WO2004003213A1 - ラクトン類の製造方法 - Google Patents
ラクトン類の製造方法 Download PDFInfo
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- WO2004003213A1 WO2004003213A1 PCT/JP2003/008217 JP0308217W WO2004003213A1 WO 2004003213 A1 WO2004003213 A1 WO 2004003213A1 JP 0308217 W JP0308217 W JP 0308217W WO 2004003213 A1 WO2004003213 A1 WO 2004003213A1
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- hydroxy fatty
- fatty acid
- decalactone
- acid
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/165—Yeast isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/04—Oxygen as only ring hetero atoms containing a five-membered hetero ring, e.g. griseofulvin, vitamin C
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/02—Oxygen as only ring hetero atoms
- C12P17/06—Oxygen as only ring hetero atoms containing a six-membered hetero ring, e.g. fluorescein
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
- C12R2001/72—Candida
Definitions
- the present invention relates to a method for producing ratatones, which are useful as fragrances and pharmaceutical intermediates, using microorganisms. Background technology
- Fragrances are roughly classified into two types depending on their raw materials or manufacturing method: flavors of chemically synthesized products (so-called “synthetic flavors”) and flavors other than chemically synthesized products (so-called “natural flavors”). And consumers in recent years tend to prefer “natural products” to avoid “synthetic products”.
- the optically active substances ⁇ -decalactone (R- ⁇ -decalactone, S- ⁇ -decalactone) and ⁇ -decalactone which are important substances in constituting natural food flavors, are natural products. Due to the very small amount present in water, it is not advantageous from a technical and economic point of view to separate them by extraction or other operations with high optical purity. Therefore, at present, “synthetic products” are generally supplied inexpensively and in large quantities, while “natural products” are small-scale and expensive.
- natural R- ⁇ -decalactone is derived from natural product raw materials or their decomposition products using only biological and physical methods without using any chemical method. Fermentation methods using microorganisms are attracting attention as a method for producing yeast.
- a method for producing ⁇ -decalactone using a microorganism using castor oil (caster oil) or a hydrolysis product thereof as a raw material is disclosed in Japanese Patent Application Laid-Open No. Sho 59-82090, Aspergillus oryzae ( Aspergillus oryzae, Candida gosa, Geotrichum klebanni i, Yarrowia lipolytics, etc.
- Aspergillus oryzae Aspergillus oryzae, Candida gosa, Geotrichum klebanni i, Yarrowia lipolytics, etc.
- There is disclosed a method for producing y-decalactone by producing decanoic acid, making it acidic by adding hydrochloric acid and the like, and then heating it to ratatonization is disclosed in Japanese Patent Publication No. 63-56295 and a report by KA MAUME et al.
- a method for producing ⁇ _decalactone from castor oil or ricinoleic acid via hydroxydecanoic acid using such microorganisms is disclosed. Furthermore, prior to these technologies, S. Okui et al., Using several strains belonging to the genus Candida, present two-hydroxydecanoic acid and y-decalactone as intermediates during the oxidative degradation of ricinoleic acid. [J. Biochem., Vol. 54, No. 6, 536-540 (1963)]. In addition, European Patent Publication 9 9 7 5 33 3 discloses a method for producing ⁇ -decalactone at 12 g / L from castor oil using Yarroia lipolytica.
- ⁇ -hydroxydecanoic acid and ⁇ -decalactone can be efficiently used without using an emulsifier or a ⁇ ⁇ regulator and adding a high concentration of castor oil and ⁇ or a hydrolysis product of castor oil as raw materials.
- emulsifier or a ⁇ ⁇ regulator and adding a high concentration of castor oil and ⁇ or a hydrolysis product of castor oil as raw materials.
- R- ⁇ -decalactone can also be produced from a racemic mixture obtained by extraction from natural products by a method of selectively separating R- ⁇ -decalactone by a method known to those skilled in the art.
- the amount of R-y-decalactone present in natural products is extremely small, and it is physically difficult to separate R-decalactone from other volatile compounds. Extracting from objects is not an economical method. Therefore, in order to meet the high demand for natural compounds as described above, an efficient natural R- ⁇ -decalactone is obtained by a method different from the chemical synthesis method of R- ⁇ -decalactone or the separation method from a racemic mixture. The development of a manufacturing method was required.
- Saccharomyces cerevisiae utilizes the reducing power of natural 2-decene-1,5-olide to 5-decanolate.
- the compound is produced by biohydrogenation from a substrate containing the corresponding unsaturated lactone, delta-decene-1-olide, delta-decene-12-lide, or a mixture thereof.
- the substance conversion method using the reducing ability of yeast requires a high substrate concentration.
- problems such as the difficulty in the action of the target, and the fact that it takes a long time to obtain the target substance. Disclosure of the invention
- the present invention has been made in view of the above-mentioned problems, and has been made using natural microorganisms, such as optically active lactones such as optically active ⁇ -decalactone and optically active ⁇ -decalactone, using microorganisms. It is intended to provide an efficient manufacturing method.
- natural microorganisms such as optically active lactones such as optically active ⁇ -decalactone and optically active ⁇ -decalactone
- the present inventors can accumulate ⁇ / -hydroxydecanoic acid at a high concentration in a culture medium using castor oil and / or a hydrolyzed product of castor oil as a carbon source.
- Microorganisms were extensively searched from known strains and nature.
- Candida sorbophila no emulsifier or pH regulator was used, and it was selected from the group consisting of castor oil, hydrolyzed products of castor oil, ricinoleic acid, and rescuelaric acid.
- Candida sorbophila is cultured in a medium containing at least one selected from the group consisting of a hydroxy fatty acid, a hydroxy fatty acid derivative, and a hydrolysis product of a hydroxy fatty acid derivative, A method for producing lactones, comprising collecting the produced lactones from the medium.
- Candida sorbophi la Candida sorbophi la ATCC74362, Candida sorbophi la ATCC 60130, Candida sorbophi la and Candi da sorbophila IF01583 The method according to [1] or [2], wherein the method is at least one selected from the group consisting of Candida sorbophi la FC58 strain deposited under No. FERM BP-8388.
- Lactones have the general formula (1)
- ring A represents a ratatone ring
- R 1 represents a hydrogen atom, a hydrocarbon group, a substituted hydrocarbon group, a heterocyclic group or a substituted heterocyclic group
- R 2 represents a hydrogen atom, a hydrocarbon group or Represents a substituted hydrocarbon group
- the ring A and R 2 may combine to form a ring.
- R 1 represents a hydrogen atom, a hydrocarbon group, a substituted hydrocarbon group, a heterocyclic group or a substituted heterocyclic group
- R 2 represents a hydrogen atom, a hydrocarbon group or a substituted hydrocarbon group
- R 3 Represents a divalent hydrocarbon group which may have a substituent having a carbon chain of 4 or more, wherein R 2 and R 3 may combine to form a ring.
- R 1 represents a hydrogen atom, a hydrocarbon group, a substituted hydrocarbon group, a heterocyclic group or a substituted hetero group
- R 2 represents a hydrogen atom, a hydrocarbon group or a substituted hydrocarbon group
- R 3 Represents a divalent hydrocarbon group which may have a substituent having a carbon chain of 4 or more
- R 4 represents an alkyl group, even if R 2 and R 3 combine to form a ring
- R 6- : R 8 are each independently a hydrogen atom or a general formula (6)
- R 1 represents a hydrogen atom, a hydrocarbon group, a substituted hydrocarbon group, a heterocyclic group or a substituted heterocyclic group
- R 2 represents a hydrogen atom, a hydrocarbon group or a substituted hydrocarbon group
- 3 represents a divalent hydrocarbon group which may have a substituent having 4 or more carbon chains
- R 4 represents an alkyl group, wherein R 2 and R 3 are bonded to form a ring.
- a group represented by at least one of R 6 to R S is a group represented by the general formula (6).
- Lactones are ⁇ -decalactone, ⁇ -valerolactone, y-xalactone, ⁇ -heptalataton, octalactone, 7-nonalactone, ⁇ -dendecalactone, ⁇ -dodecalactone, ⁇ -tetradecalactone, ⁇ -decalactone, ⁇ xactalactone, ⁇ -heptalactone, ⁇ -octalactone, ⁇ -nonalactone, ⁇ onedecalactone, eighteendedecalactone ⁇ -tridecalactone and ⁇ -tetradecalactone
- Candida sorbophila is cultured in a medium containing at least one selected from the group consisting of castor oil, a hydrolysis product of castor oil, ricinoleic acid, and rescuelaric acid. And collecting the produced ⁇ -decalactone from the medium. —A method for producing decalactone.
- Candida sorbophi la is cultured in a medium containing at least one selected from the group consisting of castor oil, a hydrolyzate of castor oil, ricinoleic acid, and rescuelaric acid.
- a method for producing ⁇ -decalactone which comprises ratating ⁇ -hydroxydoxycarboxylic acid produced in the medium.
- At least one selected from the group consisting of castor oil, a hydrolyzed product of castor oil, ricinoleic acid and rescuelaric acid is a hydrolyzed product of castor oil and Z or castor oil.
- Candida and Candida sorbophi la are cultured in a medium containing 11-hydroxypalmitic acid and ethyl or 11-hydroxypanolemitic acid ethyl ester, and the produced ⁇ -decalactone is produced.
- a method for producing ⁇ -decalactone comprising collecting from the medium.
- Candida sorbophi la is cultured in a medium containing 11-hydroxypalmitic acid and / or ethyl 11-hydroxypanolemitic acid, and is produced in the medium.
- ⁇ A process for producing ⁇ -zachalactone, which comprises rattling hydroxydecanoic acid.
- Candida sorbophi la FERM BP-8388 strain The production of ratatones in the present invention is carried out in a medium containing at least one selected from the group consisting of a hydroxy fatty acid, a hydroxy fatty acid derivative, and a hydrolysis product of a hydroxy fatty acid derivative. (Candida sorbophi la) to produce ratatones, and the ratatones are searched for from the medium.
- the production of ratatones in the present invention can also be carried out in a medium containing at least one selected from the group consisting of hydroxy fatty acids, hydroxy fatty acid derivatives, and hydrolysis products of hydroxy fatty acid derivatives.
- Candida sorbophila used in the present invention include Candida sorbophila FC58 strain, Candida sorbophila ATCC74362 strain, Candida sorbophila ATCC60 * and Candida sorbophila ATCC60 *.
- the Candida sorbophila FC58 strain dated June 10, 2010 (the date of deposit of the original material), was deposited at the National Institute of Advanced Industrial Science and Technology (AIST) at the Patent Organism Depositary Center (Tsukuba East 1-chome, Tsukuba, Ibaraki, Japan) Deposit No. FERM BP-8388 at No. 1 No. 1 Central No. 6), requesting transfer of this deposit from Hara Deposit to a deposit based on the Budapest Treaty on May 28, 2003 Has been received.
- AIST National Institute of Advanced Industrial Science and Technology
- Candida sorbophila FC58 strain was isolated from common soil in Kanagawa Prefecture according to a conventional method, and mycological properties were identified, and those properties were referred to in a taxonomy [Kurtzman, CP Barnett, JA et al., "Yeasts ⁇ 'Characteristics and identification” 3rd ed], Candida. Sorbofilah (The Yeasts, A Taxo Painting Study 4th edition (1998) Elsevier Science BV; Candida sorbophila). Therefore, the microorganism isolated from the natural world was named Candida sorbophila strain FC58 (hereinafter abbreviated as “FC58 strain”).
- FC58 strain The bacteriological properties of the FC58 strain that can be suitably used in the present invention are as follows.
- Morphology Spherical to ovoid, multiplying by multipolar budding. Formation of pseudohyphae No ascospore formation was observed during growth on Dams, Gorodokoba, malt, YM, V-8, and potato dextrose media.
- Vitamin requirement No growth on a vitamin-deficient medium is observed. Require biotin, pyridoxine, and thiamine.
- ATCC and IF0 are abbreviations of “American Type Culture Collection and Institution for Fermentation, Osaka, Japan (Fermentation Research Institute, Inc.)”, respectively, and are numbers with “ATCC” or “IF0” appended. Represents the catalog number of each strain. Each of the above Candida sorbophi la, indicated by the number with “ATCC” or “IF0”, was obtained from strains stored by the respective institutions based on these catalog numbers. , Can be used.
- Candida sorbophila used in the present invention is cultured in a medium containing hydroxy fatty acid, a hydroxy fatty acid derivative, and a hydrolysis product of Z or a hydroxy fatty acid derivative. It is possible to produce precursor hydroxy fatty acids, or to produce ratatones by ratating the lactone precursor hydroxy fatty acids, and accumulate them in the medium.
- Candida sorbophi la used in the present invention When cultured in a medium containing a hydroxy fatty acid derivative under appropriate culture conditions, etc., hydrolyzes the hydroxy fatty acid derivative, and then oxidizes the hydrolysis product to form the lactone precursor hydroxy. Produces fatty acids and accumulates them in the medium.
- the desired lactones can be obtained by lactonizing the ratatone precursor hydroxy fatty acid produced and accumulated in the medium
- Candida sorbophila used in the present invention is For example, when cultured in a medium containing castor oil, the castor oil can be hydrolyzed, and the hydrolysis product of the castor oil can be oxidized to obtain ⁇ -hydroxydecanoic acid and / or ⁇ -hydroxydecanoic acid. Decalactone can be produced and accumulated in the medium
- a medium for producing lactones at least one selected from the group consisting of hydroxyfatty acids, hydroxyfatty acid derivatives, and hydrolysis products of hydroxyfatty acid derivatives is used.
- the hydrolysis product of the hydroxy fatty acid or the hydroxy fatty acid derivative used in the present invention is not particularly limited as long as Candida sorbophila can oxidize it to produce ratatone precursor hydroxy fatty acid. It is not done.
- the hydroxy fatty acid derivative used in the present invention is obtained by hydrolyzing the hydroxy fatty acid derivative by Candida sorbophila (Candida sorbophi la), and converting the hydrolysis product into a ratatonic precursor hydroxy fatty acid by / 3 oxidation. There is no particular limitation as long as it can be produced.
- the hydroxy fatty acid used in the present invention has a hydroxyl group at least at position 6, preferably 6 to 20, counted from the carbon atom of the carboxyl group, has 6 or more carbon atoms, preferably 6 to 25, more preferably Hydroxy fatty acids having 6 to 20 are preferred.
- hydroxy fatty acid examples include, for example, the general formula (2)
- R 1 represents a hydrogen atom, a hydrocarbon group, a substituted hydrocarbon group, a heterocyclic group or a substituted heterocyclic group
- R 2 represents a hydrogen atom, a hydrocarbon group or a substituted hydrocarbon group
- R 3 Represents a divalent hydrocarbon group which may have a substituent having a carbon chain of 4 or more, wherein R 1 and R 2 or R 2 and R 3 may combine to form a ring.
- hydrocarbon group examples include an alkyl group, an alkynole group having an unsaturated bond, an aryl group, an aralkyl group and the like.
- the alkyl group may be linear, branched or cyclic, for example, having 1 or more carbon atoms, preferably 120 carbon atoms, more preferably 115 carbon atoms, and still more preferably 11 carbon atoms. And an alkyl group of 0.
- Specific examples thereof include methynole, ethyl, n-propyl, 2-propyl, n_butyl, 2-butynole, isobutynole, tert-butyl, n-pentyl, and 2-pentynole Tert-pentynole, 2-methylbutyl, 3-methynolebutynole, 2,2-dimethynolepropyl, n-xyl, 2-xynole, 3_xyl, tert-xyl, 2-methylpentyl, 3-methyl Pentyl group, 4-methynol-pentinole group, 2-methylpentane-3-yl group, n-butyl group, n-octynole group, n-noel group, n-decinole group, cyclopropyl group, cyclopentinole group, cyclopentyl group And a cyclohexy
- alkyl group having an unsaturated bond examples include an alkyl group having at least one unsaturated bond such as a double bond in a chain of the alkyl group.
- Specific examples thereof include an alkenyl group, an alkadieel group, Alkatrienyl groups and the like.
- the alkenyl group has one double bond in the chain of the alkyl group, may be linear, branched, or cyclic, and has, for example, 2 or more carbon atoms, preferably 2 20 carbon atoms, and more preferably 2 carbon atoms.
- An alkenyl group having 2 15 carbon atoms, more preferably 2 10 carbon atoms may be mentioned.
- Specific examples include ethenyl, probenenole, butenyl, pentenyl, hexenyl, otatur, nonanyl, and decenyl. And the like.
- the alkadienyl group has two double bonds in the chain of the alkyl group, may be linear, branched or cyclic, for example, has 4 or more carbon atoms, preferably 4 to 20 carbon atoms, more preferably Is an alkadiene group having 4 to 15 carbon atoms, more preferably 4 to 10 carbon atoms. Specific examples thereof include a 1,3-butadienyl group, a 2,4-butadienyl group, and a 2,3-dimethyl-11,3 butadienyl group.
- Examples of the aryl group include an aryl group having 6 to 14 carbon atoms, and specific examples include a phenyl group, a naphthyl group, an anthryl group, and a biphenyl group.
- Examples of the aralkyl group include groups in which at least one hydrogen atom of the alkyl group is substituted with the aryl group. For example, an aralkyl group having 7 to 12 carbon atoms is preferable. And a 2-phenyl / reethyl group, a 1-phenylpropyl group and a 3-naphthylpropyl group.
- heterocyclic group examples include an aliphatic heterocyclic group and an aromatic heterocyclic group.
- aliphatic heterocyclic group examples include, for example, a heteroatom having 2 to 14 carbon atoms and at least one heteroatom, preferably 1 to 3 heteroatoms such as a nitrogen atom, an oxygen atom, and a sulfur atom.
- aliphatic heterocyclic group examples include, for example, a pyrrolidyl-2-one group, a piperidino group, a piperazyl group, a morpholino group, a tetrahydrofuryl group, a tetrahydroviranyl group, and the like.
- aromatic heterocyclic group examples include a heteroatom having 2 to 15 carbon atoms and containing at least one, preferably 1 to 3 heteroatoms such as a nitrogen atom, an oxygen atom, and a sulfur atom.
- substituted hydrocarbon group examples include a substituted alkyl group, an alkyl group having a substituted unsaturated bond, a substituted aryl group, a substituted aralkyl group, and the like, in which at least one hydrogen atom of the above hydrocarbon group is substituted with a substituent. No.
- substituents examples include a hydrocarbon group, a substituted hydrocarbon group, a heterocyclic group, a heterocyclic group, an alkoxy group, an aryloxy group, an aralkyloxy group, an anolexoxycarbonyl group, an aryloxycarbonyl group, and an aralkyl group.
- the group represented by R 1 is preferably an alkyl group or an alkyl group having an unsaturated bond.
- Examples of the divalent hydrocarbon group having 4 or more carbon chains which may have a substituent include a divalent hydrocarbon group having 4 or more carbon chains and a divalent substituted hydrocarbon group having 4 or more carbon chains.
- Examples of the divalent hydrocarbon group having 4 or more carbon chains include an alkylene group, an alkylene group having an unsaturated bond, a divalent aromatic group, and the like.
- the alkylene group having 4 or more carbon chains may have at least 4 carbon chains, preferably 4 to 20 carbon atoms, more preferably 4 to 15 carbon chains, and may be linear, branched or cyclic. And an alkylene group. Specific examples thereof include a tetramethylene group, a pentamethylene group, a hexamethylene group, a heptamethylene group, an octamethylene group, a nonamethylene group, a decamethylene group, and one (CH 2 ) m —o—C 6 .
- alkylene group having an unsaturated bond of 4 or more carbon chains examples include alkylene groups of 4 or more carbon chains having at least one unsaturated bond such as a double bond in the alkylene group of 4 or more carbon chains.
- Examples thereof include an alkylene group having an unsaturated bond, and specific examples thereof include alkenylene groups such as a butenylene group, a pentenylene group, and a hexenylene group.
- the divalent aromatic group having 4 or more carbon chains may have at least 4 carbon chains, preferably 4 to 20 carbon atoms, more preferably 4 to 15 carbon chains, and 4 or more carbon chains.
- Examples thereof include divalent aromatic groups, and specific examples thereof include one (CH 2 ) p — o — C 6 H 4 — (CH 2 ) q — (p and q are each independently 0 or a natural number. And p + q ⁇ 2.).
- divalent substituted hydrocarbon group having 4 or more carbon chains at least one hydrogen atom in the divalent hydrocarbon group having 4 or more carbon chains is replaced with the above-mentioned substituent other than the hydroxyl group. And divalent substituted hydrocarbon groups.
- examples of the ring include an aliphatic ring and an aromatic ring.
- examples of the aliphatic ring include a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, and a cyclohexene ring.
- examples of the aromatic ring include a benzene ring.
- hydroxy fatty acid examples include ricinoleic acid, 11-hydroxypalmitic acid, rescuelaric acid, 10-hydroxyoxastearic acid, and 10-hydroxypalmitic acid. Ton precursor hydroxy fatty acid and the like.
- hydroxy fatty acids ricinoleic acid, 11-hydroxypalmitic acid, rescuelaric acid, 10-hydroxystearic acid, and 10-hydroxypalmitic acid are preferable.
- the hydroxy fatty acid derivatives used in the present invention include the above-described hydroxy fatty acids. Acid derivatives are preferred. Preferred examples of such hydroxy fatty acid derivatives include hydroxy fatty acid alkyl esters and hydroxy fatty acid dalyceride.
- hydroxy fatty acid alkyl ester examples include, for example, those represented by the general formula (3)
- R 4 represents an alkyl group, and! ⁇ 1 to! ⁇ 3 are the same as described above.
- the alkyl group represented by R 4 may be linear or branched, for example, has 1 or more carbon atoms, preferably has 1 to 10 carbon atoms, more preferably has 1 to 6 carbon atoms, and still more preferably has 1 to 6 carbon atoms.
- Examples include an alkyl group having a number of 1 to 3. Specific examples thereof include a methyl group, an ethyl group, an n-propyl group, and a 2-propyl group.
- Suitable hydroxy fatty acid alkyl esters in the present invention include hydroxy fatty acid alkyl esters having 1 to 3 carbon atoms in the alkyl ester portion (e.g., methyl ester, ethyl ester, propyl ester and the like).
- hydroxy fatty acid glyceride examples include hydroxy fatty acid monoglyceride, hydroxy fatty acid diglyceride, and hydroxy fatty acid triglyceride. Specific examples thereof include, for example, a compound represented by the general formula (4)
- R 6 to R 8 are each independently a hydrogen atom or a general formula (6) (In the formula,! ⁇ 1 to! ⁇ 3 are the same as described above.) However, at least one of R 6 to R 8 is a group represented by the general formula (6). ].
- Examples of the hydroxy fatty acid dalyceride represented by the general formula (4) include those obtained by condensing glycerin and the hydroxy fatty acid represented by the general formula (2).
- Hydroxy fatty acid glycerides suitable in the present invention include, for example, castor oil and the like.
- the hydrolysis product of the hydroxy fatty acid derivative used in the present invention can be produced by hydrolyzing the hydroxy fatty acid derivative.
- it is produced by subjecting hydroxy fatty acid dalyceride or hydroxy fatty acid alkyl ester to hydrolysis treatment chemically or enzymatically, for example, by treatment with a hydrolase such as lipase, alkali treatment, high pressure steam treatment, etc. can do.
- a hydrolase such as lipase, alkali treatment, high pressure steam treatment, etc.
- Examples of the hydrolysis product of the hydroxy fatty acid derivative include, but are not limited to, a hydrolysis product of castor oil, a hydrolysis product of 11-hydroxypalmitate and the like.
- the hydroxy fatty acid and hydroxy fatty acid derivative used in the present invention may be an R-form or S-form optically active form or a racemic form.
- the hydroxy fatty acid represented by the general formula (2) for example, the general formula (2-1)
- the obtained ratatone can be obtained as an optically active ratatone.
- R 1 and R 2 are the same group, the carbon atom to which R 1 and R 2 are bonded does not become an asymmetric carbon.
- the resulting ratatone can be obtained as an optically active ratatone.
- R 9 to R 11 are each independently a hydrogen atom or a general formula (6-1)
- R 9 to R 11 is a group represented by the general formula (6-1).
- the optically active hydroxy fatty acid dalyceride represented by the formula [1] is used, the resulting lactone can be obtained as an optically active ratatone.
- optically active hydroxy fatty acid glyceride examples include an optically active hydroxy fatty acid monoglyceride, an optically active hydroxy fatty acid diglyceride and an optically active hydroxy fatty acid triglyceride.
- the obtained optically active lactone is R-lactone, and if it is S-form, the obtained optically active lactone is S-form. —Lactones.
- optically active hydroxy fatty acid represented by the above general formula (2-1)
- optically active form of the above-mentioned hydroxy fatty acids include the optically active forms of the hydroxy fatty acids.
- optically active ricinoleic acid, optically active 1-hydroxyl mitinic acid, optically active rescuelaric acid, optically active 1 Preferred are 0-hydroxystearic acid and optically active 10-hydroxypalmitic acid.
- optically active hydroxy fatty acid alkyl ester represented by the above general formula (3-1) include optically active 11-hydroxypalmitate, optically active ethyl ricinoleate, optically active ethyl rescuellarate, Optically active 10-hydroxyethyl stearate, and optically active 10-hydroxyethyl palmitate are listed.
- optically active hydroxy fatty acid glyceride represented by the general formula (411) include, for example, optically active castor oil and the like.
- ⁇ -decalactone In order to produce y-decalactone as ratatones according to the production method of the present invention, it is not necessary to limit the production of hydroxy fatty acids, hydroxy fatty acid derivatives or hydrolyzation products of hydroxy fatty acid derivatives to castor, It is preferred to use at least one selected from the group consisting of oils, hydrolysates of castor oil, ricinoleic acid and rescuelaric acid. Further, for the production of ⁇ -decalactone by the production method of the present invention, it is possible to use, but not limited to, hydroxy fatty acids, hydroxy fatty acid derivatives or hydrolysis products of hydroxy fatty acid derivatives. It is preferable to use lumitic acid and phenyl or 11-hydroxyethyl palmitate.
- the hydroxy fatty acid, the hydroxy fatty acid derivative or the hydrolysis product of the hydroxy fatty acid derivative used in the present invention may be a commercially available product, an extract from a natural product, or an appropriately produced product. May be.
- hydrolysis products of hydroxy fatty acids, hydroxy fatty acid derivatives or hydroxy fatty acid derivatives may be obtained by a method other than chemical synthesis means. It is preferable to use the one obtained.
- 11-hydroxypalmitic acid that can be used for producing natural ⁇ -decalactone, a substance extracted from Garappa sasimaimo can be suitably used.
- optically active ratatones in the production method of the present invention, it is preferable to use an optically active substance as the xy fatty acid, the hydroxy fatty acid derivative or the hydrolysis product of the hydroxy fatty acid derivative.
- the medium for producing lactones comprises at least one selected from the group consisting of the above-mentioned hydroxyfatty acids, hydroxyfatty acid derivatives and hydrolysis products of hydroxyfatty acid derivatives. It is contained at a concentration of 10 to 50% (w Zv), preferably 15 to 25% (w / v) per 1 L.
- the medium for producing lactones is at least one cell selected from the group consisting of the above-mentioned hydroxyfatty acids, hydroxyfatty acid derivatives and hydrolysis products of hydroxyfatty acid derivatives. It can be prepared by appropriately adding other components (for example, nitrogen source and the like) usually used for culture. Other components to be added to the medium include, but are not limited to, yeast sources, urea, corn steep liquor, nitrogen sources including ammonium sulfate, diammonium hydrogen phosphate, etc., malt extract, polypeptone, glucose, etc.
- Additional carbon sources including sugars, manganese sulfate, calcium chloride, ferric chloride, ferrous sulfate, ferric sulfate, zinc sulfate, copper sulfate, magnesium sulfate, cobalt chloride, sodium molybdate, boron and boron
- Inorganic salts such as potassium chloride, flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD), nicotinamide adodenine dinucleotide (NAD), nicotinamide adodenine dinucleotide (NAD P) and konzaim A (CoA) and other coenzymes; nucleotides such as adenosine triphosphate (ATP); Vitamins such as Nichin, and sterile water, and the like.
- FMN flavin mononucleotide
- FAD flavin adenine dinucleotide
- NAD
- Cofactors such as inorganic salts, coenzymes, and vitamins can further increase the production of ratatone precursor hydroxy fatty acids and amino acids or lactones, but usually only a small amount is required. .
- One skilled in the art can readily determine, as needed, such other components to add to prepare a medium in which Candida sorbophila can grow.
- Candida 'Solpofila (Candida) described in (1) above is used.
- Candida sorbophila allows the ratatotone precursor hydroxy fatty acid and Z or lactones to be cultured in the medium. It can be produced inside.
- Ratatone precursor hydroxy fatty acids produced by Candida sorbophila can be converted to lactones by lactonization.
- Ratatonation of the lactone precursor hydroxy fatty acid can be performed by any method known to those skilled in the art, for example, by heat treatment under acidic conditions. Specifically, for example, by heating a culture containing the lactone precursor hydroxy fatty acid at 100 ° C. for 20 minutes under conditions of 13 to 5, the ratatone precursor in the culture is heated. It can ratatize hydroxy fatty acids.
- the compound in which the lactone precursor hydroxy fatty acid is ratatized is a lactone according to the present invention. In the present invention, it is not necessary to use a pH adjuster, and ratification can be achieved.
- Lactones produced by ratation of lactone precursor hydroxy fatty acids or ratatones produced by Candida sorbophila can be prepared by methods known to those skilled in the art. And Z or isolation and purification.
- lactone precursor hydroxy fatty acid means a hydroxy fatty acid that can be lactonized.
- the ⁇ lactone precursor hydroxy fatty acid '' in the present invention preferably has 4 or more carbon atoms having a hydroxyl group at the 4- or 5-position counted from the carbon atom of the carboxyl group, preferably has 5 to 22 carbon atoms, and preferably has 5 to 22 carbon atoms. Hydroxy fatty acids having 5 to 17 carbon atoms, more preferably 5 to 12 carbon atoms.
- Specific examples of the lactone precursor hydroxy fatty acid include, for example, the general formula (5)
- R 5 represents a divalent hydrocarbon group which may carbon chain optionally having a substituent is 2 or 3, R 1 and R 2 identical. Note that as above, R 2 and R 5 Bond to form a ring Good. ).
- R 5 is a divalent hydrocarbon group having 2 or 3 carbon atoms which may have a substituent; a divalent hydrocarbon group having 2 or 3 carbon atoms; and 2 or 3 having 2 or 3 carbon atoms. Valent substituted hydrocarbon groups.
- Examples of the divalent hydrocarbon group having a carbon chain of 2 or 3 include an alkylene group having an ethylene group or a trimethylene group having a carbon chain of 2 or 3, and an alkenylene having a carbon chain of an ethenylene group or a propenylene group of 2 or 3. And the like.
- the divalent substituted hydrocarbon group having a carbon chain of 2 or 3 at least one hydrogen atom of the divalent hydrocarbon group having a carbon chain of 2 or 3 is substituted with the above-mentioned substituents other than a hydroxyl group.
- Groups. Specific examples of the divalent substituted hydrocarbon group having 2 or 3 carbon chains include a propylene group.
- examples of the ring include an aliphatic ring and an aromatic ring.
- the aliphatic ring include a cyclobutane ring, a cyclopentane ring, a cyclohexane ring, and a cyclohexene ring.
- the aromatic ring include a benzene ring.
- lactone precursor hydroxy fatty acid in which R 5 in the general formula (5) is a hydrocarbon group of carbon atom 3 include ⁇ -hydroxydecanoic acid, and a hydroxy fatty acid represented by the following formula: And the like. 8Z
- Ratatones obtained by the production method of the present invention include, for example, those represented by the general formula (1) )
- ring A represents a ratatone ring
- R 1 and R 2 are the same as described above, and ring A and R 2 may combine to form a ring.
- examples of the lactone ring represented by ring A include a y-lactone ring represented by the following formula (7) or a ⁇ -lactone ring represented by the following (8). Further, the ring may have a substituent.
- ring A and R 2 combine to form a ring
- ring A and R 2 include, for example, a ring represented by the following formula.
- ratatones obtained by the production method of the present invention include, but are not limited to, lactones having 4 or more carbon atoms.
- R represents a protecting group
- the obtained lactone is an optically active lactone.
- optically active ratatones include, for example, a compound represented by the general formula (1-1)
- optically active lactone means a lactone that is an optically active substance (for example, R-form or S-form).
- the optically active ratatone produced by the method of the present invention produced by lactonization of hydroxy fatty acid, a lactone precursor produced from hydroxy fatty acid by Candida sorbophi la? , Or produced from hydroxy fatty acids by Candi da sorbophi la.
- optically active lactones obtained by the production method of the present invention include, but are not limited to, optically active lactones having 5 or more carbon atoms, preferably 5 to 12 carbon atoms.
- optically active lactones include lactones, and are preferably optically active ⁇ -decalactone, optically active ⁇ -valerolatonone, optically active two-hexalataton, and optically active y-heptalactone.
- optically active lactone of the present invention will be described in more detail by taking the production of optically active ⁇ -decalactone as an example.
- the production of the optically active lactone of the present invention can be carried out in principle by the same experimental operation as described in the method for producing this optically active ⁇ -decalactone.
- Optical activity medium used for culture (main culture) to produce hydroxydecanoic acid and Z or optically active dedecalactone.
- castor oil In the culture for producing optically active ⁇ -hydroxydecanoic acid and / or optically active ⁇ -decalactone in the present invention (corresponding to the main culture in the examples), castor oil, a hydrolysis product of castor oil, , At least one selected from the group consisting of ricinoleic acid and rescuelaric acid is used as a carbon source of the medium.
- the hydrolysis product of castor oil means a mixture obtained by hydrolyzing castor oil chemically or enzymatically.
- examples of the hydrolysis product of castor oil include a hydrolyzate obtained by hydrolyzing castor oil with lipase (hereinafter, referred to as “hydrolysis product by lipase”).
- the main component of the hydrolyzate obtained by hydrolyzing castor oil with lipase is ricinoleic acid. Therefore, the hydrolysis product of castor oil contains, for example, ricinoleic acid, which is a major constituent fatty acid of castor oil, as a main component.
- the lipase used for the hydrolysis of castor oil can be used without any particular limitation as long as it produces ricinoleic acid.
- Examples of the lipase include: (Meito Sangyo Co., Ltd.): Lipase OF, Lipase MY, (Amano Enzym Co., Ltd.): Newase F3G, Lipase A “Amano” 6, Lipase AY “Amano” 30G, Lipa F-AP15, lipase G "Amano” 50, lipase M “Amano” 10 and lipase R "Amano" G.
- the hydrolysis by lipase is carried out under the conditions for producing a hydrolysis product containing ricinoleic acid as a main component from castor oil.For example, 0.5 g of lipase is added to 100 g of castor oil at 30 ° C. 24 hours Obtained by encapsulation.
- the hydrolyzate of the lipase thus obtained can be used as it is in the form of a mixture.
- the hydrolysis product of castor oil is contained in the main culture medium of the present invention
- the hydrolyzate by lipase will be contained in the medium after the culture. This is because in the production method of the present invention, a hydrolysis product is produced from castor oil in the medium by Candida sorbophila used in the present invention. Therefore, if castor oil is added to the medium, adding a hydrolyzate by lipase is an optional step.
- At least one selected from the group consisting of castor oil, a castor oil hydrolyzate, ricinoleic acid and rescuelaric acid may be used.
- rescuelaric acid may be used alone or in an appropriate combination of two or more.
- these castor oil hydrolyzed products of castor oil, or ricinoleic acid or rescueraric acid, castor oil and / or hydrolyzed products of castor oil are preferred.
- the concentration at which at least one selected from the group consisting of castor oil, a hydrolyzate of castor oil, ricinoleic acid and rescuelaric acid is added to the culture medium is 1 L of culture medium.
- it is about 10 to 50% (w / v), preferably about 15 to 25% (w / v).
- the medium used in the present invention may further contain components such as yeast extract, malt extract, polybeptone, and glucose, and the medium containing such additional components may be used as a nutrient medium.
- gamma - it can be used for producing optically active ⁇ - Dekaraku tons arsenide Dorokishidekan acid and ⁇ or invention.
- yeast extract urea, corn steep liquor, ammonium sulfate, diammonium hydrogen phosphate, and the like can be used by being contained in the medium as a nitrogen source.
- sugars such as malt extract, polypeptone, and glucose can be used by being added to the medium as an additional carbon source.
- a culture medium castor oil, a hydrolysis product of castor oil, ricinoleic acid or A synthetic medium containing at least one selected from the group consisting of rescuelaric acid as a sole carbon source may be used.
- additional components such as the above-mentioned additional nitrogen source or carbon source may be further added to this synthetic medium.
- optically active y-hydroxydecanoic acid and Z or optically active ⁇ -decalactone it is possible to further increase the production of optically active y-hydroxydecanoic acid and Z or optically active ⁇ -decalactone.
- cofactors include manganese sulfate, calcium chloride, ferric chloride, ferrous sulfate, ferric sulfate, zinc sulfate, copper sulfate, magnesium sulfate, cobalt chloride, sodium molybdate, boron, and potassium iodide.
- Inorganic salts such as flavin mononucleotide (FMN), flavin adenine dinucleotide (FAD), nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate (NAD P), coenzyme A (CoA), etc.
- Co-enzymes nucleotides such as adenosine triphosphate (ATP), and vitamins such as L-carnitine. The addition amount of the cofactor may be very small.
- the main culture medium used in the present invention is selected from, for example, a group consisting of castor oil, a hydrolysis product of castor oil, ricinoleic acid, or resqueuric acid in a YM medium, a potato dextrose medium, a potato sucrose medium, or the like.
- the medium include a medium to which at least one is added, or a medium to which the above additional components are arbitrarily added.
- Candida sorbophila according to the present invention is cultured in the above-mentioned medium a) to produce and accumulate ⁇ -hydroxydecanoic acid and / or optically active y-decalactone in the medium. be able to.
- the Candida sorbophila according to the present invention may be directly inoculated into the medium of the above a) and cultured, but it is preferable that the Candida sorbophila previously seed-cultured in a normal medium is used as the seed culture. Inoculate or inoculate the medium of a) and culture.
- the culture conditions for this seed culture are ⁇ -hydroxydecanoic acid and Z or optically active ⁇ -de
- the culture may be the same as the culture for the production of caractone (main culture), but is not particularly limited as long as the Candida sorbophila strain to be used can grow.
- the usual medium used for seed culture may be a solid medium or a liquid medium, and contains at least one selected from the group consisting of castor oil, a hydrolyzate of castor oil, ricinoleic acid, and rescueraric acid. It is not necessary to do so, but it may be contained.
- a YM slant agar medium or a potato dextrose slant agar medium can be mentioned.
- the amount of the seed culture added to the medium for the main culture is not particularly limited. However, for example, when the seed culture is a liquid culture, it is preferable to add a culture having an absorbance of 0D610 at a concentration of about 30 to 1 to 3% of the medium during the main culture.
- the preculture may be performed, and then the main culture in the medium a) may be performed.
- the culture conditions for the preculture may be the same as those for the main culture, but are not particularly limited as long as the Candida sorbophila strain to be used can grow.
- the medium used for seed culture does not need to contain at least one selected from the group consisting of castor oil, a hydrolyzate of castor oil, ricinoleic acid, and rescuellaric acid, but does not. You may.
- the culture conditions used are aerobic conditions.
- the culturing temperature is appropriately selected usually from the range of 20 to 35 ° C, preferably 24 to 30 ° C.
- the pH of the medium is appropriately selected usually from the range of 5 to 7, preferably from 5.5 to 6.5.
- the culture is performed, for example, in a shake flask or in a fermenter (eg, a fermenter equipped with a stirring and aeration device).
- the culture time of the main culture is not particularly limited as long as it is a time sufficient to produce optically active ⁇ -hydroxydecanoic acid and ⁇ or optically active ⁇ -decalactone, but preferably, optically active ⁇ -hydroxydecanoic acid and // Select the time to reach the maximum optically active ⁇ -decalactone production.
- the time at which the production of the optically active ⁇ -hydroxydecanoic acid and / or optically active ⁇ -decalactone reaches the maximum depends on the medium composition, castor oil to be added as a base, hydrolysis products of castor oil, ricinoleic acid or resquerae At least one addition amount selected from the group consisting of acids, depending on the culture equipment used Ventilation according to the value.
- the culturing time may be appropriately selected from the range of usually 1 hour to 30 days, preferably 12 hours to 25 days, for example, for culturing using a shake flask as a culturing device.
- this culture using a fermenter it may be appropriately selected from a range of usually 1 day to 20 days, preferably 3 days to 10 days.
- the use of a fermentation tank is preferable from the viewpoint of production efficiency because culturing in a relatively short time is sufficient.
- the above culture time is determined by sampling the optically active ⁇ -decalactone produced in the medium over time, or sampling the ⁇ -hydroxydecanoic acid produced in the medium over time, Measure the amount of optically active ⁇ -zap rataton produced by tonji-dori by gas chromatography (GC), thin-layer chromatography (TLC), or gas chromatography / mass spectrometry (GC-MS).
- GC gas chromatography
- TLC thin-layer chromatography
- GC-MS gas chromatography / mass spectrometry
- an optically active one from optically active ⁇ over arsenate Dorokishidekan acid gamma - Dekaraku ton thus prepared optically active ⁇ - arsenate Dorokishidekan acid in the medium is produced; if Ru is used as it is as a pharmaceutical intermediates, etc.
- the ⁇ / -hydroxydecanoic acid is further converted to optically active decalactone by lactone and used to produce the optically active ⁇ -decalactone.
- the ratatonization may be performed by any known ratatonization method.
- the ratatonization of the optically active ⁇ -hydroxydecanoic acid of the present invention is carried out using the y produced in the above-mentioned medium.
- Hydroxydecanoic acid may be recovered by a conventional method, and the culture may be performed directly after ratification, and the culture medium may contain optically active ⁇ -hydroxydecanoic acid. It may be performed in a state.
- lactonization in a culture solution conventionally performed can be used.
- Specific examples thereof include a method in which an acid such as dilute hydrochloric acid or dilute sulfuric acid is added to a culture solution after completion of the culture to acidify ⁇ ⁇ of the culture solution.
- ratatonization is performed without acidifying ⁇ of the culture solution. It is preferable to use the method performed in the state described above.
- the pH of the culture solution at the end of the culture is already inclined toward the acidic side in the range of 3.0 to 4.5, so that the pH of the culture solution is not acidified, Can be obtained.
- the culture under the acidic conditions is contained in the culture by heating for about 10 minutes to 1 hour, preferably for about 10 to 30 minutes, without performing the treatment for acidifying pH.
- the lactonization of optically active ⁇ -hydroxydecanoic acid can be achieved.
- the heating temperature is set to a temperature range of about 70 to 130 ° C, preferably about 90 to 120 ° C.
- the medium may be under acidic conditions, and the pH is preferably 2 to 5.
- the optically active y-hydroxydecanoic acid can be converted to optically active ⁇ -decalactone by lactonization of such optically active ⁇ -hydroxydecanoic acid.
- optically active 0-decalactone is separated and removed from the culture by a method such as centrifugation, and then recovered and purified from the culture by a conventional method such as solvent extraction and distillation.
- R- ⁇ -decalactone can be obtained with high optical purity.
- the lactones obtained by the production method of the present invention can be used for flavors, pharmaceutical intermediates and the like.
- R- ⁇ -decalactone can be used to add or enhance the functionality of beverages, chewing gum, fruit juice, tobacco products, pharmaceutical preparations, flavors, flavored products, etc.
- Ton has the advantage of a stronger aroma and a more natural fruity character as compared to S- ⁇ -decalactone [(A. Mosandle et al., J. Agric. Food Chem,. 37, 413 (1989 )]
- the production method of the present invention is characterized in that Candida sorbophila is used.
- optically active ratatones such as R- ⁇ -decalactone and R- ⁇ _decalactone can be obtained with high optical purity.
- the production efficiency of Ry-decalactone is high.
- the high production efficiency of R- ⁇ -decalactone that can be achieved by the production method of the present invention is probably due to the fact that C- Candida sorbophila was obtained in the production system of the present invention.
- the FC58 strain was inoculated on a YM slant agar medium and activated by culturing at 27 ° C. for 3 days. Also, as a medium for pre-culture, add 0.09 ⁇ yeast extract, 0.09 malt dextrin, 0.15 g of polyptone and 0.3 g of glucose to a 30 OmL Erlenmeyer flask, and add distilled water to distilled water. Was adjusted to a total volume of 30 mL, pH 6, and sterilized with an auto turret at 121 ° C for 15 minutes. The cooled FC58 medium was inoculated with the activated FC58 strain as described above, and was subjected to 150 rp at 27 ° C using a rotary shaking culture apparatus.
- the medium after shaking culture at m for 24 hours was used as a pre-culture solution. Further, a main culture medium for the subsequent main culture was prepared. That is, 0.3 g of yeast extract, 0.3 g of malt extract and 0.5 g of polypeptone are added to a 50 OmL volumetric flask, and distilled water is added to make a total volume of 10 OmL, pH 6, and 20 g of castor oil is further added. The added product was prepared by autoclaving at 121 ° C for 15 minutes. Next, 2 mL of the above-mentioned preculture solution was inoculated into the cooled main culture medium, and the main culture was performed by shaking culture at 150 rpm at 27 ° C. No pH adjustment was performed during the culture.
- the pH of the culture solution after the culture was 4.06. From the third day after the start of the culture, 5 mL of the culture solution is aseptically sampled over time, and each of the sampled culture solutions (samples) is heated at 100 ° C for 20 minutes to obtain the lactone of ⁇ -hydroxydecanoic acid. Was performed. After such ratatonization, this sample was extracted with ethyl acetate, and the separated organic layer was quantified by gas chromatography (GC) analysis using the internal method (using ethyl decanoate as an internal standard sample). As a result, the amount of R- ⁇ -zachalactone produced in the culture solution on day 14 after the start of the cultivation was maximized. The production amount of R-y-decalactone obtained from the culture solution 14 days after the start of the culture was 8.41 g / L of the main culture medium. The optical purity was at least 99% ee.
- Example 2 The optical purity was at least 99% ee.
- R- ⁇ -decalactone was produced in the same manner as in Example 1 except that 20 g of castor oil was replaced by 2 Og of ricinoleic acid (purity: 80% or more; manufactured by Wako Pure Chemical Industries, Ltd.). Was. As a result, the amount of R- ⁇ -decalactone produced in the culture solution on day 20 after the start of the culture was maximized.
- the production amount of Ry-decalactone obtained from the culture solution 20 days after the start of the culture was 21.89 g / L of the main culture medium. The optical purity was 99% e e or more.
- the present culture medium manganese sulfate (Mn S0 4 'H 2 0 ) 1. 7m g, calcium chloride (C a C 1 2 ⁇ H 2 0) 0. 5 5 mg, ferric chloride (F e C 1 3 ⁇ H 2 0) 0.3 7 5 mg, zinc sulfate (Z n S 0 4 'H 2 0) 2. 2m g, sulfate ⁇ (C u S 0 4' H 2 0) 0. 4mg, magnesium sulfate (Mg S_ ⁇ 4 'H 2 ⁇ ) 5 . 9 mg, chloride Kobanore Doo (C o C 1 2 ⁇ H 2 ⁇ ) 0.
- the FC58 strain was inoculated on a YM slant agar medium and activated by culturing at 27 ° C. for 3 days.
- a medium for pre-culture add 0.3 g of yeast extract, 0.3 g of malt extract, 0.5 g of polypeptone and 1.O g of glucose to a 50-OmL volumetric flask, add distilled water and add 1 g.
- a solution having a pH of 00 ml and a pH of 6 was prepared by sterilizing in an autoclave at 121 ° C. for 15 minutes.
- the cooled culture medium was inoculated with the activated FC58 strain as described above, and subjected to shaking culture at 150 rpm for 24 hours at 27 ° C using a rotary shaking culture apparatus. Liquid. Further, a main culture medium for the subsequent main culture was prepared. That, 5 L volume changer over Sulfur yeast Mentor E key scan 6. O g, malt extract 6 ⁇ 0 g, polypeptone 1 0. 0 g, manganese sulfate (M n S 0 4 ⁇ H 2 0) 34 mg, chloride calcium (C a C 1, - H 2 0) 1 1 mg, ferric (F e C 1 3 ⁇ H 2 0) salt of 7.
- Ricinoleic acid (purity 80% or more; manufactured by Wako Pure Chemical Industries, Ltd.) Hydrolyzate of castor oil 600 g instead of 400 g with lipase (Lipase OF; manufactured by Meito Sangyo Co., Ltd.)
- R- ⁇ -decalactone was produced in the same manner as in Example 4 except for using.
- the amount of R- ⁇ -decalactone produced in the culture solution 5 days after the start of the culture was maximized.
- the production amount of R- ⁇ -decalactone obtained from the culture solution 5 days after the start of the culture was 40.50 g / L of the main culture medium.
- the optical purity was more than 99% e e.
- Candida sorbophila ATCC 74362 strain In place of Candida sorbophi la FC58 strain, Candida sorbophila ATCC 74362 strain, Candida sorbophila ATCC60130 strain or Candida sorbophila other than Candida sorbophila strain AT015 using Candida sorbophila An optically active ⁇ -decalactone was produced in the same manner as in Example 2. As a result, when each strain was used, the amount of ⁇ -decalactone produced was maximum for the culture solution on day 19, day 11, and day 11 after the start of culture, respectively. . From the culture that produced such a maximum amount The production amounts of the obtained ⁇ -decalactone were 13.75 g, 12.97 g, and 12.9.7 g per 1 L of the main culture medium, respectively.
- R- ⁇ -decalactone was produced in the same manner as in Example 1 except that the sampled culture solution (sample) was not subjected to ratatonization treatment by heating. As a result, the amount of R- ⁇ -decalactone produced in the culture solution on the 14th day after the start of the culture was the largest. The amount of R_ ⁇ -decalactone obtained from the culture solution on day 14 after the start of the culture was 8.41 g per 1 L of the main culture medium. The optical purity was more than 99% e e.
- R- ⁇ -decalactone was produced in the same manner as in Example 2, except that the sampled culture solution (sample) was not subjected to lactonization treatment by heating. As a result, the amount of R- ⁇ -decalactone produced in the culture solution on day 20 after the start of the culture was maximized. The production amount of R- ⁇ -decalactone obtained from the culture solution 20 days after the start of the culture was 21.89 g per 1 L of the main culture medium. Optical purity was greater than 99% e e. Example 9
- R- ⁇ -decalactone was produced in the same manner as in Example 3 except that the sampled culture solution (sample) was not subjected to lactonization treatment by heating. As a result, the amount of R- ⁇ -decalactone produced in the culture solution on day 20 after the start of the culture was maximized.
- the production amount of R- ⁇ -decalactone obtained from the culture solution 20 days after the start of the culture was 27.37 g / L of the main culture medium. Optical purity was greater than 99% e e.
- Example 11 Each sampled culture solution (sample) undergoes lactonization by heating. Except for the absence, R- ⁇ -decalactone was produced in the same manner as in Example 4. As a result, the amount of R_T-decalactone produced in the culture solution on day 10 after the start of the culture was the largest. The production amount of R- ⁇ -decalactone obtained from the culture solution on day 10 after the start of the culture was 49.94 g / L of the main culture medium. Optical purity was more than 99% ee.
- Example 11 Example 11
- RT-/-decalactone was produced in the same manner as in Example 5, except that the sampled culture solution (sample) was not subjected to lactonization treatment by heating. As a result, the amount of R- ⁇ -decalactone produced in the culture solution 5 days after the start of the culture was the largest. The production amount of R-y-decalactone obtained from the culture solution 5 days after the start of the culture was 40.50 g per 1 L of the main culture medium. The optical purity was more than 99% e e.
- Example 1 2 The optical purity was more than 99% e e.
- R- ⁇ -decalactone was produced in the same manner as in Example 6, except that the sampled culture solution (sample) was not subjected to lactonization treatment by heating. As a result, when each strain was used, the amount of R- ⁇ -decalactone produced in the culture solution on the 19th, 11th, and 11th days after the start of cultivation reached the maximum, respectively. Was. The production amount of R- ⁇ -decalactone obtained from the culture with the maximum production amount was 13.75 g, 12.97 g, and 12 97 g. Example 13
- the FC58 strain was inoculated on a YM slant agar medium, cultured at 27 for 3 days, and activated.
- a medium for pre-culture 0.09 g of yeast extract, 0.09 of malt extract, 0.158 of polypton and 0.3 g of glucose were placed in a 30-OmL Erlenmeyer flask, and distilled water was added.
- a total volume of 3 OmL and pH 6 was prepared by sterilizing with an autoclave at 121 ° C for 15 minutes.
- the cultured FC58 strain was inoculated and subjected to shaking culture at 27 ° C. and 150 rpm for 24 hours using a rotary shaking culture apparatus to obtain a pre-culture liquid.
- a main culture medium for subsequent main culture was prepared. That, 3 0 0 m L Yozaka necked flask yeast extract 0. 0 9 g, malt extract 0. 0 9 g and Poripepu tons 0. 1 5 g, manganese sulfate (Mn S 0 4 ⁇ H 2 0) 0. 5 1 mg, chloride calcium ⁇ beam (C a C 1 2 ⁇ ⁇ 2 ⁇ ) 0. 1 7 mg, ferric (F e C 1 3 ⁇ H 2 0) 0. 1 1 mg chloride, zinc sulfate (Z n S 0 4 ⁇ H 2 0 ) 0. 6 6 mg, copper sulfate (C u S 0 4 ⁇ H 2 0) 0.
- S- ⁇ -decalactone was produced in the same manner as in Example 13 except that the sampled culture solution (sample) was not subjected to ratatonization treatment by heating. As a result, the S- ⁇ -decalactate produced in the culture solution on day 11 after the start of culture The amount of the gas became the maximum. The amount of S- ⁇ -decalactone obtained from the culture solution on day 11 after the start of the culture was 0.019 g per 3 O ml of the main culture medium, and the optical purity was 96% ee. That was all.
- Reference example 1
- the ⁇ -decalactone production was performed in the same manner as in Example 2 except that the Yarrowia lipolytica IF00717 strain, which is generally known for its higher ⁇ -decalactone producing ability, was used. Manufactured. As a result, the amount of ⁇ -decalactone produced in the doubled solution on day 6 after the start of the culture was maximized. The production amount of ⁇ -decalactone obtained from the culture solution on day 6 after the start of the culture was 4.90 g per 1 L of the main culture medium.
- ratatones for example, optically active lactones such as optically active ⁇ -decalactone (such as R- ⁇ -decalactone) and optically active ⁇ -decalactone, are easily obtained with high production efficiency. be able to.
- the lactone precursor hydroxy fatty acid for example, ⁇ -hydroxydecanoic acid ⁇ -hydroxydecanoic acid can be efficiently produced in the intermediate step of the production method of the present invention.
- the production method of the present invention does not require the use of an emulsifier or a ⁇ adjuster in the medium, and provides a method for producing a hydroxy fatty acid, a hydroxy fatty acid derivative, and / or a hydrolysis product of a hydroxy fatty acid derivative as a carbon source. Since it can be used by adding it to a medium at a high concentration useful for large-scale production, it can be used very advantageously in industrial production.
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AU2003244011A AU2003244011A1 (en) | 2002-06-28 | 2003-06-27 | Processes for producing lactone |
US10/519,212 US7129067B2 (en) | 2002-06-28 | 2003-06-27 | Method for producing lactone |
EP03736294.4A EP1538215B1 (en) | 2002-06-28 | 2003-06-27 | Processes for producing lactone |
JP2004517318A JP4145297B2 (ja) | 2002-06-28 | 2003-06-27 | ラクトン類の製造方法 |
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Cited By (2)
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JP2008518595A (ja) * | 2004-11-03 | 2008-06-05 | ヴェ. マヌ フィル | γ−ラクトンの立体選択的合成 |
DE102012207921A1 (de) | 2012-05-11 | 2013-11-14 | Evonik Industries Ag | Mehrstufiges Syntheseverfahren mit Synthesegas |
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- 2003-06-27 AU AU2003244011A patent/AU2003244011A1/en not_active Abandoned
- 2003-06-27 WO PCT/JP2003/008217 patent/WO2004003213A1/ja active Application Filing
- 2003-06-27 JP JP2004517318A patent/JP4145297B2/ja not_active Expired - Lifetime
- 2003-06-27 EP EP03736294.4A patent/EP1538215B1/en not_active Expired - Lifetime
- 2003-06-27 US US10/519,212 patent/US7129067B2/en not_active Expired - Lifetime
- 2003-06-27 CN CNB038196417A patent/CN100473727C/zh not_active Expired - Lifetime
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JPS6066991A (ja) * | 1983-09-24 | 1985-04-17 | Kanebo Ltd | 品質の改良されたヒマシ油の製造法 |
JPS61195693A (ja) * | 1985-02-23 | 1986-08-29 | Kanebo Ltd | 品質の改良されたヒマシ油の製造法 |
EP0258993A2 (en) * | 1986-07-28 | 1988-03-09 | Unilever Plc | Production of lactones |
EP0356291A1 (fr) * | 1988-08-04 | 1990-02-28 | Pernod-Ricard | Procédé de production microbiologique de décanolide gamma (R) et d'octanolide gamma (R) |
EP0371568A1 (en) * | 1988-12-01 | 1990-06-06 | Unilever N.V. | Process for producing gamma-lactones |
JPH04108782A (ja) * | 1990-08-30 | 1992-04-09 | Takasago Internatl Corp | 光学活性γ―ブチロラクトン誘導体の製造方法 |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2008518595A (ja) * | 2004-11-03 | 2008-06-05 | ヴェ. マヌ フィル | γ−ラクトンの立体選択的合成 |
DE102012207921A1 (de) | 2012-05-11 | 2013-11-14 | Evonik Industries Ag | Mehrstufiges Syntheseverfahren mit Synthesegas |
WO2013167663A2 (de) | 2012-05-11 | 2013-11-14 | Evonik Industries Ag | Mehrstufiges syntheseverfahren mit synthesegas |
Also Published As
Publication number | Publication date |
---|---|
JPWO2004003213A1 (ja) | 2005-10-27 |
CN100473727C (zh) | 2009-04-01 |
US7129067B2 (en) | 2006-10-31 |
EP1538215A1 (en) | 2005-06-08 |
AU2003244011A1 (en) | 2004-01-19 |
US20050130278A1 (en) | 2005-06-16 |
JP4145297B2 (ja) | 2008-09-03 |
EP1538215B1 (en) | 2014-11-12 |
EP1538215A4 (en) | 2010-09-22 |
CN1675370A (zh) | 2005-09-28 |
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