JP2001069968A - New microorganism ok4 strain and biological production of polyhydroxyalkanoate using the microorganism - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new microorganism which can simultaneously simply produce polyhydroxyalkanoates containing hydroxybutyric acid and so on as monomer units, respectively, as biodegradable polyesters in high yields. SOLUTION: This microorganism can simultaneously produce polyhydroxyalkanoates containing 3-hydroxybutyric acid and a 3-3- hydroxy-5-(4-substituted phenyl) valeric acid of formula I (R is H or CH3) as monomer units, respectively. The method for biologically producing the polyhydroxyalkanoates comprises preferably culturing the new microorganism: Burkholderia sp. OK4 strain (FERM P-17371) in a culture medium containing a 5-(4-substituted phenyl) valeric acid of formula II and then extracting the polyhydroxyalkanoates containing 3-hydroxybutyric acid and the 3-hydroxy-5-(4- substituted phenyl) valeric acid of formula I (R is common to that of the compound of formula II) as monomer units, respectively, from the culture product.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel microorganism capable of producing a biodegradable polyester, and a method for producing a biodegradable polyester biologically using the microorganism. Specifically, as a biodegradable polyester, a novel microorganism that simultaneously produces and accumulates 3-hydroxybutyric acid and a polyhydroxyalkanoate containing 3-hydroxy-5- (4-substituted phenyl) valeric acids as monomer units, And a method for simultaneously producing a polyhydroxyalkanoate containing 3-hydroxybutyric acid and 3-hydroxy-5- (4-substituted phenyl) valeric acids as monomer units using the microorganism.

[0002]

2. Description of the Related Art For many years, synthetic polymers derived from petroleum have been used as plastics and the like, but treatment of these plastics and the like contained in household waste has become a major social problem. In the past, synthetic polymer materials derived from petroleum have been used as substitutes for metal materials, glass, etc. due to the advantage that they are not easily decomposed, but in recent years they are consumed in large quantities and are discarded in large quantities. Was devastated and accumulated at waste disposal sites. In addition, when incineration is performed, the amount of CO ₂ emission increases, and in some cases, it also causes the generation of harmful substances such as dioxins and environmental hormones. In consideration of such environmental effects, instead of conventional synthetic polymer materials derived from petroleum,
While maintaining a certain strength and durability, the use of biodegradable polymer materials that can be decomposed by various naturally occurring microorganisms is being studied, and some of them are being put to practical use.

For example, a microorganism-produced polyester (polyhydroxyalkanoate; PHA) represented by poly-3-hydroxybutyrate (PHB) has a property of being degradable by living organisms, and has a high biodegradability. Application as a molecular material is being studied. That is, since the microorganism-produced polyester is biodegraded again by microorganisms or the like, it is finally taken into natural material circulation. That is, it can be used as a plastic that enables environmental conservation. In addition, there is a possibility that the medical soft member will be more useful in the future (JP-A-5-159, JP-A-6-169980, JP-A-6-169988, JP-A-6-169988). -225921
No., etc.).

[0004] It has been reported that many bacteria produce and accumulate poly-3-hydroxybutyrate (PHB) or a copolymer of 3-hydroxybutyrate and other hydroxyalkanoates in the cells. (“Biodegradable Plastic Handbook” (edited by Biodegradable Plastics Study Group; NTT Co., Ltd.), p.178
-197). For example, Alcaligenes eutropus H16 strain (ATCC N
o. 17699) and variants thereof have been studied in detail for the production of these polymers. That is, by changing the carbon source serving as a substrate, a copolymer of 3-hydroxybutyrate (3HB: 3-hydroxybutanoic acid) and 3-hydroxyvalerate (3HV: 3-hydroxypentanoic acid) or each of both is obtained. It has been published that various ratios of copolymers having both units as components are produced (Japanese Patent Application Laid-Open Nos. 6-15604 and 7-1435).
No. 2, JP-B-8-19227, etc.).

[0005] Also, in Japanese Patent Publication No. 2642937,
Pseudomonas oleovo
rans) By giving acyclic aliphatic hydrocarbons as a carbon source to ATCC 29347 strains,
It is disclosed to produce a polyester having -hydroxyalkanoate (3HA: 3-hydroxyalkanoic acid) as a monomer unit.

[0006] Japanese Patent Application Laid-Open No. Hei 5-74492 discloses a genus Methylobacterium, Paracoccus (Pa).
by contacting bacteria of the genus racoccus, genus Alcaligenes, and genus Pseudomonas with a primary alcohol having 3 to 7 carbon atoms,
A method for producing a copolyester of 3HB and 3HV is disclosed.

[0007] JP-A-5-93049 and JP-A-7-265065 disclose Aeromonas caviae with unsaturated fatty acids such as oleic acid ((Z) -9-octadecenoic acid) and oleic acid. Cultivation using olive oil containing a large amount of unsaturated fatty acids such as 3HB and 3-hydroxyhexanoate (3
(HHx: 3-hydroxyhexanoic acid) is disclosed to produce a two-component copolymerized polyester.

Japanese Patent Application Laid-Open No. 9-191893 discloses Comamonasacidovorans IFO 1
By culturing strain 3852 in a medium containing D-gluconic acid and 1,4-butanediol as a carbon source, a polyester having 3HB and 4-hydroxybutyrate (4HB: 4-hydroxybutanoic acid) as a monomer unit Is disclosed.

Further, various substituents have been introduced to impart various functions to polyesters produced by these microorganisms. Regarding the introduction of these substituents, for example, FEMS Microbiology Letters, 128 (1995)
It is described in detail on pages 219-228 and the like. Examples of the substituent (functional group) to be introduced include an unsaturated hydrocarbon group, an ester group (such as a lactone structure), an aryl group (aromatic ring),
Examples include a cyano group, a halogenated hydrocarbon group, and an epoxy group (epoxidation).

As described above, a polyhydroxyalkanoate (PHA), which is a microorganism-produced polyester, has been shown to exhibit various functions by introducing various substituents into a monomer unit. In terms of further expansion, P containing a wider range of monomer units
A method for producing HA is desired. In addition, it is necessary to search for and develop a microorganism capable of producing the PHA.

For example, as a microorganism producing PHA containing an aryl group in a monomer unit molecule, 3-hydroxy-5-phenylvaleric acid (3H
One strain of Pseudomonas oleovorans is reported as a strain that introduces PV: 3-hydroxy-5-phenylpentanoic acid, a compound of the above formula (1), wherein R = H) as a monomer unit into PHA. (Macromolecules, 24, p. 5256-526)
0 (1991)). The above report, and International J
ournal of Biological Macromolecules, 19, p.29-34
Referring to (1996), the above strain uses nonanoic acid or octanoic acid as a growth substrate, and is transformed from 5-phenylvaleric acid (PVA: 5-phenylpentanoic acid) added to the medium.
Produce PHA containing HPV as a monomer unit. The other PHA monomer units simultaneously produced at this time include 3-hydroxyvaleric acid (3HV: 3-hydroxypentanoic acid) and 3-hydroxyhexanoic acid (3H
Hx), 3-hydroxyheptanoic acid (3HHp), 3-hydroxyoctanoic acid (3HO), 3-hydroxynonanoic acid
(3HN), 3-hydroxydecanoic acid (3HD), and 3-hydroxydodecanoic acid (3HDO), and contains a 3-hydroxybutyric acid (3HB: 3-hydroxybutanoic acid) unit which is a main component of general PHA. Not. In addition, Mac
In romolecules, 29, p.1762-1766 (1996), 5- (p
-Tolyl) valeric acid (5- (p-tolyl) pentanoic acid), 5- (4-ethylphenyl) valeric acid (5-
Using (4-ethylphenyl) pentanoic acid), 5- (4-biphenylyl) valeric acid (5- (4-biphenylyl) pentanoic acid) and 8- (p-tolyl) octanoic acid as substrates, corresponding to the respective substrate compounds Containing aromatic ring
It is described to produce HA. Also, Macromole
cules, 29, p.3432-3435 (1996), a PHA containing an aromatic ring corresponding to each substrate compound using 6-phenoxyhexanoic acid, 8-phenoxyoctanoic acid, and 11-phenoxyundecanoic acid as substrates. Production is described. However, 3HB in the monomer unit
Is not included.

[0012]

On the other hand, in order to meet various needs, PHA having various physical properties is desired. For example, as a substituent, 3HB and an aryl group such as a phenyl group, which have not been reported so far, are used. There is a strong demand for a method for producing a copolymer containing simultaneously 3-hydroxyalkanoic acid (3HA). In addition, great expectations are placed on the search and development of microorganisms that produce the copolymer.

The present invention has been made to solve the above problems, and an object of the present invention is to provide 3HB and 3-hydroxy-5- (4-substituted phenyl) valeric acid represented by the following general formula (1). An object of the present invention is to provide a novel microorganism which simultaneously produces polyhydroxyalkanoate containing a monomer unit. Another object of the present invention is to provide a polyhydroxyalkanol containing 3HB and 3-hydroxy-5- (4-substituted phenyl) valeric acid represented by the following general formula (1) as monomer units by utilizing the above-mentioned microorganism. It is an object of the present invention to provide a biological production method of an ate.

[0014]

Means for Solving the Problems In order to solve the above-mentioned problems, the present inventors isolated a large number of strains from soils newly collected from various places, and obtained 5-phenylvaleric acid (PVA:
PVA-assimilating and growing microorganisms growing on a selective medium using 5-phenylpentanoic acid) as the sole carbon source were selected. Furthermore, the genus was identified and the production ability of polyhydroxyalkanoate (PHA) was examined for a plurality of the selected strains of PVA-assimilating microorganisms, and nonanoic acid and 5-phenylvaleric acid were used as substrates (carbon source). ), When 3-hydroxybutanoic acid (3HB) and 3
-Hydroxy-5-phenylvaleric acid (3HPV
A strain showing the ability to simultaneously produce PHA containing A) as a monomer unit was found. Among them, the strain having the above-mentioned PHA-producing ability and capable of being identified as Burkholderia sp. (Burkholderia sp.) Was determined to be a novel strain, and one of them was identified as Burkholderia sp. ) As OK4 strain,
Deposited with the Institute of Biotechnology, National Institute of Advanced Industrial Science and Technology (NIBH) (Deposit No .: FERM P-17371). Further, when this OK4 strain was further cultured in a medium containing 5- (4-substituted phenyl) valeric acid represented by the following general formula (2), 3-hydroxybutanoic acid (3HB) and the corresponding 3-hydroxy- 5- (4-substituted phenyl)
It has been found that PHA containing valeric acid as a monomer unit can be produced. Based on such knowledge, the present invention has been completed.

That is, the microorganism capable of producing polyhydroxyalkanoate of the present invention comprises 3-hydroxybutyric acid and the following general formula (1):

[0016]

Embedded image (Wherein R represents H or CH ₃ ) A novel microorganism Burkholderia species simultaneously producing polyhydroxyalkanoate containing 3-hydroxy-5- (4-substituted phenyl) valeric acid as a monomer unit (Burkholderia sp.) OK4 strain (FERM P-17371).

Further, the method for biologically producing polyhydroxyalkanoate of the present invention is characterized in that the Burkholderia sp. OK4 strain (FERM P-17371) is used.
With the following general formula (2):

[0018]

Embedded image (Wherein R represents H or CH ₃ ).
After culturing in a medium containing (substituted phenyl) valeric acid,
Hydroxybutyric acid and 3-hydroxy-5 represented by the general formula (1) and having R in common with the general formula (2).
A method for biologically producing polyhydroxyalkanoate, comprising extracting a polyhydroxyalkanoate containing (4-substituted phenyl) valeric acid as a monomer unit from the culture.

In the biological production method of polyhydroxyalkanoate described above, it is preferable to use chloroform for extracting the polyhydroxyalkanoate. Similarly, it is also preferable to use hypochlorite for the extraction of the polyhydroxyalkanoate. In addition, the medium is preferably a medium containing either nonanoic acid or lauric acid (dodecanoic acid) in addition to the 5- (4-substituted phenyl) valeric acid represented by the general formula (2). .

[0020]

BEST MODE FOR CARRYING OUT THE INVENTION The OK4 strain, which is a microorganism of the present invention, was obtained as a PVA-assimilating growth microorganism from a soil in Okayama Prefecture using a selective medium containing PVA as a single carbon source.

The OK4 strain is a microorganism having the characteristics summarized in the following Tables 1-3.

[0022]

[Table 1]

[0023]

[Table 2]

[0024]

[Table 3] Based on the above properties, it was confirmed that it was appropriate to identify this strain and OK4 strain as Burkholderia sp.

Further, as described in detail in Examples below, this OK4 strain uses nonanoic acid (NA) and 5- (4-substituted phenyl) valeric acid represented by the above general formula (2) as a substrate ( 3HB, which has not been reported before, and 3-
It has been found that PHA containing hydroxy-5- (4-substituted phenyl) valeric acid as a monomer unit can be simultaneously produced. Based on the above characteristic properties, OK4 strain was concluded to be a new strain belonging to the genus Burkholderia, and it was concluded that Burkholderia sp.
urkholderia sp.) Deposited as OK4 strain with the Institute of Biotechnology and Industrial Technology, Ministry of International Trade and Industry (Deposit No .: FER)
M P-17371).

When the OK4 strain of the present invention is grown normally, a natural medium for general growth (such as LB medium) or M9
A medium obtained by adding about 0.2% to 0.5% of yeast extract, peptone, or the like to an inorganic salt medium such as a medium can be used.

On the other hand, when PHA is accumulated, a fatty acid having about 7 to 12 carbon atoms, for example, nonanoic acid (NA) or octanoic acid (OA) is used as a growth carbon source in an inorganic salt medium such as M9 medium. Decanoic acid, lauric acid (dodecanoic acid), medium chain fatty acids of about 0.1% to 0.2%, and 5- (4-substituted phenyl) valeric acid represented by the general formula (2) as 0.1%. Using a medium to which about 1% to 0.2% is added,
The cells are cultured from the late logarithmic growth phase to the stationary phase, and the cells are harvested by centrifugation or the like. Then, the cells were added to an inorganic salt medium in the absence of a nitrogen source by adding 0.1% to 0.2% of medium-chain fatty acids.
% And 5- (4-substituted phenyl) valeric acid represented by the general formula (2) is further cultured in a medium to which about 0.1% to 0.2% is added.

Alternatively, from the beginning, the nitrogen source concentration in the inorganic salt medium is limited to about 1/10, and the medium-chain fatty acid is reduced to 0.1%.
About 1% to 0.2%, and 5-
0.1% to 0.2% of (4-substituted phenyl) valeric acid
It is also possible to employ a method in which the cells are cultured in a medium supplemented to a certain degree, the cells are harvested at the time from the late logarithmic growth stage to the stationary phase, and the desired PHA is extracted.

For such cultivation, various methods for culturing microorganisms, such as a batch type, a flow batch type, a continuous culture type and a reactor type, can be used.

As described above, the inorganic salt medium used for accumulating PHA is capable of growing microorganisms such as a phosphorus source (usually phosphate) and a nitrogen source (usually ammonium salt or nitrate). Any material may be used as long as it contains the components, and examples thereof include an MSB medium and an M9 medium. Table 4 shows the composition of the M9 medium used in the examples of the present invention.

[0031]

[Table 4] In the biological production method of polyhydroxyalkanoate of the present invention, the target P
The most convenient means for recovering HA is chloroform extraction, which is usually performed. For example, in an environment where it is difficult to use an organic solvent, by treating with a surfactant such as SDS, enzymatic treatment with lysozyme or the like, or treating with a chemical such as EDTA, sodium hypochlorite, or ammonia, intracellular components other than the target PHA can be obtained. , A method of recovering PHA can be adopted. Above all, means using sodium hypochlorite is preferably used.

[0032]

The present invention will be described below in more detail with reference to specific examples. However, the present invention, by these examples,
It is not limited at all.

Example 1 Production of PHA Containing 3HB and 3HPV-1 O grown on M9 agar medium containing 0.1% nonanoic acid
The K4 strain colony was isolated from nonanoic acid 0.1%, PVA (5-
M9 liquid medium containing 0.1% of phenylpentanoic acid) (2
00 mL) and cultured at 30 ° C. After 36 hours, cells were harvested by centrifugation and nonanoic acid 0.1
% And PVA 0.2%, and resuspended in M9 liquid medium (200 mL) containing no nitrogen source (NH ₄ Cl), followed by shaking culture at 30 ° C. for 36 hours. The cells were harvested again by centrifugation, washed once with cold methanol and lyophilized.

After weighing the lyophilized pellet, 10
The suspension was suspended in 0 mL of chloroform and stirred at 60 ° C. for 20 hours to extract PHA. After the extract was filtered through a 0.45 μm filter, the extract was concentrated with an evaporator, and the concentrate was reprecipitated in cold methanol to obtain a polymer. The polymer was dried under reduced pressure at room temperature and weighed.

The composition of the obtained PHA was analyzed as follows. That is, about 10 mg PHA is put in a 25 mL eggplant-shaped flask, dissolved in 2 mL of chloroform,
Add 2 mL of methanol solution containing 3% sulfuric acid and add
The mixture was reacted for 3.5 hours while refluxing at ℃. After the reaction,
After adding 10 mL of deionized water and shaking vigorously for 10 minutes, the lower chloroform layer separated into two layers was taken out and dehydrated with magnesium sulfate. This chloroform layer was subjected to gas chromatography-mass spectrometry (GC-M
S, Shimadzu QP-5050, EI method) to identify the methyl esterified product of the contained PHA monomer unit. Table 5 shows the identified monomer units and their contents.

[0036]

[Table 5] CDW: cell dry weight (dry cell weight) PDW: Polymer dry weight (polymer dry weight) C / P: polymer dry weight / dry cell weight 3HB: 3-hydroxybutyric acid 3HV: 3-hydroxyvaleric acid 3HHp: 3- Hydroxyheptanoic acid 3HN: 3-hydroxynonanoic acid 3HPV: 3-hydroxy-5-phenylvaleric acid (Example 2) Production of PHA containing 3HB and 3HTVA (3-hydroxy-5- (p-tolyl) valeric acid) Macromolecules, 29, p.1762-1766 (1996) and Macr
omolecules, 27, pp. 45-49 (1994), according to the Grignard reaction.
(p-tolyl) valeric acid: 5- (p-tolyl) pentanoic acid)
Was synthesized. That is, 5-bromovaleric acid was dissolved in anhydrous THF (tetrahydrofuran), and a 3M methylmagnesium chloride THF solution was added dropwise at −20 ° C. under an argon atmosphere. After stirring for about 15 minutes, a THF solution of p-bromotoluene and magnesium was further added dropwise, and then a THF solution of 0.1 M Li ₂ CuCl ₄ was added (the temperature was kept at −20 ° C.). The reaction solution was returned to room temperature and further stirred overnight. Thereafter, this solution was poured into an ice-cooled 20% aqueous sulfuric acid solution and stirred, and an aqueous layer was collected. The resulting aqueous layer was saturated with brine and extracted with ether. Further, the ether extract was extracted with 100 mL of deionized water to which 50 g of potassium hydroxide was added, and then the aqueous layer was acidified with a 20% aqueous sulfuric acid solution to collect an oily portion.

This oily part was dried over magnesium sulfate and then dried under reduced pressure. When the obtained product was dissolved in deuterated acetone and measured by ¹ H-NMR, Macromolecules, 29, p. 1762-1766 (199) in the original paper disclosing the synthesis method was obtained.
This was consistent with the TVA data shown in 6). This product was used as a TVA substrate in the following experiments.

Colonies of the OK4 strain grown on an M9 agar medium containing 0.1% nonanoic acid were isolated from 0.1% nonanoic acid,
The cells were inoculated into an M9 liquid medium (200 mL) containing 0.1% of TVA, and cultured at 30 ° C. After 36 hours, cells were harvested by centrifugation, nonanoic acid 0.1%, TVA 0.2
% M9 liquid medium (200 mL) without a nitrogen source (NH ₄ Cl) containing
The cells were cultured with shaking for a time. The cells were harvested again by centrifugation, washed once with cold methanol and lyophilized.

After weighing the lyophilized pellet, 10
The suspension was suspended in 0 mL of chloroform and stirred at 60 ° C. for 20 hours to extract PHA. After the extract was filtered through a 0.45 μm filter, the extract was concentrated with an evaporator, and the concentrate was reprecipitated in cold methanol to obtain a polymer. The polymer was dried under reduced pressure at room temperature and weighed.

The composition of the obtained PHA was analyzed as follows. That is, about 10 mg PHA is put in a 25 mL eggplant-shaped flask, dissolved in 2 mL of chloroform,
Add 2 mL of methanol solution containing 3% sulfuric acid and add
The mixture was reacted for 3.5 hours while refluxing at ℃. After the reaction,
After adding 10 mL of deionized water and shaking vigorously for 10 minutes, the lower chloroform layer separated into two layers was taken out and dehydrated with magnesium sulfate. This chloroform layer was subjected to gas chromatography-mass spectrometry (GC-M
S, Shimadzu QP-5050, EI method) to identify the methyl esterified product of the contained PHA monomer unit. Table 6 shows the identified monomer units and their contents.

[0041]

[Table 6] CDW: cell dry weight (dry cell weight) PDW: Polymer dry weight (polymer dry weight) C / P: polymer dry weight / dry cell weight 3HB: 3-hydroxybutyric acid 3HV: 3-hydroxyvaleric acid 3HHp: 3- Hydroxyheptanoic acid 3HN: 3-hydroxynonanoic acid 3HTVA: 3-hydroxy-5- (p-tolyl) valeric acid (Example 3) Production of PHA containing 3HB and 3HPV-2 M9 agar medium containing 0.1% lauric acid A colony of the OK4 strain grown on the above was lauric acid 0.1%, PVA
The cells were inoculated into an M9 liquid medium (200 mL) containing 0.1% of (5-phenylpentanoic acid) and cultured at 30 ° C. 36
After an hour, the cells were harvested by centrifugation, and nonanoic acid was added to the cells.
Resuspend in M9 liquid medium (200 mL) containing 1%, PVA 0.2% and no nitrogen source (NH ₄ Cl),
Further, shaking culture was performed at 30 ° C. for 36 hours. This cell is
Harvested again by centrifugation, washed once with cold methanol and lyophilized.

After weighing the lyophilized pellet, 10
The suspension was suspended in 0 mL of chloroform and stirred at 60 ° C. for 20 hours to extract PHA. After the extract was filtered through a 0.45 μm filter, the extract was concentrated with an evaporator, and the concentrate was reprecipitated in cold methanol to obtain a polymer. The polymer was dried under reduced pressure at room temperature and weighed.

The composition of the obtained PHA was analyzed as follows. That is, about 10 mg PHA is put in a 25 mL eggplant-shaped flask, dissolved in 2 mL of chloroform,
Add 2 mL of methanol solution containing 3% sulfuric acid and add
The mixture was reacted for 3.5 hours while refluxing at ℃. After the reaction,
After adding 10 mL of deionized water and shaking vigorously for 10 minutes, the lower chloroform layer separated into two layers was taken out and dehydrated with magnesium sulfate. This chloroform layer was subjected to gas chromatography-mass spectrometry (GC-M
S, Shimadzu QP-5050, EI method) to identify the methyl esterified product of the contained PHA monomer unit. Table 7 shows the identified monomer units and their contents.

[0044]

[Table 7] CDW: cell dry weight (dry cell weight) PDW: Polymer dry weight (polymer dry weight) C / P: polymer dry weight / dry cell weight 3HB: 3-hydroxybutyric acid 3HHx: 3-hydroxyhexanoic acid 3HO: 3- Hydroxyoctanoic acid 3HD: 3-hydroxydecanoic acid 3HDD: 3-hydroxydodecanoic acid 3HPV: 3-hydroxy-5-phenylvaleric acid

[0045]

According to the present invention, 3HB and 3-hydroxy-5 are used.
Barkholderia species OK4 (FERM P-17371) is provided as a novel strain having the unique property of simultaneously producing PHA using-(4-substituted phenyl) valeric acid as a monomer unit. Is used as a substrate and cultured under appropriate conditions in a medium containing 5- (4-substituted phenyl) valeric acid to obtain 3HB and 3-HB.
This makes it possible to simultaneously produce PHA using hydroxy-5- (4-substituted phenyl) valeric acid as a monomer unit. In the method of the present invention, in addition to PHA having 3-hydroxy-5- (4-substituted phenyl) valeric acid as a monomer unit, PHA having 3HB as a monomer unit at the same time
However, there is an advantage that it can be easily produced in a high yield.

Claims (5)

[Claims] 1. 3-hydroxybutyric acid and the following general formula (1): (Wherein R represents H or CH ₃ ) A novel microorganism Burkholderia species simultaneously producing polyhydroxyalkanoate containing 3-hydroxy-5- (4-substituted phenyl) valeric acid as a monomer unit (Burkholderia sp.) OK4 strain (FERM P-17371). 2. The Burkholderia according to claim 1, wherein
Burkholderia sp. OK4 strain (FERM P-173)
71) with the following general formula (2): (Wherein R represents H or CH ₃ ).
After culturing in a medium containing (substituted phenyl) valeric acid,
Hydroxybutyric acid and 3-hydroxy-5 represented by the general formula (1) and having R in common with the general formula (2).
A method for biologically producing polyhydroxyalkanoate, comprising extracting a polyhydroxyalkanoate containing (4-substituted phenyl) valeric acid as a monomer unit from the culture. 3. The method according to claim 2, wherein chloroform is used for extracting the polyhydroxyalkanoate.
The biological production method of the polyhydroxyalkanoate according to the above. 4. The method according to claim 2, wherein hypochlorite is used for the extraction of the polyhydroxyalkanoate.
The biological production method of the polyhydroxyalkanoate according to the above. 5. The method for producing polyhydroxyalkanoate according to claim 2, wherein the medium contains either nonanoic acid or lauric acid.