WO1992022659A1 - Procede de production d'un ester d'acide organique polyhydroxy - Google Patents

Procede de production d'un ester d'acide organique polyhydroxy Download PDF

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Publication number
WO1992022659A1
WO1992022659A1 PCT/JP1992/000751 JP9200751W WO9222659A1 WO 1992022659 A1 WO1992022659 A1 WO 1992022659A1 JP 9200751 W JP9200751 W JP 9200751W WO 9222659 A1 WO9222659 A1 WO 9222659A1
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WIPO (PCT)
Prior art keywords
organic acid
pha
acid ester
cells
polymer
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PCT/JP1992/000751
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English (en)
Japanese (ja)
Inventor
Koichi Mochida
Yoshikazu Kondo
Masao Matsui
Kunio Ichihashi
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Kanebo, Ltd.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from JP3324953A external-priority patent/JP2925385B2/ja
Application filed by Kanebo, Ltd. filed Critical Kanebo, Ltd.
Priority to JP4353197A priority Critical patent/JPH05336982A/ja
Publication of WO1992022659A1 publication Critical patent/WO1992022659A1/fr

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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
    • C12N1/06—Lysis of microorganisms
    • 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
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters
    • C12P7/625—Polyesters of hydroxy carboxylic acids

Definitions

  • the present invention relates to a microbiological production method of a polyhydroxy organic acid ester using a microorganism of the genus Drosophila, and a polyhydroxy organic acid ester.
  • the present invention relates to a method for separating and purifying a polyhydroxy organic acid ester from an accumulating bacterial body.
  • A) is biodegradable and has been expected to be used in various fields as a material for molded resins, pharmaceutical preparations, medical equipment, etc.
  • JP-A-57-74084 discloses the species Alkaligenes (A. faecalis, A. rulan).
  • Japanese Patent Application Laid-Open No. 57-15393 discloses an A.U.T. port, while A.A.Lass, A.Aqua Marine, and A.U.
  • Japanese Patent Application Laid-Open No. Sho 59-220192 discloses Nokajia, Azotobakyu, Noshiras, Micrococcas, Resodium, Rhodos Pirinolem, methylonacteria, pseudomonas, and hydrogenomas are disclosed in Japanese Patent Application Laid-Open No. 60-199392, Alkali Genippo Les.
  • JP-A-60-2148 discloses the genus Alcaligenes
  • JP-A-60-25119 discloses azotobacter monovinenlandi.
  • mutant strain No. 85 discloses an all-force regenerative unit mouth face
  • Japanese Unexamined Patent Publication No. Sho. 19-91 discloses a genus Protomonas. Paktor's Vin Vietnamese Landy, Alca Ligenes Utrofas, Zooglare Ramigera, Nautiles Megadica, Special Features JP-A-63-226291 describes pseudomonas oleobolans species, respectively.
  • Poly- ⁇ -hydroxybutyric acid has been found as a cell accumulation of microorganisms belonging to the red sulfur-free bacterium along with polysaccharides and polyphosphoric acid.
  • dioxan which is an organic solvent other than chlorine-based solvent
  • the temperature of the dioxane solution must be raised to a high temperature of 80 ° C or higher, which leads to workability and practicality such as decomposition and deterioration of ⁇ ⁇ ⁇ . It is something that is missing.
  • Japanese Patent Application Laid-Open No. 63-226291 discloses that bacterial cells are converted into sponge plasty, crushed by sonic vibration treatment, and then centrifuged. It describes a method for separating the top layer (PHA) to be separated into 10 layers. In this method, the viscosity of the PHA-containing liquid is still significantly increased by the sonic vibration treatment. In addition, since the floated PHA is separated, impurities having the same specific gravity as PHA cannot be removed.
  • the present invention provides efficient biodegradation by microorganisms that have a high production rate, a large amount of bacteria 15 accumulated in the body, and are safe and easy to handle.
  • To provide a method for the production of a functional polymer to use less organic solvent, to suppress a significant viscosity increase of the PHA-containing liquid, and to add a flocculant.
  • the purpose of the present invention is to provide a method for separating and purifying PHA of high purity which does not require such extra steps.
  • the present invention provides a PHA characterized in that a microorganism belonging to the genus Rho 0 d0 bacter is cultured under aerobic conditions and PHA is accumulated in the cells.
  • This is a method for separating and purifying PHA from PHA-accumulating cells, and adding a lytic enzyme to the cell suspension to lyse the cell wall. After that, the process of separating, collecting and collecting the ⁇ ⁇ ⁇ granules with a particle size of 11 m or more covered with the granule coat present in the cytoplasm is performed.
  • the present invention also relates to a method for separating and purifying PHA, which comprises a step of removing a condylar capsule by treatment with a protease.
  • the present inventors have conducted intensive studies to achieve the above object, and as a result, used microorganisms belonging to the genus Oral genus that are non-pathogenic, and therefore safe and easy to handle.
  • the rate of growth of organisms and the rate of accumulation of PHA in the cells are extremely low, and are extremely high under aerobic conditions (oxygen-containing conditions) that have never been adopted before. It has been found that accumulation can be performed effectively and that the culture can be effectively performed by one-step culture or two-step culture.
  • the pH of the culture broth was found to be less than 4.5 in the microbiological production of PHA using a microorganism belonging to the genus Rhodopacter.
  • the consumption of PHA accumulated in the cells increases, the addition of yeast to the medium increases the accumulation of PHA, and it is mainly used as a carbon source for medium components.
  • PHA production can be effectively performed by using nits.
  • the lytic enzyme to lyse only the cell wall without damaging the PHA granule membrane, PHA is made into granules more easily than other cellular components. It has been found that they can be separated and that the membrane components can be easily removed from the PHA granules.
  • the present invention has been completed based on the above-mentioned many findings. Next, the method for producing the PHA of the present invention will be described.
  • microorganisms belonging to the genus Mouth Dactor used in the present invention belong to the genus Mouth Dacta which can accumulate PHA in the cells by aerobic cultivation. Any microorganism can be used.
  • mouth cavities can be used.
  • the process is performed until the bacterial concentration is at least 10 ml.
  • a two-step method in which the bacteria are grown and the subsequent production of the polymer of the present invention by nitrogen starvation culture is possible.
  • a medium is initially mixed with a nitrogen source and PHA is accumulated simultaneously with the growth. .
  • PHA is accumulated simultaneously with the growth.
  • yeast means a yeast cell, a dead yeast cell, or a yeast ex.
  • the yeast to be added is preferably yeast ex.
  • the amount of yeast added is usually 0.1 to 3 g Z ⁇ , more preferably 0.5 to 2 g / &, and 5 is usually 0.1 to 1 g for yeast yeast. 2 / H, preferably between 0.2 and 1.5 g / &
  • the bacteria are separated from the culture solution.
  • any of conventionally known methods known in the art may be used.
  • freeze-dried cells, hot-air-dried cells, acetate or ⁇ is dehydrated bacterial cells by methanol.
  • a non-polar solvent is used as a non-polar solvent, adding a porcine mouth to elute ⁇ ⁇ ⁇ Thus, a residue consisting of ⁇ ⁇ ⁇ is obtained.
  • PHA may be used without extracting PHA from the cells, but it may be used with each cell. If the PHA granules in the cells are large, it is advantageous to carry out separation / purification by the PHA separation / purification method of the present invention described later in detail.
  • Such a PHA may be, for example, a poly-3-hydroxybutyrate ester (hereinafter referred to as PHB) represented by the following formula UU.
  • PHB poly-3-hydroxybutyrate ester
  • Qualitative analysis of this structure can be performed by IR, NMR, etc.
  • the property to heat is DSC, It is possible by TMA etc.
  • the molecular weight of the polymer can be measured by the viscosity method, GPC method, etc.
  • the biodegradability is determined by molding the polymer into a film or the like and embedding it in the soil.-The physical properties (strength, elongation), shape (surface, cross section) and molecular weight of the polymer during a period It becomes possible by measuring changes and the like.
  • Polymers according to the present invention include organic solvents, such as, for example, quinolone, dimethylamine, 1,2-dichloronorethane, and
  • the PHA-accumulating bacteria used belong to the genus Oral genus, the genus Galligenes (A1ca1igenes) or the genus Bacillus (Baci11us). Any microorganism can be used as long as it is a microorganism that accumulates PHA in the cells.
  • the microbial you accumulate in the body, P H A is exists follicles or are in the intracellular granulosa and granules • S).
  • PHA granule membrane is composed of proteins such as PHA synthase and PHA-degrading enzyme and a small amount of phospholipids.It isolates PHA from the cytoplasm in the cells, and forms granules in a stable form. Work to keep
  • the particle size of the PHA granules in the cells should be at least 0.1 ⁇ m. Separation / recovery is remarkable if the particle size is less than 0.11 m However, even if PHA granules having a fraction of less than 0.1 ⁇ m and a fraction of less than 0.1 were excluded and PHA granules having a fraction of less than 0.1 ⁇ m were excluded, there is no practical problem. Before performing the separation operation, confirm that it is at least 0.1 ⁇ m by microscopy. As described above, the size of PHA granules that accumulate in microorganisms is preferably 0.1 Mm or more, and granules smaller than 0.1 ⁇ m are excluded during the separation and purification process. Possibilities ⁇ there is.
  • the lytic enzyme used in the present invention digests the cell wall, but may be anything as long as it does not digest the granule membrane. As such an enzyme, for example, lysozyme is not exposed, and a lytic enzyme is added to an aqueous solution in which the cells are suspended.
  • a commonly used microfiltration membrane or ultrafiltration membrane can be used.
  • polystyrene, pollip, film, etc. such as propylene, polytetrafluoroethylene, polycarbonate, etc.
  • a specific hole is formed in the plane.
  • the pore size of the filtration membrane is usually at least 0.05 m or more. Combination of centrifugation and filtration can also be achieved efficiently or with high precision.
  • the PHA granules thus separated are treated with a protease to dissolve and remove the PHA granule membrane. That is, the PHA granules are resuspended in pH 6 to 8 (1! To 100 M Tris-HC1 buffer or phosphate buffer), and the protease is reduced to 100 to 100 mM. l Add 0 g Z ml and incubate at 30-4 G ° C for 30-60 minutes.
  • proteolytic enzymes used are not particularly limited, but include, for example, pronase, Nagase, biolase, and papain trypsin.
  • the throat is broken. .
  • PHA is separated by centrifugation or the like. After the reaction, wash with water if necessary.
  • the processing time can be reduced as the processing area increases.
  • the PHA granules thus formed may be left as it is or may be chopped and added to a buffer in which a protease has been dissolved, and reacted in the same manner as described above to obtain a protein. Dissolve and remove.
  • the PHA can be separated by the above-described separation operation after the proteolytic enzyme treatment is performed first as described above. It is. Proper treatment with proteolytic enzymes keeps PHA granules in a granular form even after the membrane has been dissolved, making subsequent separation operations easy. Power.
  • sonication and sonication using about ⁇ ⁇ of audible sound waves may be performed. it can .
  • the treatment is performed to such an extent that the granules themselves are not decomposed.
  • cool the PHA dispersion Depending on the type of the bacterium, the treatment conditions naturally change, but a sonic wave or a supersonic wave having a frequency of IflKHz to KHz is used. These frequencies are optional, depending on the type of microorganism. Strength is usually rather then preferred 0.
  • FIG. 2 is a graph showing the measurement results of the thermal analysis of the polymer obtained in Example 4.
  • FIG. 4 is a chart showing the results of 13 C-NMR spectrum analysis of the polymer obtained in Example 4.
  • BEST MODE FOR CARRYING OUT THE INVENTION The method for producing PHA of the present invention and the method for separating and purifying PHA from cells accumulating PHA of the present invention will be described based on Examples. However, the invention is not limited to only the originally related embodiments.
  • the washed cells were added to one liter of the starvation culture medium, mixed, and suspended. 1G Qm1 was dispensed into 10 50Qmi Yosaka loflascos at a time, and shaking culture was performed at ⁇ ° C for 8 days.
  • the cultured cells were collected by centrifugation under the same conditions as above, added with 1 ml of acetate, suspended, and allowed to stand for 10 minutes, then centrifuged at 300 fl G, 5 The cells were collected under the condition of minutes. This was repeated four times to dehydrate the cells. Acetone adhering to the cells was volatilized by air drying.
  • Chloroform was distilled off under reduced pressure at 35 ° C.
  • the film-like polymer formed in the flask was washed using methanol, acetone, xanthane and ether in this order.
  • the cultivation was carried out in the same manner as in Example 1 except that the nitrogen source and its content were changed.o
  • the nutrient source was ammonium sulfate. Cultivation was carried out with a ratio of nitrogen source to carbon source, which is nourishment of microorganisms, that is, C / N was about 1.8- ⁇ .
  • Table 1 shows the amount of the polymer accumulated in the body per dried cell of the obtained cells. The fact that nitrogen-starved culture is not performed is apparent from the fact that the residual amount is indicated by the measured value of the nitrogen source after the culture, which is apparently t5 '.
  • the number of viable cells after culturing was measured by the plate dilution culture method (using YPS agar medium). As a result, the number of viable cells was almost the same as before cultivation, and was almost the same regardless of the dalkos concentration. It was shown, (2. 7 X 7. 9) X 10 7 / ml were Tsu der. Two days after starvation culture, when observed with a phase-contrast microscope, the live cells showed that polymer granules had accumulated in the cells, and 3 to 5 particles per cell Met. Further cultivation is continued.
  • the morphology of the cells is spherical (about 1 m in diameter) and rugby ball-like (short diameter l «m, long diameter several ⁇ ). Change until close to All the cells had been filled with PHA granules. Table 2 shows the results.
  • the cells were collected by centrifugation under the same conditions as above, and acetonitrile was added. , And the mixture was allowed to stand for 10 minutes, and then centrifuged at 3000 G for 15 minutes to collect the cells. The cells were dehydrated by repeating this three times. Acetone adhering to the cells was volatilized by air drying.
  • the film-like polymer remaining in the flask is washed using methanol, acetone and xanthate and ether in this order. Thereafter, it was dried, dissolved again in a black hole hole, and once filtered to obtain a 5-hole black hole solution. The chloroform was distilled off to give an ester. The same purification operation was repeated three times to obtain PHA containing PHB as a main component.
  • Mouth protein cavitates ATC C11166 was added to yeast extract at 1 g / without or without yeast cultivation, and starvation culture was performed under the same conditions as in Example 1. Yeast ex The product made by Ku was used. After culturing at 32 ° C for 86 hours, PHA was obtained in the same manner as in Example 1.
  • the starvation medium contains sufficient amounts of thiamine, biotin, and nicotinic acid, which are essential for the growth of oral protein cavities, ATCC 11166.
  • yeast yeast is not a commonly known addition for growth.
  • the addition of yeast ex- traction performed here increased PHA accumulation rather than growth.
  • the PHA was cultured by changing the ratio of the yeast extract (manufactured by Nacalai Tesque) added to the nitrogen-starved medium. Yes. The results are shown below.
  • the amount of PHA produced without yeast excretion was O.g per liter of culture medium and 0.5 g of yeast excretion / 1 / It is 10 or less.
  • Example 3 Example 3 was repeated, except that instead of yeast excretion, Ebbose tablets (trade name, Tanabe Seiyaku Co., Ltd.) were used as powdered milk. Then, culture was performed. Table 4 shows the results.
  • ATCC 11166 In the same manner as in Example 1, the cells were transplanted into .300 ml of YPS medium and cultured to obtain -washed cells. A suspension was prepared by adding 7 ml of physiological saline thereto, and 2 ml of each suspension was taken and transferred to a nitrogen starvation medium described below.
  • the nitrogen-starved medium had the composition described in Example 1 and added yeast yeast (manufactured by Difco) to lgZ.
  • the pH was 4.7, 6.0, and 7.0. No. 3 were prepared, and 50 ml of each medium was prepared in ⁇ !
  • each ⁇ ⁇ is adjusted to 4.4, 4.6 to 4.7 and 6.0 to 6.3 using 1N hydrochloric acid, and further cultured for 50 hours Spelled. After the culture, the cells were collected, PHA was extracted by the method described in Example 4, and the content in the cells was measured. The results are summarized in Table 5.
  • the PHA of (1) was significantly reduced at pH 4.4, indicating that it was hardly accumulated at 15 hours.
  • the concentration was% ⁇ 24 ⁇ 6 6 4.6 to 7 4.7.
  • the pH was set to 6.6 to 7.0 for 15 hours in (3) and then to 6.Q to 6.3 for 50 hours thereafter,% capacity was accumulated.
  • the cell yield after culturing was not much different.
  • Table 5 First 15 o'clock Next 50 h Intracellular PHA Dried cell mass pH after pH (%) (g / l)
  • IFO 12203 was respectively transplanted to 200 ml of YSP medium in the same manner as in Example 4. After culturing for 48 hours at 27 ° C, cells were collected and washed.
  • the carbon source for the starvation culture was prepared in mannit gZ, yeast extract (manufactured by Nakarai Tesque) 1 / H, and washed into the medium 200 ml described in Example 4 prepared in Example 4.
  • the cells were transplanted and cultured with shaking at 27 ° C for 90 hours.
  • the cells were collected, and PHA was extracted in the same manner as in Example 4, and the intracellular content was measured. The results are shown in Table 6.
  • Figure 1 shows the molecular weight distribution curve.
  • the magnetic field was 3 G QMHZ, and the solvent was NMR using a deuterated black hole.
  • FIG. 3 shows the results of the i fi-NMR measurement of the PHA obtained in Example 4, and FIG. 4 shows the results of the 13 C-NMR measurement.
  • the PHA obtained in Example 4 contains hydroxybutyric acid (PHB) and hydroxyvaleric acid (PHV) units, and the ratio thereof is about 95: 5. I knew it was.
  • Example 12 hydroxybutyric acid (PHB) and hydroxyvaleric acid (PHV) units, and the ratio thereof is about 95: 5. I knew it was.
  • Example 2 The PHA recovered in Example 2, EX p. No. 4 and the PHA produced and recovered in Example 4 according to the present invention were dissolved in chloroform and placed on a glass plate. Cast and air-dried film molding. This film was buried in our garden (4-1, Kanebo-cho, Hofu City), and its strength, elongation, and morphological changes over time were observed. The results are shown in Table 7 and show that the disintegration is complete until complete loss of strength in about 60 days. Table7
  • PHA-accumulating bacterial cells cultured in the same manner as in Example 4 (a particle size of 0.4 to ( It was observed that 2-3 granules of 6 were contained.) 5 g (corresponding to lg of dry cells and lg of dry cells) was transferred to a pH 7.0, 50 mM phosphate buffer solution. Suspend at 30 ml and add 50 mg of lysozyme (derived from lysozyme chloride (purified) egg white, manufactured by Nakarai Tesque), 50 mg of 1 M CaC120.5 ml, ⁇ ⁇ ⁇ ⁇ 0.5 mg of Pase (manufactured by Sigma, type I.) was added, and the mixture was stirred and reacted at room temperature for 30 minutes.
  • lysozyme derived from lysozyme chloride (purified) egg white, manufactured by Nakarai Tesque
  • This phenol is cut into a size of about 1 X 1 mm, immersed in pH 7.0, 5 GmM phosphate buffer, and added with 2 Qmg of protease and 8 Q The reaction was performed at ° C for 6 hours. After the reaction, the film was washed with water and dried (D. 5 g).
  • Mouth donocta cap slatus (R. capsulatus ATCC 11166) was cultured in the same manner as in Example 4 and then freeze-dried. (8 G%) was suspended in 100 ml of 50 mM Tris-HCl buffer (pH 8.0) and stirred gently for 5 minutes. As a result of microscopic examination, it was found that the diameter of the PHB granules in the microbial cells was Q.4 to (! .8 // m, one or two viable cells existed in one microbial cell.
  • M MgC 12 1 ml, lysozyme (derived from egg white, lysozyme chloride (purified), Nacalai Tesque) 50 mg, DNAse (Sigma, type) I) 1 mg and 2 ml of Q.l MEDTA sodium were added and stirred to dissolve.After stirring for 30 minutes, the container was transferred to crushed ice and treated at 10 KHz for 2 minutes. Transfer to a centrifuge tube, centrifuge at 5000 G for 20 minutes, remove the supernatant, add 10 ml of 50 mM Tris-HCl buffer (pH 8.0), and suspend the crude PHA suspension. The suspension was centrifuged under the same conditions, and the supernatant was discarded.
  • the method for producing a polyhydroxy organic acid according to the present invention has a higher polymer production efficiency and a higher intracellular accumulation rate than the conventionally proposed methods, and has a higher production efficiency. It has a large characteristic that has not been found in the past, such as high stability of the rimer. In addition, since the intracellular accumulation rate can be extremely high, it is very advantageous for extraction and purification of the polymer. In addition, manufacturing processes such as producing the polymer without the use of photoenergy and with or without nitrogen starvation. However, the conditions are very industrially advantageous as compared with the conventional method. Therefore, its economic value is also very large.
  • high-purity PHA can be safely and efficiently separated and purified from PHA-accumulating bacteria.

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Abstract

L'invention se rapporte à un procédé de production d'un polymère biodégradable, à l'aide d'un micro-organisme qui peut produire un polymère en grande quantité, qui présente un rapport élevé d'accumulation intracellulaire, et qui est sûr et aisément manipulable; à un procédé permettant de séparer et de purifier de manière efficace et sûre un ester d'acide organique polyhydroxy accumulé dans un micro-organisme; à un procédé de production d'un ester d'acide organique polyhydroxy, caractérisé par la culture d'un rhodobacter dans un milieu aérobie afin de produire un polymère comprenant un ester d'acide organique polyhydroxy; et à un procédé de séparation et de purification d'un ester d'acide organique polyhydroxy accumulé dans une cellule microbienne, caractérisé par une étape d'addition d'une enzyme lytique à une suspension cellulaire dans le but de dissoudre les parois cellulaires, une étape de séparation et d'extraction de granules d'ester d'acide organique polyhydroxy présents dans un cytoplasme, lesdits granules étant chacun recouvert d'une membrane granuleuse et présentant un diamètre égal ou supérieur à 0,1 νm, et une étape consistant à éliminer la membrane granuleuse en la traitant avec une protéase.
PCT/JP1992/000751 1991-06-11 1992-06-11 Procede de production d'un ester d'acide organique polyhydroxy WO1992022659A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP4353197A JPH05336982A (ja) 1992-06-11 1992-12-11 ポリヒドロキシ有機酸エステルの分離・精製法

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
JP3/167811 1991-06-11
JP16781191 1991-06-11
JP3324953A JP2925385B2 (ja) 1991-06-11 1991-12-10 ポリヒドロキシ有機酸エステルの製造法
JP3/324953 1991-12-10

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998007879A1 (fr) * 1996-08-20 1998-02-26 Buna Sow Leuna Olefinverbund Gmbh Procede d'obtention de polyhydroxyalcanoates et leur utilisation
WO2004065608A1 (fr) * 2003-01-20 2004-08-05 Kaneka Corporation Procede permettant la collecte de polyhydroxyalcanoate tres pur a partir de cellules microbiennes

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63198991A (ja) * 1987-02-13 1988-08-17 Mitsubishi Rayon Co Ltd ポリ−β−ヒドロキシ酪酸の分離精製法

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63198991A (ja) * 1987-02-13 1988-08-17 Mitsubishi Rayon Co Ltd ポリ−β−ヒドロキシ酪酸の分離精製法

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ARCHIVES OF MICROBIOLOGY, Vol. 155, No. 4, (1991), BRANDL, H. et al., "The Accumlation of Polny-3Hydroxyalkanoates in Rhodobacter-Sphaerides", p. 337-340. *
PRIKL. BIOKHIN MIKROBIOL, Vol. 25, No. 6, 1989, p. 785-789, "Photorophic Bacteria as Producers of Poly-Bets-Hychoxybutuyraxe", Refer to Biosis Number 89103570. KONDRATEVA EN, KRASIL'NIKOVA EN (Russ). *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998007879A1 (fr) * 1996-08-20 1998-02-26 Buna Sow Leuna Olefinverbund Gmbh Procede d'obtention de polyhydroxyalcanoates et leur utilisation
WO2004065608A1 (fr) * 2003-01-20 2004-08-05 Kaneka Corporation Procede permettant la collecte de polyhydroxyalcanoate tres pur a partir de cellules microbiennes
JPWO2004065608A1 (ja) * 2003-01-20 2006-05-18 株式会社カネカ 微生物菌体からの高純度ポリヒドロキシアルカノエートの回収方法
US7393668B2 (en) 2003-01-20 2008-07-01 Kaneka Corporation Method of collecting highly pure polyhydroxyalkanoate from microbial cells

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