JP2003277307A - Menthol derivative, method for producing the same, and antibacterial agent or sterilizer containing the same as active ingredient - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a menthol derivative obtained by converting menthol with a fungus, Rhizoctonia solani, a method for producing the same and an antibacterial agent or a sterilizer containing the derivative as an active ingredient. <P>SOLUTION: This menthol derivative is expressed by general formula (I) (wherein, R<SP>1</SP>, R<SP>2</SP>are the same or different and are each H or an alkanoyl), and the method for producing the same and the antibacterial agent or sterilizer containing the same as the active ingredient are also provided. <P>COPYRIGHT: (C)2004,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a menthol derivative obtained by subjecting menthol to microbial conversion by a filamentous fungus Rhizoctonia solani.

[0002]

2. Description of the Related Art Attempts have been vigorously made to produce high value-added compounds by microbial conversion of inexpensive, naturally-occurring organic compounds that are available in large quantities.

For example, the natural organic compound l-menthol is
By microbial conversion with Aspergillus Species,
The fact that (-)-6-hydroxymenthol is obtained as a trace component is Phytochemistry. Vol. 30, No. 12, pp398.
1-3987, 1991.

Since l-menthol is a main component of various peppermint oils and has a refreshing aroma and taste, it is used as a food,
It is widely used as an additive in cosmetics and drug formulations. Further, l-menthol is known to have various physiological activities such as antibacterial action and bactericidal action.

R. solani (Rhizoctonia solani) is known as a filamentous fungus causing seedling blight, plant rot, stem rot, cobweb disease and the like in various plants, and is classified into 32 families (famil).
Wide host consisting of 188 species in y)
(Manibhushanrao K et al., Phytotoxic
metabolite of Rhizoctonia solani. Scientific and I
ndustrial Research 40: 602 (1981)). Also R. solani
Is at least 13 anastomosis groups:
AG), that is, AG-1 to AG-12 and AG-B
It is a complex species consisting of I.

Regarding the microbial conversion reaction of an organic compound using this R. solani, for example, hydroxylation of the meta position of an aromatic organic acid (Keisuke Komoto, Masanobu Nishimura, Isamu Hiroe: Niseki Syoho, 3
5, p94 (1969)), hydroxylation of phenylacetic acid (Keisuke Komoto, Masanobu Nishimura: Nippon Setsuho, 4, p102 (197).
5)) etc. have been reported, but l-using R. solani
There have been no reports of menthol conversion reactions.

[0007]

The present invention provides a menthol derivative obtained by microbial conversion of menthol with a filamentous fungus R. solani, a process for producing the same, and an antibacterial agent or bactericide containing the menthol derivative as an active ingredient. With the goal.

[0008]

In order to achieve the above object, the present invention has the general formula [I]:

[0009]

[Chemical 7]

The present invention provides a compound represented by the formula (wherein R ¹ and R ² are the same or different and each represents a hydrogen atom or an alkanoyl group), a process for producing the same, and an antibacterial or bactericidal agent containing the compound as an active ingredient. .

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION <6,8-Dihydroxymenthol Derivative (Compound [I])> R of Compound [I] of the Present Invention
^{When 1} or R ² is an alkanoyl group, examples of the alkanoyl include linear or branched alkanoyl having 1 to 12 carbon atoms. Preferable are linear or branched lower alkanoyl having 1 to 6 carbon atoms, and more preferable are formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, pivaloyl and the like. Of these, acetyl is particularly preferable.

Among the compounds [I] of the present invention, preferred compounds include, for example, compounds in the (1S, 3R, 4R, 6S) form.

More preferred compounds among the compounds [I] of the present invention are, for example, (1S, 3R, 4R, 6S)
-6,8-dihydroxymenthol, (1S, 3R, 4
R, 6S) -6,8-Dihydroxymenthyl 3,6-
Examples include diacetate.

<6-Dihydroxymenthol Derivative (Compound [II])> General Formula [II]:

[0015]

[Chemical 8]

(Wherein R ³ and R ⁴ are the same or different and each independently represents a hydrogen atom or an alkanoyl group), when R ³ or R ⁴ is an alkanoyl group, the alkanoyl is, for example, Examples thereof include linear or branched alkanoyl having 1 to 12 carbon atoms. Preferably, a linear or branched lower alkanoyl having 1 to 6 carbon atoms is used. More preferably, formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, pivaloyl and the like can be mentioned. Of these, acetyl is particularly preferable.

Among the compounds [II] of the present invention, preferred compounds include, for example, compounds of the (1S, 3R, 4S, 6S) form.

More preferred compounds of the compound [II] of the present invention include, for example, (1S, 3R, 4S, 6
S) -6-hydroxymenthol, (1S, 3R, 4
S, 6S) -6-Hydroxymenthyl diacetate.

<Production Method of Compounds [I] and [II]> Compounds [I] and / or [II] are menthol-forming fungi R.
It can be produced by microbial conversion with solani and optionally alkanoylating the hydroxyl groups of the product. (I) Microbial conversion of menthol by R. solani can be performed, for example, as follows.

First, as a pre-culture, a medium is prepared and sterilized, and then the medium is cooled to ingest a strain, followed by shaking culture.

As the medium, a known medium may be used, for example, Richard's medium (5% (w / v) sucrose in distilled water, 0.25% (w / v) magnesium sulfate, 1% (w / v) potassium nitrate, 0.5% (w / v) potassium dihydrogen phosphate, 0.002% (w / v) iron (III) chloride), Czapak-Dox medium (distilled water, 0.2% (w / v) sodium nitrate, 0.05% (w / v) magnesium sulfate, 0.1% (w /
v) dipotassium phosphate, 0.05% (w / v) potassium chloride, 0.00
1% (w / v) ferrous sulfate, 4% (w / v) glucose) and the like. Of these, Richards medium is preferred.

A known method may be used for the sterilization method in the medium, for example, under a pressure of about 0.01 M to 1 MPa.
It may be performed at a temperature of 0 to 150 ° C. for about 1 minute to 1 hour.

The strain used is, for example, R. solani.
AG-1 and the like, and more specifically, R. solani AG-1-I
A, R. solani AG-1-IB, R. solani AG-1-IC, R. solani
Examples include AG-1-ID. Particularly, from the viewpoint of reactivity to menthol, R. solani AG-1-IC is preferable, of which R. solani AG-1-IC F-1, R. solani AG-1-IC F-4, R.
solani AG-1-IC P-1 and the like are preferable.

The ingestion amount of the strain may be, for example, about 1 to 50 mg per 100 ml of the medium.

Known conditions may be used for the pre-culture, for example, a pressure of about 0.01 M to 1 MPa and 100
It may be performed at a temperature of up to 150 ° C. for about 1 minute to 1 hour.

Next, the separately prepared medium is sterilized under pressure, and the precultured bacterial solution is added to the medium for culturing. The culture condition is, for example, 100 to 15 under a pressure of about 0.01 M to 1 MPa.
It may be performed at a temperature of 0 ° C. for about 1 minute to 1 hour. Moreover, it is preferable to use rotary culture.

The raw material menthol is added to this and a conversion reaction is carried out. As the raw material menthol, specifically, 1-menthol is preferably used. From the viewpoint of menthol conversion efficiency, the amount of menthol added is the volume of the medium.
About 1 to 50 mg is preferable for 100 ml. The conditions of the conversion reaction are, for example, at a temperature of 25 to 27 ° C. under normal pressure, and 5 to
It only takes 10 days. The change with time of the reaction is confirmed by collecting a small amount of the bacterial solution, salting out with NaCl or the like, extracting with an organic solvent (eg, dichloromethane, chloroform), and performing TLC, GC, GC-MS analysis.

The extract obtained by the above conversion reaction is subjected to silica gel column chromatography (for example, eluent:
Compound [Ia]: by purification with (n-hexane-ethyl acetate):

[0029]

[Chemical 9]

And compound [II-a]:

[0031]

[Chemical 10]

To obtain Specifically, when 1-menthol is used as a raw material, as the compound [Ia], for example, (1S, 3R, 4R, 6S) -6,8-dihydroxymenthol is a compound [II-a]. For example, (1S, 3R, 4S, 6S) -6-hydroxymenthol is obtained. (Ii) A compound in which R ¹ and / or R ² is an alkanoyl group in the compound [I] having a hydroxyl group of the product, or a compound in which R ³ and / or R ⁴ is an alkanoyl group in the compound [II] Is produced by reacting the compound [Ia] or [II-a] obtained in (i) above with an alkanoyl halide or alkanoic acid anhydride in a solvent in the presence or absence of a base. You can

Examples of the solvent include diethyl ether, ethyl acetate, dichloromethane, chloroform and the like. Examples of the base include triethylamine,
Examples include pyridine and diisopropylethylamine. The base can also be used as a solvent. The amount of the base used may be a catalytic amount in the case of reaction with an acid anhydride,
In the case of a reaction with an alkanoyl halide, it may be, for example, about 2 to 3 equivalents with respect to the compound [Ia] or [II-a]. The reaction temperature is, for example, 50 to 120 ° C.
The reaction time may be about 5 minutes to 1 hour, for example.

Examples of the alkanoyl of the alkanoyl halide include linear or branched alkanoyl having 1 to 12 carbon atoms. Preferably, it has 1 to 6 carbon atoms.
And straight chain or branched lower alkanoyl of.
More preferably, formyl, acetyl, propionyl,
Butyryl, isobutyryl, valeryl, pivaloyl and the like can be mentioned. Of these, acetyl is particularly preferable. Examples of the halide of the alkanoyl halide include chloride and bromide.

Examples of the alkanoic acid anhydride include acetic anhydride, propanoic acid anhydride, n-butanoic acid anhydride and the like.

In the alkanoylation reaction of the compound [Ia], of the three hydroxyl groups, the tertiary hydroxyl group at the 8-position is less likely to be alkanoylated due to the effect of steric hindrance, and both of the secondary hydroxyl groups at the 3- and 6-positions or One is selectively alkanoylated.

When the compounds [I] and [II] of the present invention have an asymmetric carbon on the cyclohexane ring, a plurality of stereoisomers based on the asymmetric carbon (diastereoisomers,
Optical isomers), the present invention includes any one of these stereoisomers or a mixture thereof.

The compounds [I] and [II] of the present invention include solvates (ethanol, water, etc.) both in a free form.

<Uses of Compound [I] and Compound [II]> Compounds [I] and [II] of the present invention have a known antibacterial action against a pathogenic bacterium (snow rot: Micronectriella nivalis 305031) 8-hydroxy. The compound [I] has the same or higher bacteriolytic activity as copper quinoline.
And / or [II] can be used as an antibacterial agent or a bactericidal agent.

Specifically, the compounds [I] and / or [I
I] can be used as an antibacterial or bactericidal agent for humans, animals, and fish.

The antibacterial or bactericidal agent for humans, animals, and fishes of the present invention is pharmaceutically acceptable for parenteral administration such as injection, rectal administration, eye drop administration, oral administration in solid or liquid form, and the like. It can be formulated as a composition with a carrier.

Examples of the form of the composition of the present invention as an injection include pharmaceutically acceptable sterile water or non-aqueous solution, suspension or emulsion. Suitable non-aqueous carrier, diluent, solvent or vehicle include, for example, propylene glycol, polyethylene glycol, vegetable oils (eg olive oil) and the like. Such a composition may contain an auxiliary agent, and examples thereof include a preservative, a wetting agent, an emulsifying agent, and a dispersing agent.
These compositions can be sterilized, for example, by filtration through a bacteria-retaining filter or by mixing with sterile water immediately before use. As a formulation for eye drop administration, for example, a solubilizing agent, a preservative, an isotonicity agent, a thickener and the like may be added.

Examples of solid preparations for oral administration include capsules, tablets, pills, powders and granules. This solid dosage form comprises, for example, a compound [I] and / or [II] with at least one inert diluent (eg,
Sucrose, lactose, starch, etc.) can be mixed and prepared. This formulation also contains
In addition to the inert diluent, a lubricant (eg magnesium stearate etc.) and the like may be contained. In the case of capsules, tablets, or pills, a buffer may be included.
An enteric coating can be further applied to these solid preparations.

Liquid formulations for oral administration include inert diluents commonly used by those skilled in the art (eg, pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs containing water). Etc.) may be included. In addition to such inert diluents, auxiliary agents (eg, wetting agents, emulsifying agents, suspending agents, sweetening agents, seasoning agents, flavoring agents, etc.) can be added. Formulations for rectal administration preferably include, in addition to compound [I] and / or [II], an excipient (eg
Cocoa butter, suppository wax, etc.) may be included.

Compounds [I] and / or [II] of the present invention
Will depend on the nature of the compound administered, the route of administration, the desired treatment time, and other factors, but will generally be from about 0.1 to 1000 mg / kg per day, especially from about 0.1 to 100 mg. / Kg is preferred. If desired, this daily dose can be divided into 2 to 4 divided doses for administration.

Further, the compounds [I] and / or [II]
Can also be used as an agricultural antibacterial agent or agricultural fungicide.

The agricultural antibacterial agent or agricultural fungicide of the present invention is, for example, rice blast, sesame leaf blight, seedling blight;
It has an effect of controlling plant diseases such as red mold disease, red rust disease, yellow rust disease, black rust disease, powdery mildew disease, and black spot disease of sweet potato of wheat. In addition, for example, broad bean (rust, etc.), pea (powder, etc.), spinach (falling, etc.), celery (leaf blight, etc.), cucumber (powder, etc.), strawberry (powder, etc.) Agricultural plants such as, hops (grey mold disease, etc.), apples (red star disease, etc.), pears (black star disease, etc.), peaches (black star disease, etc.), oysters (powdery mildew, etc.), roses (powdery mildew, etc.) It is also possible to control various pathogenic bacteria of. The phytopathogenic fungi to be controlled are not limited to those exemplified above.

Compounds [I] and / or [II] of the present invention
Although it can be used as an agricultural antibacterial agent or agricultural fungicide by itself, it is preferably used as an agricultural antibacterial agent or agricultural fungicide in the form of a composition containing a known pesticide auxiliary agent. Its form is, for example, a liquid agent such as an emulsion, a wettable powder, a water solution, a suspension agent (flowable agent), an oil agent; a powder agent,
Solid preparations such as fine granules, granules, tablets and microcapsules;
Compositions in the form of fumigants; fumigants and the like are preferred. The agricultural chemicals auxiliary agent can be used, for example, for the purpose of improving the effect, stabilizing, improving the dispersibility, etc., and specifically, a carrier (diluent), a spreading agent, an emulsifying agent, a wetting agent, Examples thereof include dispersants and disintegrants.

Examples of the liquid carrier include water, alcohols such as methanol, ethanol, butanol and glycol, ketones such as acetone, sulfoxides such as dimethyl sulfoxide, methylnaphthalene, cyclohexane, animal and vegetable oils and fatty acids. You can

As the solid carrier, for example, clay, kaolin, talc, diatomaceous earth, silica, calcium carbonate, montmorillonite, bentonite, feldspar, quartz, alumina, sawdust, nitrocellulose, starch, gum arabic and the like can be used.

As the emulsifier and the dispersant, a usual surfactant can be used. For example, a spreading agent such as sodium higher alcohol sulfate, stearyl trimethyl ammonium chloride, polyoxyethylene lauryl ether; polyoxyethylene nonylphenyl. Wetting agents such as ether and dialkyl sulfosuccinate; fixing agents such as carboxymethyl cellulose and polyvinyl alcohol; disintegrating agents such as sodium lignin sulfonate and sodium lauryl sulfate can be used.

The content of the compounds [I] and / or [II] to be mixed with the agricultural antibacterial agent or agricultural bactericidal agent is not particularly limited, and it may be in the form of the agricultural antibacterial agent or agricultural bactericidal agent, and used. It can be appropriately selected according to the conditions such as purpose and method of use. For example, in powders, wettable powders, emulsifiers, etc., based on the total weight of the agricultural antibacterial agent or agricultural fungicide,
It can be selected from the range of 0.001 to 20% by weight.

The form, manufacturing method and use method of the agricultural antibacterial agent or agricultural bactericidal agent can be appropriately selected by those skilled in the art based on known methods.

The agricultural antibacterial agent or agricultural fungicide of the present invention comprises other fungicides, insecticides, acaricides, herbicides and insects in addition to the compounds [I] and / or [II] which are active ingredients. Active ingredients such as growth regulators, fertilizers and soil conditioners may be optionally mixed.

The application object and application form of the agricultural antibacterial agent or agricultural bactericidal agent of the present invention are not particularly limited, but for example, foliage treatment (liquid agent spraying, powder agent spraying, fumes, etc.), seed treatment (immersion, powder coating) , Smearing, etc.), soil treatment (dusting, irrigation, admixture, fumigation, etc.), water surface application (granulation, etc.), etc. For example, in the case of spraying foliage, 1 to 1000
A solution of about ppm may be used at an application rate of about 10 to 100 L per are, and in the case of water surface application, usually about 0.1 to 1 kg per are in a granule having an active ingredient of about 1 to 20%. If

Further, the compounds [I] and [II] of the present invention
Does not adversely affect the human body within a concentration range that can be used as an antibacterial agent or bactericide for humans, animals, and fish, or within a concentration range that can be used as an agricultural antibacterial agent or agricultural bactericide. It has low toxicity and high safety. In addition, the compounds [I] and [I of the present invention
I] are both colorless and have less odor than menthol.

[0057]

EXAMPLES Hereinafter, specific examples of the present invention (examples and experimental examples)
However, the present invention is not limited thereto.

Example (1) Preculture In a 500 ml Sakaguchi flask, 200 ml of Richards medium (Richar
d's medium) (5% (w / v in distilled water: (mass g / volume of medium
ml)) sucrose, 0.25% (w / v) magnesium sulfate, 1% (w / v
v) potassium nitrate, 0.5% (w / v) potassium dihydrogen phosphate, 0.0
02% (w / v) iron (III) chloride) was prepared and autoclaved at 121 ° C. for 15 minutes. Then, cool the medium and remove the strain from the slant.
After ingesting 30 mg, shake culture was performed at 25 ° C for 7 days. The strain was R. solani obtained from Dr. Muro Hyakumachi at Gifu University.
AG-1-IC F-1 (MAFF305909) or R. solani AG-1-IC F
-4 was used. (2) Change over time After adjusting 100 ml of Richards medium to a 200 ml flask, autoclaving is performed, and 5 ml of the precultured bacterial solution is added, and the temperature is 27 ° C.
The cells were cultivated for 7 days in rotation. After 7 days, 30 mg of a substrate, l-menthol (manufactured by Wako Pure Chemical Industries, Ltd.) was added, and then a conversion reaction was performed for 5 days. Every day, 10 ml of the bacterial solution was collected, salted out with NaCl, and the conversion product was extracted with dichloromethane.
The time course of the conversion product was traced by analyzing the extract with GC and GC-MS. Figure 1 (for strain R. solani AG-1-IC F-1) and Fig. 2 (for strain R. solani AG-1-IC F-4) show the changes with time of each component in the culture solution. Show.

Table 1 shows the total amount (%) of the product obtained in the conversion reaction.

[0060]

[Table 1]

(3) Large-scale conversion 800 ml of Richards' medium was prepared in a large-scale culture tank of 1 L and sterilized under pressure. Then, add the pre-cultured bacterial solution, and
Rotation culture was carried out for one day. After 7 days, 240 mg of l-menthol as a substrate was added, and a conversion reaction was performed for 5 days thereafter. After 5 days, the whole bacterial solution was collected, salted out with NaCl, and the conversion product was extracted with dichloromethane. (4) Isolation of conversion product The extract obtained by large-scale culture is subjected to silica gel column chromatography (eluent: n-hexane-ethyl acetate).
The following conversion product was obtained by repeating the fractionation with.

(-)-(1S, 3R, 4S, 6S) -6
- hydroxy Menthol: ^{_{crystal; [α] D 25: -42.0}} ° (CHCl 3; c0.4); HR-MS m / z: 172.
1465; EI-MS m / z (relative intensity): 154 (15), 139 (48), 121 (1
3), 111 (15), 97 (40), 95 (17), 83 (33), 69 (37), 67 (1
0), 55 (67), 43 (100); IRν _max cm ^-1 : 3264, 1450, 134
3, 1029; ¹³ C-NMR: δ15.9 (C-9), δ18.1 (C-10), δ20.9
(C-8), δ25.7 (C-7), δ32.4 (C-5), δ38.3 (C-1), δ4
2.5 (C-2), δ48.5 (C-4), δ70.6 (C-3), δ75.9 (C-6); ¹
H-NMR (500.00MHz, CDCl ₃ , TMS internal standard) δppm: 0.82 (3
H, d, J = 7.0Hz), 0.99 (3H, d, J = 7.1Hz), 1.03 (3H, d,
J = 7.2Hz), 1.09 (1H, ddd, J = 12.9, 11.3, 11.2Hz), 1.1
2 (1H, ddd, J = 13.0, 11.1, 10.6Hz), 1.26 (1H, brs),
1.50 (1H, dddd, J = 11.3, 10.5, 4.2, 3.0Hz), 1.61 (1H,
dddd, J = 11.1, 11.0, 7.0, 4.0Hz), 1.76 (1H, ddd, J =
12.9, 4.2, 3.9Hz), 1.96 (1H, ddd, J = 13.0, 4.0, 3.9H
z), 2.04 (1H, brs), 2.15 (1H, ddd, J = 7.2, 7.1, 3.0H
z), 3.19 (1H, ddd, J = 11.2, 11.0, 3.9Hz), 3.45 (1H, d
dd, J = 10.6, 10.5, 3.8Hz).

(+)-(1S, 3R, 4R, 6S)-
6,8-dihydroxy-Menthol: ^{_{Oil; [α] D 25: +}} 5.2 ° (CHCl 3; c1.0); HR-MS m / z; 18
8.1413; EI-MS m / z (relative intensity): 173 (1), 155 (1), 137
(3), 116 (22), 94 (80), 83 (10), 79 (30), 70 (11), 58 (1
00), 53 (4), 43 (47); IRν _max cm ^-1 : 3332, 1369, 1159,
1022; ¹³ C-NMR: δ17.9 (C-7), δ23.7 (C-9), δ30.1 (C-
10), δ36.1 (C-5), δ38.1 (C-1), δ42.2 (C-2), δ51.8
(C-4), δ72.0 (C-3), δ74.8 (C-8), δ75.6 (C-6); ¹ HN
MR (500.00MHz, CDCl ₃ , TMS internal standard) δppm: 0.94 (1H,
ddd, J = 13.1, 10.5, 10.4Hz), 1.01 (1H, brs), 1.02 (3
H, d, J = 7.0Hz), 1.08 (1H, brs), 1.16 (1H, ddd, J = 13.
0, 4.1, 4.0Hz), 1.22 (3H, s), 1.24 (3H, s), 1.41 (1H,
dddd, J = 10.2, 10.0, 7.0, 4.0Hz), 1.56 (1H, ddd, J = 1
0.4, 10.3, 4.2Hz), 1.88 (1H, ddd, J = 13.1, 4.3, 4.2H
z), 1.94 (1H, ddd, J = 13.0, 10.1, 10.0Hz), 2.03 (1H,
brs), 3.21 (1H, ddd, J = 10.5, 10.2, 4.3Hz), 3.77 (1H,
ddd, J = 10.3, 10.1, 4.1Hz). (5) Acetylation reaction (-)-(1S, 3R, 4S, 6 obtained in (4) above.
S) -6-Hydroxymenthol 30 mg in ethyl acetate
28 ml of pyridine and 27 mg of acetyl chloride were added to 30 ml at room temperature, and the mixture was stirred at room temperature for 1 hour. Water was added to the reaction solution, washed with 5% hydrochloric acid, 5% aqueous sodium hydrogen carbonate solution and brine, dried over magnesium sulfate, and the solvent was evaporated under reduced pressure. By purifying the residue by silica gel column chromatography (eluent: n-hexane: ethyl acetate = 1: 1),
44.5 mg of (-)-(1S, 3R, 4S, 6S) -6-hydroxymenthyl diacetate was obtained. Oil; [α] _D ²⁵ : -82.2 ° (CHCl ₃ ; c1.0); EI-MS m / z (relative intensity): 154 (3), 136 (30), 121 (21), 107 (5) , 93 (40),
81 (8), 69 (8), 55 (10), 43 (100); IRν _max cm ^-1 : 2359,
2340, 1736, 1457, 1370, 1242, 1026; ¹³ C-NMR: δ 16.2
(C-9), δ17.8 (C-10), δ20.5 (C-7), δ21.1 and δ21.2
(CH ₃ CO), δ26.2 (C-8), δ29.0 (C-5), δ35.4 (C-1), δ
38.1 (C-2), δ45.2 (C-4), δ77.1 (C-6), δ170.6 and δ
170.8 (CH ₃ CO); ¹ H-NMR (500.00MHz, CDCl ₃ , TMS internal standard) δppm: 0.75 (3H, d, J = 7.1Hz), 0.85 (3H, d, J = 7.0H
z), 0.91 (3H, d, J = 7.2Hz), 1.11 (1H, ddd, J = 12.9, 1
1.3, 11.2Hz), 1.16 (1H, ddd, J = 13.0, 11.1, 10.6Hz),
1.61 (1H, dddd, J = 11.3, 10.5, 4.2, 3.0Hz), 1.67 (1H,
dddd, J = 11.1, 11.0, 7.0, 4.0Hz), 1.86 (1H, ddd, J =
7.2, 7.1, 3.0Hz), 1.92 (1H, ddd, J = 12.9, 4.2, 3.9H
z), 2.02 (1H, ddd, J = 13.0, 4.0, 3.9Hz), 2.04 (3H,
s), 2.06 (3H, s), 4.45 (1H, ddd, J = 11.2, 11.0, 3.9H
z), 4.67 (1H, ddd, J = 10.6, 10.5, 3.8Hz).

(+)-(1S, 3 obtained in (4) above
R, 4R, 6S) -6,8-Dihydroxymenthol 30
By treating mg in the same manner as above, (-)-(1
S, 3R, 4R, 6S) -6,8-Dihydroxymenthyl 3,6-diacetate 43.4 mg was obtained.

Oil; [α] _D ²⁵ : -35.8 ° (CHCl ₃ ; c1.0); E
I-MS m / z (relative intensity): 197 (1), 152 (2), 138 (1), 134 (6),
109 (3), 100 (2), 94 (77), 79 (29), 69 (3), 59 (21), 55
(7), 43 (100); IRν _max cm ^-1 : 3472, 1737, 1457, 1371,
1242, 1114, 1026, 757; ¹³ C-NMR: δ17.7 (C-7), δ21.1
And δ21.5 (CH ₃ CO), δ22.7 (C-9), δ28.3 (C-10), δ35.
3 (C-1), δ36.1 (C-5), δ38.1 (C-2), δ49.6 (C-4), δ7
2.5 (C-3), δ74.1 (C-8), 76.5 (C-6), δ170.0 and δ17
0.7 (CH ₃ CO); ¹ H-NMR (500.00MHz, CDCl ₃ , TMS internal standard)
δppm: 0.91 (3H, d, J = 7.0Hz), 1.12 (1H, ddd, J = 13.1,
10.5, 10.4Hz), 1.15 (3H, s), 1.16 (3H, s), 2.06 (3H,
s), 2.07 (1H, s), 2.11 (1H, ddd, J = 13.1, 4.3, 4.2H
z), 2.16 (1H, ddd, J = 13.0, 10.1, 10.0Hz), 2.52 (1H,
brs), 4.46 (1H, ddd, J = 10.5, 10.2, 4.3Hz), 4.84 (1H,
dddd, J = 10.3, 10.1, 4.1Hz).

Experimental example (bacteriolytic activity test) The pathogenic fungus (snow rot: Micronectriella ni) to be tested
valis 305031) in advance at 25 ° C for 7 days on PDA
(Potato dextrose agar) Extend on medium. The tip of the extended pathogen is punched out with a cork borer with a diameter of 3 mm, and 7 mm square cellophane is spread on 2% glucose agar medium, and half of the agar-containing agar is placed on the cellophane (see Fig. 3). ). Then, the hyphae are allowed to grow on cellophane at 25 ° C. for 1 to 2 days (to the extent that the hyphae are not removed from cellophane).

300 μL of sterilized water having a adjusted sample concentration was placed in a multi-dish, and cellophane with hyphae extended was floated in sterilized water and allowed to stand at 3 ° C. for 48 hours. After that, take out the cellophane on the slide glass and
Stain the cytoplasm of mycelium with 5% Cotton Blue and observe with an optical microscope (see Fig. 4).

The evaluation value of the lytic activity was determined as a lytic rate by applying the ratios of the three types of hyphae shown in FIG. 4 (no lysis, incomplete lysis, complete lysis) to the following formula.

[0069]

[Formula 1]

The results of the lytic activity test are shown in Table 2 below.

[0071]

[Table 2]

The compounds ((-)-(1S, 3R, 4S, 6S) -6-hydroxymenthol, (-)-(1S, 3R, 4S, 6S) obtained in the above Examples, which are specific examples of the present invention, ) -6-Hydroxymenthol diacetate, (+)-
(1S, 3R, 4R, 6S) -6,8-Jeydroxymenthol, (-)-(1S, 3R, 4R, 6S)
It was found that -6,8-Dychtoloxymenthol 3,6-diacetate has a lytic activity equivalent to or higher than that of copper 8-hydroxyquinoline against a pathogenic bacterium (snow rot: Micronectriella nivalis 305031).

[0073]

By converting l-menthol using R. solani, (-)-(1S, 3R, 4S, 6
S) -6-hydroxymenthol and (+)-(1S,
A method for efficiently producing 3R, 4R, 6S) -6,8-dihydroxymenthol has been found.

Compounds [I] and [II] obtained from l-menthol by the above bioconversion reaction and optionally obtained by alkanoylating the hydroxyl group of the product have high bacteriolytic activity and low toxicity. Therefore, it can be used as an antibacterial agent or a bactericidal agent.

[Brief description of drawings]

FIG. 1 is a view showing a time course of the relative concentration of a compound produced by microbial conversion of 1-menthol by R. solani AG-1-IC F-1.

FIG. 2 is a diagram showing a time course of the relative concentration of a compound produced by microbial conversion of 1-menthol by R. solani AG-1-IC F-4.

FIG. 3 is a schematic diagram of a 2% glucose elementary agar medium in a lytic activity test.

FIG. 4 is an observation view under an optical microscope when the cytoplasm of hyphae is stained with 0.05% Cotton Blue.

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61P 31/04 A61P 31/04 C07C 69/013 C07C 69/013 B 69/18 69/18 C12P 7/02 C12P 7/02 // (C12P 7/02 C12R 1: 645 C12R 1: 645) C07M 7:00 C07M 7:00 F term (reference) 4B064 AC13 CA05 CE10 DA01 DA11 4C206 AA01 AA02 AA03 AA04 CA13 DB04 DB56 MA01 MA04 MA37 MA42 MA43 MA51 MA55 MA56 MA57 MA61 MA63 MA72 MA78 MA80 MA86 NA14 ZB35 4H006 AA01 AA03 AB03 AB29 BT14 FC22 FE11 FE12 4H011 AA01 AA02 AA03 BA01 BB03 BB06

Claims (10)

[Claims] 1. A compound represented by the general formula [I]: (In the formula, R ¹ and R ² are the same or different and each represents a hydrogen atom or an alkanoyl group). 2. The compound according to claim 1, which is a (1S, 3R, 4R, 6S) body. 3. The method for producing a compound according to claim 1, wherein menthol is subjected to microbial conversion with Rhizoctonia solani, and the hydroxyl group of the product is optionally alkanoylated. 4. The menthol is 1-menthol, Rh
The process according to claim 3, wherein izoctonia solani is Rhizoctonia solani AG-1-IC. 5. A compound represented by the general formula [I]: (In the formula, R ¹ and R ² are the same or different and each represents a hydrogen atom or an alkanoyl group) An antibacterial agent or a bactericidal agent comprising the compound as an active ingredient. 6. A compound represented by the general formula [I]: (In the formula, R ¹ and R ² are the same or different and each represents a hydrogen atom or an alkanoyl group) An agricultural antibacterial agent or agricultural fungicide containing the compound as an active ingredient. 7. A general formula [II]: wherein menthol is subjected to microbial conversion with Rhizoctonia solani, and the hydroxyl group of the product is optionally alkanoylated. (In the formula, R ³ and R ⁴ are the same or different and each represents a hydrogen atom or an alkanoyl group). 8. The menthol is 1-menthol, Rh
The method according to claim 7, wherein izoctonia solani is Rhizoctonia solani AG-1-IC. 9. A compound represented by the general formula [II]: (In the formula, R ³ and R ⁴ are the same or different and each represents a hydrogen atom or an alkanoyl group) An antibacterial agent or a bactericidal agent comprising the compound as an active ingredient. 10. A compound represented by the general formula [II]: (In the formula, R ³ and R ⁴ are the same or different and each represent a hydrogen atom or an alkanoyl group) An agricultural antibacterial agent or agricultural fungicide containing an active ingredient as a compound.