JPH0920726A - Carbonate compound and production of 1-benzyloxy-3-chloro-2-propanol using the same compound - Google Patents

Abstract

PROBLEM TO BE SOLVED: To produce a new carbonate compound which is a precursor of benzyloxy-chloro-propanol, preservable for a long period without forming an epoxide and capable of affording the benzyloxy-chloro-propanol by reacting a specific microorganism with the carbonate compound. SOLUTION: This carbonate compound of the formula (R is an alkyl), e.g. 1-benzyloxy-3-chloro-2-methoxycarbonyloxypropane. The carbonate compound of the formula is obtained by reacting, e.g. (A) 1-benzyloxy-3-chloro-2-propanol with (B) a monoalkyl halogenated carbonate in the presence of a base in an organic solvent. A microorganism, belonging to the genus Alcaligenes, Achromobacter, Acinetobacter, Agrobacterium, Bacillus, etc., and having the ability to produce the component (A) when reacted with the compound of the formula, its cultured product or a treated microbial substance is reacted with the compound of the formula to readily produce the component (A).

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a carbonate compound and a method for producing 1-benzyloxy-3-chloro-2-propanol using the compound.

[0002]

BACKGROUND OF THE INVENTION 1-Benzyloxy-3-chloro-2-
Propanol has previously protected the terminal hydroxyl group with a benzyl group, so it is easy to introduce other functional groups in place of the chlorine atom and hydroxyl group bonded to the compound,
It is an extremely important compound as a synthetic raw material for pesticides, other physiologically active substances, and new materials such as liquid crystal materials.

However, since 1-benzyloxy-3-chloro-2-propanol has a chlorine atom and a hydroxyl group bonded to adjacent carbon atoms, if the compound is stored for a long period of time, it will remain in the air. Reacts with water and the like, often forming epoxide. In order to suppress such a phenomenon as much as possible, it is general to protect the hydroxyl group with some protective group other than a benzyl group.

Conventionally, as a method of protecting a hydroxyl group, a method of reacting with a carboxylic acid anhydride such as acetic anhydride to acylate the hydroxyl group is generally used. However, 1-benzyloxy-
When 3-chloro-2-propanol is protected with the protecting group, hydrolysis must be carried out under basic conditions when deprotecting the original 1-benzyloxy-3-chloro-2-propanol. I will have to do it. This hydrolysis further progresses, and epoxide is easily produced as a by-product, so that the protection with the acyl group or the like is extremely inconvenient for the purpose of protecting the above-mentioned hydroxyl group in order not to generate the epoxide.

[0005]

Therefore, the hydroxyl group is protected by a protecting group, and the protecting group is deprotected without producing epoxide or the like to give 1-benzyloxy-
It has been desired to develop a compound which can be used as a 1-benzyloxy-3-chloro-2-propanol precursor capable of producing 3-chloro-2-propanol.

[0006]

[Means for Solving the Problems] The inventors of the present invention have conducted extensive studies to solve such problems. As a result, 1-
A compound in which a hydroxyl group of benzyloxy-3-chloro-2-propanol is protected by an alkoxycarbonyl group is allowed to act on a specific microorganism to form 1-benzyloxy-3-chloro-2 without forming epoxide.
-It was found to be hydrolyzed to propanol, which led to the completion of the present invention.

That is, the present invention has the general formula (I)

[0008]

Embedded image

A carbonate compound represented by the formula (R is an alkyl group).

The present invention also provides the genera Alcaligenes, Achromobacter, Acinetobacter, Agrobacterium, Arthrobacter, Bacillus, Brevibacterium, Cellulomonas, Corynebacterium,
Chromobacterium, Flavobacterium, Micrococcus, Proteus, Pseudomonas, Serratia, Salmonella, Xanthomonas, Nocardia, Pseudonocardia, Rhodococcus, Candida, Cryptococcus, Debaryomyces ,
Endomyces genus, Hansenula genus, Chloequera genus, Kliberomyces genus, Pichia genus, Rhodotorula genus, Saccharomycodes genus, Saccharomycopsis genus, Schizosaccharomyces genus, Schwanniomyces genus, Sporobolomyces genus, Torula genus, Tolulopsis, Trichosporon, Brennereifefe, Octosporomyces, Bretanomyces, Geotricum, Williopsis, Bickerhamia, and 1-benzyloxy-3-chloro-2 when acted on the above carbonate compound A microorganism having the ability to produce propanol,
Alternatively, 1-benzyloxy-produced by reacting the culture solution or the treated product of the bacterial cells with the above carbonate compound.
Also provided is a method for producing 1-benzyloxy-3-chloro-2-propanol, which comprises collecting 3-chloro-2-propanol.

In the above general formula (I), R represents an alkyl group. The alkyl group may be linear or branched, and the carbon number thereof is not particularly limited. Generally, the number of carbon atoms is 1 to 3 because of easy hydrolysis by microorganisms.
Is preferably an alkyl group of Specific examples of the alkyl group suitable for the present invention include a methyl group, an ethyl group, an n-propyl group, an iso-propyl group and the like.

Specific examples of the carbonate compound represented by the general formula (I) include 1-benzyloxy-3.
-Chloro-2-methoxycarbonyloxypropane, 1
-Benzyloxy-3-chloro-2-ethoxycarbonyloxypropane, 1-benzyloxycarbonyl-3
-Chloro-2-n-propoxycarbonyloxypropane, 1-benzyloxycarbonyl-3-chloro-2-
Iso-propoxycarbonyl propane etc. can be mentioned.

The structure of the carbonate compound represented by the general formula (I) can be confirmed by the following means.

(1) ¹ H-nuclear magnetic resonance spectrum ( ¹ H-
By measuring (NMR), the bonding mode of hydrogen atoms present in the carbonate compound represented by the general formula (I) can be known.

(2) By measuring the infrared absorption spectrum (IR), the characteristic absorption derived from the functional group of the carbonate compound represented by the general formula (I) can be observed.

In the carbonate compound represented by the general formula (I), absorption based on a C = O bond can be observed in the vicinity of 1700 to 1780 cm ^-1 .

(3) Mass spectrum (MS) is measured,
A molecular ion peak (hereinafter abbreviated as M ⁺ ) of the carbonate compound represented by the general formula (I) is observed.
Therefore, the molecular weight can be determined.

(4) By subtracting the sum of the weight percentages of carbon, hydrogen, chlorine and each element from 100 by elemental analysis, the weight percentage of oxygen can be calculated and therefore the composition formula can be determined.

In the present invention, the carbonate compound represented by the general formula (I) may be produced by any method. The following method can be mentioned as a typical manufacturing method. That is, a method of reacting 1-benzyloxy-3-chloro-2-propanol with a halogenated carbonic acid monoalkyl ester in the presence of a base in an organic solvent (hereinafter abbreviated as reaction (1)) can be mentioned.

Examples of the base used in the reaction (1) include tertiary amines and alkali metal carbonates.
Specific examples thereof include triethylamine, tripropylamine, tributylamine, methyldiisopropylamine, N, N, N ', N' as tertiary amines.
-Tetramethylethylenediamine, N, N, N ', N'
-Aliphatic tertiary amines such as tetramethylpropylenediamine, pyridine, 4-dimethylaminopyridine, N, N-
Examples thereof include aromatic tertiary amines such as dimethylbenzylamine and N, N-dimethylaniline, and examples of the alkali metal carbonates include sodium carbonate and potassium carbonate. The charging molar ratio of the base to the carbonate compound represented by the general formula (I) may be appropriately determined as necessary, but it is usually 1 to 10 times mol, preferably 1 to 5 times mol. It is common.

In the reaction (1), the molar ratio of the carbonate compound represented by the general formula (I) and the halogenated carbonic acid monoalkyl ester may be appropriately determined according to need, but is usually 1 to 8 times. Mol, preferably 1-5
It is generally used in a double molar range. As the halogen of the halogenated carbonic acid monoalkyl ester, chlorine, bromine or the like is used.

In reaction (1), it is generally preferable to use an organic solvent. Examples of the solvent that can be suitably used as the solvent include dioxane, tetrahydrofuran,
Ethers such as diethyl ether and diisopropyl ether; methylene chloride, chloroform, carbon tetrachloride, 1,
Halogenated hydrocarbons such as 2-dichloroethane; Nitriles such as acetonitrile and propionitrile; N, N-
Amides such as dimethylformamide and N, N-dimethylacetamide; ketones such as acetone and methyl ethyl ketone; esters such as ethyl acetate and isopropyl acetate;
Aromatic hydrocarbons such as toluene, benzene, xylene,
Examples thereof include halogenated aromatic hydrocarbons such as chlorobenzene.

The reaction time varies depending on the compound and cannot be particularly limited, but it is preferably selected from the range of 1 to 30 hours.

The reaction temperature in the reaction (1) is not particularly limited because it varies depending on the compound, but is usually -20 to 100 ° C, preferably -10 to 80 ° C.
It is preferable to select from the range.

The method for isolating and purifying the carbonate compound of the general formula (I) obtained by the reaction (1) is not particularly limited. For example, water is added to the reaction solution to dissolve the precipitated salt, When the reaction is carried out in an organic solvent which is soluble in water, an organic solvent which is not soluble in water is added for extraction, and when the reaction is carried out in an organic solvent which is insoluble in water, a method of extracting with that solvent is mentioned. You can

After washing with water, the organic solvent is dried with a drying agent such as anhydrous sodium sulfate or anhydrous magnesium sulfate, the organic solvent is distilled off, and the residue is separated and purified by column chromatography or vacuum distillation. The target product can be obtained.

In the present invention, the carbonate compound represented by the general formula (I) thus obtained is added to the genera Alcaligenes, Achromobacter, Acinetobacter, Agrobacterium, Arthrobacter and Bacillus. , Brevibacterium, Cellulomonas, Corynebacterium, Chromobacterium, Flavobacterium, Micrococcus, Proteus, Pseudomonas, Serratia, Salmonella, Xanthomonas, Nocardia, Pseudonocardia Genus, Rhodococcus, Candida, Cryptococcus, Debaryomyces, Endomyces, Hansenula,
Cloechkela, Kliberomyces, Pichia, Rhodotorula, Saccharomycodes, Saccharomyces, Schizosaccharomyces, Schwanniomyces, Sporoboromyces, Torula, Torlopsis, Trichosporon, Belongs to the genus Brennereifefe, the genus Oktosporomyces, the genus Bretanomyces, the genus Geotricum, the genus Williopsis, the genus Bickerhamia,
When the carbonate compound is allowed to act as a substrate, a microorganism having the ability to hydrolyze its alkoxycarbonyl group, or a culture solution or a treated product of bacterial cells thereof is allowed to act.

As a result, the substrate is hydrolyzed and 1-
Benzyloxy-3-chloro-2-propanol is produced without forming epoxides and the like. Here, the above-mentioned microorganism can be used without any limitation as long as it has the above-mentioned properties. Specifically, Acinetobacter calcoaceticus (Acinetobacter calcoaceticus)
IFO 12552), Acinetobacter calcoaceticus (IFO 13006), Alcaligenes faecalisIFO 13
111), Achromobacter polymorph
polymorph AKU0122), Achromobacter superficialis AKU 012
3), Agrobacterium
rium radiobacter IAM 1526), Agrobacterium
Agrobacterium tumefaciens IFO 30
58), Arthrobact Simplex
er simplex IFO 3530), Bacillus Natto (Bacillu
s natto IFO 3335), Bacillus natto (Bacillus nat)
to IFO 3936), Bacillus roseus
IAM 1257), Bacillus subtilis
lis IFO 3007), Bacillus subtilis
tillis IFO 3026), Bacillus subtilis (Bacillus
subtillis IFO 3037), Bacillus subtilis (Bacill
us subtillis IFO 3032), Bacillus subtilis (Bac
illus subtillis IAM 1193), Bacillus subtillis var.niger IFO 302
2), Bacillus subtilis bar Atelimus (Bacil
lus subtillis var.aterrimus IFO 3038), Bacillus
Bacillus sphaericus IFO 3525, Bacillus circulans IFO 332
9), Bacillus subtilis marberg (Bacillus su
btillis marburg IFO 14144), Bacillus subtillis IFO 3007, Bacillus sphaericus IFO 3341, Bacillus sphaericus IFO 352
6), Bacillus subtillis IFO
3026), Bacillus subtillis I
FO 3134), Brevibacterium divaricatum 16
27 (Brevibqcterium divaricatum 1627 NRRL 231
1), Brevibacterium ammoniagenes (Breviba
cterium ammoniagenes IFO 12071), Celluomonas uda IFO 3747, Corynebacterium glutamicum No. 534 (Corynebacterium glut
amicum No. 534 ATCC 13032), Corynebacterium glutamicum No. 614 (Corynebacterium glutamic
um No. 614 ATCC 13060), Corynebacterium equi IAM 1038, Corynebacterium fascians IAM 1
079), Corynebacterium Paulo Metabolum (Coryn
ebacterium paurometabolum IFO 12160), Chromobacteriumiodinum IFO 3
558), Escherichia coli IFO 33
66), Flavobacterium esteroaromaticum ATCC 8091, Micrococcus luteus IFO 306
4), Proteus rettgeri IFO
13501), Pseudomonas fluorescens (Pseudom
onas fluorescens IFO 12055), Pseudomonas fluorescens IFO 3925),
Pseudomonas synxatha
IFO 3906), Pseudomonas stutzeri (Pseudom
onas stutzeri ATCC 17588), Serratia marcescens (Serratiamarcescens IFO 3054), Salmonella typhimuriumu IFO 1252
9), Xanthomonas translucence (Xanthomon
as translucens IFO 13558), Nocardia asteroides IFO3384, Nocardia autotrophica IFO 12
743), Nocardia corallina
IAM 12121), Nocardia Esislopolis (Nocardia
erythropolis IAM 12122), Nocardia erythropolis IAM 1399, Nocardia erythropolis IAM 140
0), Nocardia erythr
opolis IAM 1414), Nocardia Erythropolis (N
ocardia erythropolis IAM 1428), Nocardia erythropolis IAM 1440, Nocardia erythropolis (Nocardia erythropolis IAM 1428)
IAM 1452), Nocardia Erythropolis (Nocardia
erythropolis IAM 1463), Nocardia erythropolis IAM 1474, Nocardia erythropolis IAM 148
4), Nocardia erythr
opolis IAM 1494), Nocardia Erythropolis (N
ocardia erythropolis IAM 1503), Nocardia italica JCM 3163, Nocardia mexicana IFO 3927, Nocardia minima IAM 0374, Nocardia pseud
osporangifera IAM 0501), Nocardia Rubra (No
cardia rubra IAM 12124), Nocardia Lugosa (No
cardiarugosa JCM 3193), Nocardia species (Nocardia sp. IFO 14326), Pseudonocardia fastidiosa IF
O 14105), Rhodococcus coprofilius (Rhdococ
cus coprophilius JCM 3200), Rhodococcus corallinus JCM 3199, Rhodococcus eqi ATCC 2151, Rhodococcus eqi ATCC 6939, Rhodococcus eqi ATCC 6939, Rhodococcus eqi ATCC 6939, Rhodococcus eqi ATCC 6939 ,
Rhodococcus eqi ATCC 21690,
Rhodococcus eqi IAM 1038,
Rhodococcus erythrop
olis IFO 12539), Rhodococcus erythropolis (Rh
odococcus erythropolis IFO 12320), Rhodococcus
Rhodococcus erythropolis IFO 1253
8), Rhodococcus erythropolis
thropolis IFO 12540), Rhodococcus erythropolis JCM 3132, Rhodococcus erythropolis JCM
3201), Rhodococcus rhodococcus
hodochrous JCM 3202), Rhodococcus rocconi (R
hodococcus rhoconii JCM 3203), Rhodococcus ruber JCM 3205, Rhodococcus
Rhodococcus rubropertinctu
s JCM 3204), Rhodococcus ter
rae JCM 3206), Candida Giramon Giver Membrana Efaciens (Caidida guilliermondii va
r.membranaefaciens IFO 0643), Candida lipolytica NRRL Y-6795, Candida parapsilosis IFO 0
708), Canndida solaniIFO 07
62), Canndida maltosa I
FO 1975), Canndida kefyr
IFO 0882), Debaryomyces Hanseny (Debaryom
yces hansenii IFO 0023), Cryptococcus laurentii IFO 0609, Endomyces ovetensis IFO
1201), Hansenula henricii
IFO 1477), Kloeckera
corticis IFO 0868), Kluyveromyces lactis IFO 1090, Pichia pseudopolymorpha IFO 102
6), Rodotorula glutinis var.diarenensis IFO 041
5), Rodotorula glutinis
IFO 0389), Rodotorula
glutinis IFO 0688), Rhodotorula graminis (Rodot
orula graminis IFO 1422), Rodotorula Minuta (R
odotorula minuta IFO 0387), Rodotorula minuta IFO 0412, Rodotorula minuta bar texensis (Rodotorula minuta var.texe)
nsis IFO 0879), Rodotorula minuta var. texensis IFO 100
6), Rodotorula Minuta Bar Texensis (Rodo
torula minuta var. texensis IFO 0932), Rodotorula pallida IFO 0715, Rodotorula rubra IFO 0001, Rodotorula rubra IFO 0918, salmon Rodotorula rubra IFO 08ces IFO 0309), Saccharomycodes Ludovisi (Saccharomycodes lu
dwigii IFO 0339), Saccharomycopsis fibuligera IFO0103, Saccharomycopsis lipolytica (Saccharomycopsis l)
ipolyticaIFO 0746), Shizosaccharomyces pombe IFO 0346, Schwanniomyces occid
entalis IFO 0371), Sporobolomyces holsaticus IFO 1032, Sporobolomyces odorus IFO1
035), Sporoboromyces salmoni color bar
Polymixer (Sporobolomyces salmonicolor var.polymy
xa IFO 1039), Torrula Braber Alpha (Tor
ura rubra var.alpha IFO 0415), Torulopsis aeria IFO 0881, Trichosporon cutaneum IFO 1198, Trichosporon cutaneum IF
O 0173), Brenne Raifefe Lasse 2 (Brenne
reihefe Rasse 2 AKU 4009), Octosporomyces
Octosporomyces octosporus IFO-035
3), Brettanomyces brucellensis (Brettanom
yces bruxellensis IFO-0628), Geotrichu
m capitatum IFO-0743), Williopsis californica IFO-0800, and Wickerhamia fluorescen
s IFO 1116) and the like are preferably used.

As a medium used for culturing the above-mentioned microorganism, known medium is used. For example, glucose, sucrose, fructose, glycerol, sorbitol, molasses, soluble starch and other carbon sources, meat extract, yeast extract, polypeptone, peptone, nitrates, ammonium salts and other nitrogen sources, and potassium phosphate monobasic, There is no particular limitation as long as it contains an inorganic salt such as dibasic potassium phosphate, sodium chloride, or magnesium nitrate.

The form of the medium may be liquid or solid. The culture method may be static culture, shaking culture, or aeration-agitation culture, but liquid culture by aeration-agitation is suitable for large-scale culture. The culture temperature is 15 to 45 ° C,
Preferably, the culture is performed at 20 to 40 ° C. for usually 20 to 48 hours.

In the present invention, the above-mentioned microorganism is used as the substrate,
Alternatively, as a method of allowing the culture solution or the treated product of the bacterial cells to act, a method of acting on a substrate which is usually carried out in an enzymatic reaction utilizing a microorganism is adopted without any limitation. For example, a method in which the substrate is allowed to act by adding the substrate to the culture medium of the microorganism is mentioned. In this case, the substrate may be added to the medium from the beginning or may be added during the culture.

The substrate may be treated with the above-mentioned treated product of microorganisms or cells in an inorganic or organic solvent that does not inhibit the reaction, preferably in an aqueous solvent. In the present invention, the treated microbial cells include, for example, washed microbial cells, dried microbial cells, microbial cell grinds, autolyzed microbial cells, sonicated microbial cells, microbial cell extracts, or microbial cells. A lipase obtained by purifying a body extract or the like can be used without particular limitation. Here, as the microorganism, the cell extract, and the lipase obtained by purifying the cell extract, those immobilized by a known cell or enzyme immobilization method can be used.

In the present invention, the concentration of the substrate when the microorganism thus obtained, or its culture solution or treated product of bacterial cells is allowed to act, is not particularly limited. Usually in the range of 0.01 to 10% by weight, preferably 0.05 to 5
It may be appropriately selected from the weight percent.

Further, in the present invention, the pH of the reaction medium when the microorganism, or the culture solution or treated product of bacterial cells thereof is acted in this manner is not particularly limited, but it is usually from pH 6 to pH 6. It is preferably in the range of 10.
Furthermore, the concentration of the microorganisms and the treated cells treated when they act is
Although it cannot be determined unambiguously due to differences in the degree of purification of treated bacterial cells, it is usually 0.05-10 in terms of protein amount.
It is appropriately selected from the range of weight%. The working temperature is not particularly limited, but a range of 10 to 50 ° C, preferably 30 to 45 ° C is suitable. The action time cannot be unconditionally determined because it depends on the substrate concentration and the type of strain to be used, but it is usually sufficient to act for 3 to 80 hours.

As described above, after the hydrolysis reaction of the carbonate compound represented by the general formula (I), the produced 1-benzyloxy-3-chloro-2-propanol is collected. The method for collecting this product is not particularly limited, and for example, the reaction solution is extracted with an organic solvent such as ethyl acetate or methylene chloride, the solvent is distilled off, and the residue is easily separated by thin layer chromatography. Separation and purification can be performed.

Some 1-benzyloxy-3-propanol obtained by this reaction has high optical purity. This is because the rate of hydrolysis of the R-form and S-form of the carbonate compound represented by the general formula (I) varies depending on the strain or the like used. That is, the present invention
It is also effective as a method for producing optically active 1-benzyloxy-3-chloro-2-propanol.

[0037]

The carbonate compound represented by the general formula (I) of the present invention is 1-benzyloxy-3-chloro-2.
-Easily derived from propanol and not converted to compounds such as epoxide even when stored for a long time. In that sense, it is a derivative effective for storing 1-benzyloxy-3-chloro-2-propanol, which is a very important compound as various industrial raw materials.

The carbonate compound is a gene of Alcaligenes, Achromobacter, Acinetobacter, Agrobacterium, Arthrobacter, Bacillus, Brevibacterium, Cellulomonas, Corynebacterium, Chromobacterium. Genus, Flavobacterium, Micrococcus, Proteus, Pseudomonas, Serratia, Salmonella, Xanthomonas, Nocardia, Pseudonocardia, Rhodococcus, Candida, Cryptococcus, Debaryomyces, Endomyces , Hansenula,
Cloechkela, Kliberomyces, Pichia, Rhodotorula, Saccharomycodes, Saccharomyces, Schizosaccharomyces, Schwanniomyces, Sporoboromyces, Torula, Torlopsis, Trichosporon, Brennerifefe, Genus Octosporomyces, Brettanomyces, Geotricum, Williopsis, Bykerhamia, or microorganisms belonging to the genus Bickerhamia, or a culture solution or a treated product thereof can be easily applied without producing epoxide as a by-product. 1-Benzyloxy-3-chloro-2-propanol can be prepared.

[0039]

Hereinafter, the present invention will be described with reference to examples.
The invention is in no way limited to these examples.

Example 1 (1) Synthesis of Substrate A 4-necked flask equipped with a stirrer and a thermometer was charged with 1-benzyloxy-3-chloro-2-propanol 135.
0 g (0.67 mol), pyridine 78.1 g (0.8
(0 mol) was dissolved in 200 ml of methylene chloride and stirred at room temperature.
g (0.74 mol) was added dropwise over 2 hours so that the internal temperature was 30 ° C. or lower. After stirring for 8 hours, 200 ml of methylene chloride was added to the reaction solution and washed with 200 ml of water. Furthermore, this methylene chloride solution was added with 2m hydrochloric acid 200m
1, washed with 200 ml of water and dried over magnesium sulfate, and the solvent was distilled off. The obtained yellow transparent liquid was replaced with 8
When vacuum distillation was carried out at Torr and 147 ° C., 145.3 g (83.8%) of 1-benzyloxy-3-chloro-2-methoxycarbonyloxypropane was obtained.

As a result of measuring the infrared absorption spectrum of this product, an absorption based on a carbonyl group was obtained at 1750 cm ^-1 . The elemental analysis values are C54.98%, H5.98.
%, Cl 13.85%, and the composition formula is C ₁₂ H ₁₅ ClO ₄
C55.71 which is the calculated value for (258.70)
%, H 5.84%, Cl 13.71%.
As a result of measuring a mass spectrum, a peak corresponding to M ⁺ was shown at m / e 258.

Further, the ¹ H-nuclear magnetic resonance spectrum (δ: ppm: tetramethylsilane standard; deuterated chloroform solvent) was measured and the results were as follows.

[0043]

Embedded image

A multiplet corresponding to four protons is shown at 3.60 to 3.78 ppm, which corresponds to the methylene protons of (b) and (d). A singlet for three protons was shown at 3.80 ppm, which corresponded to the methyl proton in (a). 4.5
A singlet for three protons was shown at 6 ppm, which corresponds to the methylene proton of (e). 4.96 to 5.01 ppm
The multiplet corresponding to one proton is shown in Fig. 3 and corresponds to the methine proton in (c). A multiplet of 5 protons was shown at 7.31 ppm, which corresponds to the proton of the benzene ring in (f).

From the above results, it was revealed that the isolated product was 1-benzyloxy-3-chloro-2-methoxycarbonyloxypropane.

(2) 1-benzyloxy-3-chloro-
Production of 2-propanol The culture of the bacterium was 2.0% glucose, 1.0% sucrose, 1.0% meat extract, 0.5% peptone, 0.2% yeast extract, 0.3% sodium chloride, 0.1%. 500 ml of 4% potassium dibasic phosphate, 0.2% dibasic potassium phosphate, 0.1% magnesium sulfate heptahydrate, pH 7 to 2 ml
In addition to the L flask with a shoulder, Nocardia erythropolis IAM 1494 strain was shake-cultured at 30 ° C. for 48 hours. After culturing, the supernatant was removed by centrifugation, and 50 mM phosphate buffer (pH 7.0) 50 was added under cooling.
Washed twice with ml. 250 mg of the obtained bacterial cells was added to pH 7
Dispersed in 5 ml of 50 mM phosphate buffer solution adjusted to 3, 3-chloro-1,2-dimethoxycarbonyloxypropane was added to this so as to be 0.5% (weight / volume), and subsequently at 30 ° C., 48 The culture was shaken for an hour. After the reaction, 5 ml of ethyl acetate was added for extraction.

The extract was quantified by gas chromatography to find that it was 1-benzyloxy-3-chloro-2-
Propanol produced 18 mg (92.2% yield). The optical purity of the product was measured by high performance liquid chromatography using this extracted solution.
%, And the S form was slightly excessively produced.

Example 2 The same operations as in Example 1 were performed using the strains shown in Table 1 instead of the Nocardia erythropolis IAM 1494 strain. 1-benzyloxy-after 48-hour culture
The yield, optical purity and absolute configuration of 3-chloro-2-propanol are as shown in Table 1.

[0049]

[Table 1]

[0050]

[Table 2]

[0051]

[Table 3]

[0052]

[Table 4]

[0053]

[Table 5]

Example 3 (Synthesis of Substrate) The same operation as in Example 1 was carried out by using ethyl chlorocarbonate instead of methyl chlorocarbonate in Example 1. The obtained colorless transparent liquid was adjusted to 1.5 Torr, 16
Distillation under reduced pressure at 0 ° C. gives 1-benzyloxy-3-chloro-2-ethoxycarbonyloxypropane 16
0.8 g (88.0%) was obtained.

As a result of measuring the infrared absorption spectrum of this product, an absorption based on a carbonyl group was obtained at 1753 cm ^-1 . The elemental analysis values are C57.13%, H6.49.
%, Cl 13.11%, and the composition formula is C ₁₃ H ₁₇ ClO ₄
C57.00, which is the calculated value for (272.73)
%, H 6.28%, Cl 13.00%.

As a result of measuring the mass spectrum, m
A peak corresponding to M ⁺ was shown at / e272.

Further, the result of measurement of ¹ H-nuclear magnetic resonance spectrum (δ: ppm: tetramethylsilane standard; deuterated chloroform solvent) was as follows.

[0058]

Embedded image

A triple line corresponding to three protons is shown at 1.28 to 1.34 ppm, which corresponds to the methyl proton of (a). A multiplet of four protons was shown at 3.67 to 3.82 ppm, which corresponded to the methylene protons of (c) and (e). A quadruple line of two protons is shown at 4.17 to 4.25 ppm, which corresponds to the methylene proton of (b).
The doublet of two protons is shown at 4.56 ppm,
Corresponding to the methylene proton in (f). 4.98-5.
A multiplet for one proton was shown at 05 ppm, which corresponded to the methine proton in (d). A singlet for five protons was shown at 7.31 ppm, which corresponded to the proton of the benzene ring in (g).

From the above results, it was revealed that the isolated product was 1-benzyloxy-3-chloro-2-ethoxycarbonyloxypropane.

(2) 1-benzyloxy-3-chloro-
Production of 2-propanol 1-benzyloxy-3 instead of 1-benzyloxy-3-chloro-2-methoxycarbonyloxypropane
-Chloro-2-ethoxycarbonyloxypropane,
Nocardia Erythropolis IAM 1494 strain replaced by Corynebacterium fascias IAM
The same operation as in Example 1 was performed using 1079 strain.
When the ethyl acetate extract was quantified by gas chromatography, 15 mg (yield 73.3%) of 1-benzyloxy-3-chloro-2-propanol was produced. The optical purity of this extracted solution was measured by high performance liquid chromatography to find that it was 6.9%, and the S form was excessively produced.

Example 4 Corynebacterium fascias IAM 1079
The same operation as in Example 3 was performed using the strains shown in Table 2 instead of the strains. The yield, optical purity, and absolute configuration of 1-benzyloxy-3-chloro-2-propanol after 48 hours of culture are shown in Table 2.

[0063]

[Table 6]

[0064]

[Table 7]

[0065]

[Table 8]

Example 5 (Synthesis of Substrate) The same operation as in Example 1 was carried out by using isopropyl chlorocarbonate instead of methyl chlorocarbonate in Example 1. The obtained colorless transparent liquid is 0.1 Tor
By vacuum distillation at r, 164 ° C., 168.3 g (87.6%) of 1-benzyloxy-3-chloro-2-isopropyloxycarbonyloxypropane was obtained.

As a result of measuring the infrared absorption spectrum of this product, an absorption based on a carbonyl group was obtained at 1753 cm ^-1 . The elemental analysis values are C58.82%, H6.77
%, Cl 12.50% and a composition formula C ₁₄ H ₁₉ ClO ₄
C58.64 which is the calculated value for (286.76)
%, H 6.68%, Cl 12.36%.

As a result of measuring the mass spectrum, m
A peak corresponding to M ⁺ was shown at / e286.

Further, the result of measurement of ¹ H-nuclear magnetic resonance spectrum (δ: ppm: tetramethylsilane standard; deuterated chloroform solvent) was as follows.

[0070]

Embedded image

A double line for six protons was shown at 1.27 to 1.30 ppm, which corresponded to the methyl protons of (a) and (b). Multiple lines corresponding to four protons were shown at 3.66 to 3.80 ppm and corresponded to the methylene protons of (d) and (f). The doublet of two protons was shown at 4.54 ppm, which corresponded to the methylene proton of (g). 4.82
Shows a multiple line of two protons at ~ 5.05 ppm,
This corresponds to the methine proton of (c) and (e). 7.30
A singlet for five protons is shown in ppm, which corresponds to the proton of the benzene ring in (h).

From the above results, it was revealed that the isolated product was 1-benzyloxy-3-chloro-2-isopropyloxycarbonyloxypropane.

(2) 1-benzyloxy-3-chloro-
Production of 2-propanol 1-benzyloxy-3 instead of 1-benzyloxy-3-chloro-2-methoxycarbonyloxypropane
-Chloro-2-isopropyloxycarbonyloxypropane was added to Nocardia Erythropolis IAM 1
Instead of the 494 strain, Rhodococcus erythropolis I
The same operation as in Example 1 was performed using the FO 12538 strain.

When the ethyl acetate extract was quantified by gas chromatography, 13 mg (yield 65.4%) of 1-benzyloxy-3-chloro-2-propanol was produced. In addition, when the optical purity of the product was measured using high performance liquid chromatography of this extracted solution,
It was 7.1%, and the R form was excessively produced.

Example 6 The same procedure as in Example 5 was performed using the strains shown in Table 3 in place of the Rhodococcus erythropolis IFO 12538 strain. Yield of 1-benzyloxy-3-chloro-2-propanol after 48-hour culture, optical purity,
The absolute configuration was as shown in Table 3.

[0076]

[Table 9]

[0077]

[Table 10]

[0078]

[Table 11]

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Internal reference number FI Technical indication C12R 1:01) (C12P 7/22 C12R 1:05) (C12P 7/22 C12R 1: 025) (C12P 7/22 C12R 1:07) (C12P 7/22 C12R 1: 125) (C12P 7/22 C12R 1:09) (C12P 7/22 C12R 1:13) (C12P 7/22 C12R 1:15) (C12P 7/22 C12R 1:20) (C12P 7/22 C12R 1: 265) (C12P 7/22 C12R 1:37) (C12P 7/22 C12R 1:39) (C12P 7/22 C12R 1:38) (C12P 7/22 C12R 1: 425) (C12P 7/22 C12R 1:64) (C12P 7/22 C12R 1:73) (C12P 7/22 C12R 1:72) (C12P 7/22 C12R 1: 645) (C12P 7/22 C12R (1:84) (C12P 7/22 C12R 1:88) (C12P 7/22 C12R 1:78) (C12P 7/22 C12R 1:42)

Claims (2)

[Claims] 1. A compound represented by the following general formula (I): A carbonate compound represented by (R is an alkyl group). 2. Alcaligenes, Achromobacter,
Acinetobacter, Agrobacterium, Arthrobacter, Bacillus, Brevibacterium, Cellulomonas, Corynebacterium, Chromobacterium, Flavobacterium, Micrococcus, Proteus, Pseudomonas, Serratia Genus, Salmonella, Xanthomonas, Nocardia, Pseudonocardia, Rhodococcus, Candida, Cryptococcus, Debaryomyces, Endomyces, Hansenula, Chloequera, Kliberomyces, Pichia, Rhodotorula, Saccharo Mycodes,
Saccharomycopsis, Schizosaccharomyces, Schwanniomyces, Sporoboromyces, Torula, Tolulopsis, Trichosporone, Brennereifefe, Octosporomyces, Brettanomyces, Geotricum, Williopsis 1-benzyloxy-3-chloro-2-, which belongs to the genus Bickerhamia and is reacted with the carbonate compound according to claim 1.
A microorganism having the ability to produce propanol, or a culture solution or a treated product of the microbial cell thereof is allowed to act on the carbonate compound according to claim 1, and the produced 1-benzyloxy-3-chloro-2-propanol is collected. A method for producing 1-benzyloxy-3-chloro-2-propanol, which is characterized.