JP2679145B2 - Oxyketone manufacturing method - Google Patents

Description

TECHNICAL FIELD The present invention relates to a method for producing an oxyketone, and more particularly to a method for producing a corresponding oxyketone from butanediol or pentanediol by using a specific microorganism.

4-Hydroxy-2-butanone, which is a kind of oxyketone, is useful as a raw material for various organic synthesis including methyl vinyl ketone.

(Problems to be Solved by the Prior Art and Invention) As a method for producing 4-hydroxy-2-butanone, which is a type of oxyketone, a method of reacting acetone and formaldehyde is generally used. Since hydroxy-2-butanone is highly reactive with the raw material formaldehyde, various by-products are formed and the yield is low.

With respect to a method for producing a corresponding oxyketone from butanediol or pentanediol using a microorganism, a method using Corynebacterium or Saccharomycopsis has been proposed (Japanese Patent Laid-Open No. 63-49089). The inventors completed a present invention by conducting a search for a wide range of microorganism groups in order to obtain microorganisms having further excellent ability.

(Means for Solving the Problems) As described above, the present inventors have developed a method for producing a corresponding oxyketone using a microorganism from butanediol or pentanediol that is industrially inexpensively available as a raw material. Various studies were conducted to make it.

As a result, Acinetobacter, Achromobacter, Aeromonas, Agrobacterium, Alcaligenes, Arthrobacter, Aureobacterium, Bacillus, Brevibacterium, Citrobacter, Enterobacter, Escherichia Genus, Flavobacterium, Klebzilla, Lactobacillus, Methylobacterium, Micrococcus, Mycobacterium, Paracoccus, Pimelobacter, Proteus, Serratia, Vibrio, Xanthomonas, Ambrosiojima , Arthrosuscus, Botilioascus, Brettanomyces, Candida, Citelomyces, Clavispora, Cryptococcus, Debaryomyces, Deckera, Dipoduscus, Endomyces, Fi Genus Basidium, genus Geotricum, genus Guiliermondera, genus Hansenula, genus Hansenia spora, genus Issachianchia, genus Kluyveromyces, genus Rodellomyces, lipomyces, genus Nicotiana, patissolen, Pichia, saccharomyces, saccharomyces , Schizoblast sporion, Schizosaccharomyces, Siwaniomyces, Selenojima, Serenotila, Stefanoascus, Sterigmatomyces, Polydiobolus, Silingospora, Torulaspora, Torlopsis, Trichosporon The inventors have found that microorganisms belonging to the genera Trigonopsis, Wickelamia, Wickelamiella, Wingae, or Tigosaccharomyces can achieve the object of the present invention, and completed the present invention. That is, the present invention provides at least one diol selected from butanediol and pentanediol as Acinetobacter, Achromobacter, Aeromonas, Agrobacterium, Alcaligenes, Arthrobacter, Aureobacteria. Genus, Bacillus, Brevibacterium,
Citrobacter, Enterobacter, Escherichia, Flavobacterium, Klebzilla, Lactobacillus, Methylobacterium, Micrococcus,
Mycobacterium, Paracoccus, Pimelobacter, Proteus, Serratia, Vibrio, Xanthomonas, Ambrosiojima, Arthrosus,
Botilioascus genus, Brettanomyces genus, Candida genus, Citalomyces genus, Clavispora genus, Cryptococcus genus, Debaryomyces genus, Deckera genus, Dipodascus genus, Endomyces genus, Philobasidium genus,
Geotricum genus, Guilliermondera genus, Hansenula genus, Hansenia spora genus, Issachenchia genus, Kluyveromyces genus, Roderomyces genus, Lipomyces genus,
Methinicovia, Patissolen, Pichia, Saccharomyces, Saccharomyces, Schizoblast sporion, Schizosaccharomyces, Shiwaniomyces, Serenodima, Serenotila, Stefanoascus, Sterigmatomyces , Polydiobolus,
Silingospora, Torluspora, Torlopsis,
To provide a method for producing an oxyketone, which is characterized by trichosporone, Trigonopsis, Wickelamia, Wickelamiella, Wingae, or Tigosaccharomyces, which is characterized by being oxidized using a microorganism capable of oxidizing the diol. .

The butanediol used as a raw material in the present invention contains 1,3-
In addition to butanediol, there are 1,2-butanediol, 2,3-butanediol, and the like. Pentanediol includes 1,2-pentanediol, 1,3-pentanediol, 1,4-pentanediol, and 2,3. -Pentanediol and the like.

The microorganisms that can be used in the present invention include Acinetobacter, Achromobacter, Aeromonas, Agrobacterium, Alcaligenes, Arthrobacter, Aureobacteria, Bacillus, Brevibacterium, Citrobacter, Enterobacter, Escherichia, Flavobacterium, Klebzilla, Lactobacillus, Methylobacterium, Micrococcus, Mycobacterium, Paracoccus, Pimelobacter, Proteus, Serratia, Vibrio genus, Xanthomonas genus, Ambrosiojima genus, Arthroascus genus, Botilioascus genus, Bretanomyces genus, Candida genus, Citellomyces genus, Clavispora genus, Cryptococcus genus, Debaryomyces genus, Deckera, Dipodus Scan the genus, Endomycetales, Philo Basi di Umm genus, Geotorikamu genus, Gui Riel Mont de la spp., Hansenula, Hanseniasupora genus, Issa Chen Kia genus Kluyveromyces, Roderomaisesu genus,
Lipomyces spp, Methinicovia spp, Pachisolen spp, Pichia spp, Saccharomyces spp, Saccharomyces spp, Schizoblast sporion spp, Schizosaccharomyces spp, Swaniomyces spp, Selenojima spp, Serenotila spp, Stefanoascus spp, Sterigmat Myces, Spolidiobolus, Siringospora, Torulaspora,
Tolulopsis, Trichosporon, Trigonopsis, Wickelamia, Wickelamiella, Wingae, or Tigosaccharomyces, which has the ability to oxidize butanediol or pentanediol to form the corresponding oxyketone, is used. However, as a specific example, Acinetobacter calcoaceticus IFO1
255, Achromobacter pestifer
pestifer) IFO ATCC 23584, Aeromonas hydrophila subspecies hydrofila (Aeromona
s hydrophila subsp.hydrophila) IFO 3820, Agrobacterium radiobactor
acter) IFO 12664, Alcaligenes Species (A
lkaligenes sp.) IAM 1015, Arthrobacter citreus IAM 12341, Aureobacterium testaceum
um) IFO 12675, Bacillus subtilis
tilis) IFO 13169, Brevibacterium linens (B
revibacterium linens) IFO 12141, Citrobacter
Freundy (citrobacter freundii) AHU 1534,
Enterobacter cloaca
e) IFO 3320, Escherichia col
i) IFO 3301, Flavobacterium lutescens (Fl
avobacterium lutescens) IFO 12997, Krebsira
Klebsiella pneumoniae IFO 3319,
Lactobacillus planta
rum) IFO 3070, Methylobacterium extorquens IAM 1081, Micrococcus lutesus IFO 30
67 、 Mycobacterium smegmatis
smegmatis) IFO 3153, Paracoccus denitrificans IFO 12442, Pimelobacter simplex I
FO 12069, Proteus mirabili
s) IFO 3849, Serratia Marcessens
arcesens) IFO 304, Vibrio anguiralum (Vibr
io anguillarum) IFO 12710, Xanthomonas campestr
is pv. oryzae) IAM 1657, Ambrosiozyma ciatricosa IFO 1846, Arthroascus javanensi
s) IFO 1848, Botryoascus synnaedendrus IFO 1604, Brettanomyces bruxellens
is) IFO 0628, Candida uti
lis) IFO 0626, Citeromyces matsuritensis (Ci
teromyces matritensis) IFO 0954, Clavispora lusitaniae IFO 1019, Cryptococcus albidus D
SM7019, Debaryomyces
hansenii) IFO 0083, Deckera Burgselensis (D
ekkera bruxellensis) IFO 1590, dipodascus
Alpodus (Dipodascus albidus) IFO 1984, Endomyces decipiensu IFO
0102, Filobasidium capsulighenum
idium capsuligenum) IFO 1119, Geotrichum candidum IFO 4601, Guilliermondella sel
ennospora) IFO 1850, Hans Anura (Hans)
enula anomela) IFO 0707, Hansenia guilliermondii I
FO 0683, Issatchen Kia Scutrate (Issatchen
kia scutulate) IFO 10069, Kluyveromyces lactis IFO 1267, Roderomyces elongisporu
s) IFO 1676, Lipomyces Starkey (Lypomyces)
starkeyi) IFO 1289, Metinicovia Pulcherima (M
etschnikowia pulcherrima) DSM 79336, pachisolene
Pachsolen tannophilus IFO 1007,
Pichia heedii IFO 10020, Saccharomyces serevisi
ae) IFO 0718, Saccharomycodes ludwigii IFO 0798, Schizoblastosporion koba
yashii) IFO 1644, Schizosaccharomyces pombe (S
chizosaccharomyces ponbe) IFO 0363, Schwanniomyces ludwigii IFO
1841, Selenozyma peltate
IFO 185, Selenotila pel
tate) DSM 70579, Stephanoascus ciferrii IFO 1854, Sterigmatomyces elviae DSM
70852, Sporidiobora salmonicolor (Sporid
iobolus salmonicolor) IFO 1845, Siringospora
Albicans (Syringospora albicans) IFO 1856, Torulaspora delbruec
kii) IFO 1083, Torrlopsis nitratophilia (To
rulopusis nitratophilia) DSM 70649, Trichosporon cutaneum IFO 1198,
Trigonopusis variabi
lis) IFO 0755, Wickerhamia fluoresces IFO 1116, Wickermiella domercquii IFO
1857, Wingea robertsii
IFO1857 or Chigosaccharomyces bisporus (Zygo
saccharomyces bisporus) DSM 70415 and the like. These mutants can also be used.

Among the above, microorganisms with ATCC number are American
Catalog o published by Type Culture collection (ATCC)
f Bacteria Phages rDNA Vectors 16th Edition (1985) and is available from the ATCC.

The microorganisms with the IFO number are the fermentation research institute (IF).
O), List of Cultures, 8th Edition, Volume 1 (1988), and can be obtained from the IFO.

Microorganisms with IAM numbers can be obtained from the Institute of Applied Microbiology, The University of Tokyo.

Microorganisms with AHU numbers are the Catalog of Cultures 4th edition (1987) published by the Japan Federation of Microbial Strains (JFCC).
, And can be obtained from the Faculty of Agriculture, Hokkaido University.

Microorganisms with DSM numbers are deutsche Sammlung von
Catalog of Strains (1 by Mikroorganismen (DSM)
983) and can be obtained from the DSM.

The medium used in the present invention is not particularly limited as long as the bacteria can grow. For example, as a carbon source, sugars such as glucose and sucrose, alcohols such as ethanol and glycerol, organic acids such as acetic acid and propionic acid,
Hydrocarbons such as paraffin, or a mixture thereof, as a nitrogen source, ammonium sulfate, yeast extract, inorganic organic nitrogen-containing compounds such as urea, other inorganic salts, trace metal salts,
It is possible to appropriately mix and use nutrient sources such as vitamins used in ordinary culture. If necessary, it is also possible to add a factor that promotes the growth of microorganisms or a substance effective for maintaining the pH of the medium. Similarly, butanediol or pentanediol can be added to the medium and cultured to enhance the oxidizing ability of the microorganism.

The culture method is such that the pH of the medium is 4.0 to 9.5 and the culture temperature is 20 to 45.
It is preferable to carry out culturing at a temperature in the range of 0 ° C for 1 to 5 days under oxygen concentration conditions suitable for growth of the microorganism.

As a method of the oxidation reaction, a diol is added to the culture solution from the beginning of the culture or during the culture, cells are separated by centrifugation or the like, and the cells are directly or washed, and then the buffer solution is added. Alternatively, there is a method in which a diol is added to a product resuspended in water or the like and reacted. In this reaction, it may be better to add a carbon source such as glucose or sucrose as an energy source. In addition, the microbial cells may be live cells, crushed cells, treated with acetone,
It may be subjected to a treatment such as freeze-drying. These cells can also be used after being immobilized on a carrier.

The diol may be added as it is, or may be dissolved in an organic solvent that does not affect the reaction or dispersed in a surfactant or the like, and added all at once from the beginning of the reaction or in divided portions.

The reaction is carried out at pH 3-9, more preferably pH 5-8, at 10
It is carried out at a temperature of -60 ° C, more preferably 20-40 ° C for 1 to 120 hours with stirring.

The concentration of the substrate is not particularly limited, but is preferably about 0.1 to 10%.

The oxyketone produced by the reaction can be easily obtained directly from the reaction solution or after separation of the bacterial cells, extraction with an organic solvent, and ordinary purification methods such as column chromatography and distillation.

(Examples) Hereinafter, the present invention will be described in detail with reference to Examples, but the present invention is not limited thereto.

The amount of oxyketone produced in the examples was determined by gas chromatography.

Example 1 Glucose 1%, yeast extract 0.3%, peptone 0.5%,
Dipotassium phosphate 0.1%, magnesium sulfate heptahydrate 0.05
%, 1,3-butanediol 0.5% (pH 7.2) in 10 ml of a medium in a test tube with a diameter of 21 mm, and after sterilization, inoculate each of the strains shown in Table 1 and reciprocate at 30 ° C for 24 hours. Shaking culture was performed. After completion of the culture, the cells were separated by centrifugation and washed once with physiological saline to obtain viable cells. 1 ml of water in this live cell
Was added to prepare a bacterial cell suspension. 1 ml of this microbial cell suspension was put into a test tube having a diameter of 21 mm, and 1 ml of a 2% (w / v) aqueous solution of 1,3-butanediol was added thereto, and the mixture was shaken reciprocally at 30 ° C. for 24 hours for reaction. After completion of the reaction, cells were removed by centrifugation, and the resulting supernatant was subjected to gas chromatography to quantify the produced 4-hydroxy-2-butanone. Table 1 shows the obtained results.

(Effects of the Invention) According to the present invention, according to the reaction using a microorganism,
It is possible to provide a method for efficiently producing a corresponding oxyketone from a diol having 4 or 5 carbon atoms under normal pressure conditions.

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Claims (1)

(57) [Claims] 1. At least one diol selected from butanediol and pentanediol is used in the genus Acinetobacter, Achrobacter.
mobacter) genus, Aeromonas genus, Agrobacterium genus, Alcaligenes (A
lkaligenes), Arthrobacter
Genus, Aureobacterium genus, Bacillus genus, Brevibacte
rium) genus, Citrobacter genus, Enterobacter genus, Escherichia (Escher)
ichia), Flavobacterium
Genus, Klebsiella genus, Lactobacillus genus, Methyloba
cterium, Micrococcus, Mycobacterium, Paracoccus
genus acoccus, genus Pimelobacter, genus Proteus, genus Serratia, genus Vibrio, genus Xanthomonas
Genus, Ambrosiozyma, Arthroascus, Botryoa
scus), Brettanomyces, Candida, Citeromyc
es), Clavispora, Cryptococcus, Debaryomes
genus yces, genus Dekkera, dipodascus (D)
ipodascus genus, Endomyces genus, Filobasidium genus, Geotricum (Ge
genus otrichum, Guilliermondell
a) genus, Hansenula genus, Hanseniaspora genus, Issatchenki
a) genus, Kluyveromyces genus, Lodderomyces genus, Lipomyces (Lypomyces)
ces genus, Metshnikowia genus, Pachsolen genus, Pichia genus, Saccharomyces genus, Saccharomyces genus
charomycodes), Schizobl
genus astosporion, Schizosacch
aromyces), Schwanniomyces
Genus, Selenozyma (Selenozyma), Selenotyra (Sele
notila), Stephanoascus
Genus, Sterigmatomyces genus, Sporidiobolus genus, Syringospora genus, Torulaspora
Genus, Torulopusis genus, Trichosporon genus, Trigonopusis
Genus, Wickerhamia genus, Wickermiella genus (Wickermiella) genus, Wingea genus (Wingea) genus, Zygosaccharomyces (Zygosaccharomyces) genus belonging to the genus of oxyketone characterized by being oxidized by a microorganism capable of oxidizing Production method.