not particularly limited, but the preferable reaction temperature is 0 to 40 ° C. The pH condition during the reaction is not particularly limited as long as the enzyme can act on the pH. Usually, the pH is 3 to 10
And preferably pH 7-10.

The present invention will be described in more detail with reference to the following examples, but the present invention is not limited to these examples. The method for measuring the activity of various enzymes in the examples was the method described in the above document for alcohol oxidase and the method described in the above document for glycerol oxidase. The catalase was performed according to JP-A-2-76579.

[0045]

【Example】

Example 1 Acetobactor past
eurianus) IFO 3222 5% ethylene glycol, 2% coast liquor (CSL), 0.2% ammonium sulfate, 0.1% dipotassium phosphate, 0.1% disodium phosphate, 0.02% magnesium sulfate, 0.01% calcium chloride, 0.1% As a result of culturing in a yeast extract (pH5.5) medium at 30 ° C for 5 days, the concentrations of glycolaldehyde and glyoxal produced in the culture solution were 130 mM and 2 mM, respectively, and the production ratio of glyoxal was glycolaldehyde. Was 1.5%.

Example 2 Kluyveromyces ma
rxianus) IFO 0273 2% 1,2-propanediol, 0.5
% CSL, 0.2% ammonium sulfate, 0.1% dipotassium phosphate, 0.1%
Disodium phosphate, 0.02% magnesium sulfate, 0.01%
After culturing the cells in a medium (100 ml) consisting of calcium chloride and 0.1% yeast extract (pH 7.0) for 2 days at 30 ° C and washing the cells, 20 ml of 0.2 M TES buffer solution containing 1 M ethylene glycol ( It was suspended in pH 7.0) and reacted at 20 ° C for 15 hours. As a result, the production of 101 mM glycolaldehyde was confirmed, but the production of glyoxal was not observed at all.

Example 3 0.2 M TES buffer containing 6.6 M ethylene glycol (pH
7.0) To 9 ml, 60 u of Aspergillus-derived glycerol oxidase and 130 ku of catalase were added (final reaction solution volume: 10 ml), and reacted at 20 ° C for 24 hours. MBTH derivatives were prepared from this reaction solution according to Experimental Examples 1 and 2, and the concentrations of glycolaldehyde and glyoxal were determined.
The amount of glycolaldehyde and glyoxal produced is 0.
238 M and 0.005 M, and the amount of glyoxal produced was about 2.1% of glycol aldehyde, and this glycol aldehyde was collected by a conventional method.

Example 4 0.2 M TES buffer containing 6.6 M ethylene glycol (pH 7.
0) To 9 ml, add 75 u or 300 u of Aspergillus-derived glycerol oxidase and 130 ku of catalase,
(Final reaction liquid volume: 10 ml) and reacted at 20 ° C. for 10 hours. As a result, as shown in Table 8, the amount of glyoxal produced was 5% or less of glycol aldehyde in each case.

[0049]

[Table 8]

Example 5 Under the same conditions as in Example 4, 675 u of glycerol oxidase derived from Aspergillus was reacted at 20 ° C. for 48 hours. As a result, the amounts of glycolaldehyde and glyoxal produced were 0.645 M and 0.030 M, the amount of glyoxal produced was about 4.6% of glycolaldehyde, and glycolaldehyde accumulated due to the oxidation of ethylene glycol even in the long-term reaction.

Example 6 As in Example 3, the ethylene glycol concentration during the reaction was 6
The mixture was adjusted to M, 1000 μl of Pichia-derived alcohol oxidase and 130 ku of catalase were added, and reacted at 20 ° C for 4 hours. As a result, the amount of glycolaldehyde and glyoxal produced was 0.350 M and 0.0097 M, and the amount of glyoxal produced was about 2.8% that of glycolaldehyde.
Therefore, a significant amount of glycol aldehyde was accumulated by the oxidation of ethylene glycol.

Example 7 1M Tris-HCl buffer containing 1M ethylene glycol (p
H9.0) Alcohol oxidase derived from Pichia
Add 000 u and 130 ku of catalase (final reaction volume: 1
(0 ml), the pH was kept around 9 and reacted at 5 ° C. for 30 hours. As a result, 0.756 M glycolaldehyde and 0.028 M
Generation of glyoxal was confirmed, and when it was allowed to act in an alkaline region, the amount of glyoxal produced was 4% or less of glycol aldehyde even when 75% or more of ethylene glycol was oxidized.

Example 8 1M Tris-HCl buffer containing 1M ethylene glycol (p
H9.0) glycerol oxidase 320 u from Aspergillus genus 320 u and catalase 130 ku were added to 9 ml (final reaction volume: 10 ml), and the pH was kept around 9 at 48 ° C at 5 ° C.
Reacted for hours. As a result, the amounts of glycolaldehyde and glyoxal produced were 0.832 M and 0.013 M, and when they were allowed to act in an alkaline region, the amount of glyoxal produced was about 2% or less of glycolaldehyde even if 80% or more of ethylene glycol was oxidized. Met.

Example 9 223 u of alcohol oxidase derived from Pichia was fixed on CNBr-Sepharose (wet weight 1 g) by a conventional method and packed in a column. 5 M containing 130 ku of catalase (Boehringer Mannheim product, from beef liver) in this column
20 ml ethylene glycol solution (0.2 M TES buffer (pH 7.
0)] was circulated and reacted for 4 days. As a result, 165 mM glycol aldehyde was produced and no glyoxal was produced.

Example 10 1060 u of alcohol oxidase derived from Pichia was fixed on CNBr-Sepharose (dry weight: 1.15 g) by a conventional method, and 4
Loaded into a ml column. 50 ml of a 2 M ethylene glycol solution [0.5 M Tris-HCl buffer (pH 9.0)] containing 1,300 ku of catalase (product of Boehringer Mannheim, derived from beef liver) was circulated in this column and reacted at 5 ° C for 20 days. As a result, 570 mM glycolaldehyde was produced, and the production concentration of glyoxal was 13.8 mM.

[0056]

INDUSTRIAL APPLICABILITY The present invention is to biochemically produce glycolaldehyde by using an oxidoreductase having the ability to oxidize ethylene glycol to produce glycolaldehyde or a microorganism containing the oxidoreductase. This has the advantage that glycolaldehyde can be produced under mild conditions and with a small amount of by-product formation.

[Brief description of drawings]

FIG. 1 shows absorbance curves of each derivative-1 (shown by a solid line) and derivative-2 (shown by a broken line) of glycolaldehyde, glyoxal and glyoxylic acid. In the figure, numeral 1 indicates water, 2 indicates 15 mM glyoxal, 3 indicates glyoxylic acid, and 4 indicates glycol aldehyde.

FIG. 2 Glyoxylic acid, glyoxal derivative-1
(Measured at a wavelength of 380 nm and indicated by a solid line) and respective derivative curves of glycolaldehyde and glyoxylic acid-2 (measured at a wavelength of 610 nm and indicated by a broken line) are shown. In the figure, white circles represent glyoxylic acid, black circles represent glyoxal, and white squares represent glycolaldehyde.

FIG. 3 is a reverse-phase liquid chromatograph of various derivatives. Peaks 1 and 2 in the figure show each derivative-1 of glyoxylic acid and glyoxal-1 (measured at a wavelength of 380 nm), and peaks 3 and 4 show each induction-2 of glycolaldehyde and glyoxylic acid (the wavelength is 610 nm). (measured in nm)
Are shown respectively.

FIG. 4 shows the relationship between the production ratio of glycolaldehyde to glyoxal and the concentration of substrate ethylene glycol in Experimental Example 5.

FIG. 5 shows the relationship between the production ratio of glycolaldehyde to glyoxal and the concentration of substrate ethylene glycol in Experimental Example 6.

Continuation of the front page (51) Int.Cl. ⁶ Identification number Office reference number FI Technical display location (C12P 7/26 C12R 1:78) (C12P 7/26 C12R 1:66) (C12P 7/26 C12R 1: 80) (C12P 7/26 C12R 1:38) (C12P 7/26 C12R 1:02)

Claims (5)

[Claims] 1. An ethylene glycol is converted into glycol aldehyde by reacting it with a redox enzyme having the ability to oxidize ethylene glycol into ethylene glycol and producing glycol aldehyde, or a microorganism containing the redox enzyme. A biochemical method for producing glycolaldehyde, which comprises: 2. The biochemical method for producing glycolaldehyde according to claim 1, wherein the oxidoreductase having the ability to oxidize ethylene glycol to produce glycolaldehyde is a dehydrogenase. 3. The biochemical method for producing glycolaldehyde according to claim 1, wherein the oxidoreductase having an ability to oxidize ethylene glycol to produce glycolaldehyde is an oxidase. 4. The claim wherein the microorganism is a strain belonging to any one of Pichia, Candida, Hansenula, Aspergillus, Penicillium, Neurospora, Pseudomonas, Acetobacter, and Kluyveromyces. Item 10. A biochemical method for producing glycolaldehyde according to Item 1. 5. The biochemical method for producing glycolaldehyde according to claim 3, wherein the oxidase having the ability to oxidize ethylene glycol to produce glycolaldehyde is alcohol oxidase or glycerol oxidase.