[go: up one dir, main page]

WO2008026619A1 - Procede de production de copolymeres d'acide 3-hydroxyalcanoique - Google Patents

Procede de production de copolymeres d'acide 3-hydroxyalcanoique Download PDF

Info

Publication number
WO2008026619A1
WO2008026619A1 PCT/JP2007/066725 JP2007066725W WO2008026619A1 WO 2008026619 A1 WO2008026619 A1 WO 2008026619A1 JP 2007066725 W JP2007066725 W JP 2007066725W WO 2008026619 A1 WO2008026619 A1 WO 2008026619A1
Authority
WO
WIPO (PCT)
Prior art keywords
hydroxyalkanoic acid
acid copolymer
treatment
copolymer
aqueous suspension
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/JP2007/066725
Other languages
English (en)
Japanese (ja)
Inventor
Yousuke Asai
Yoshihisa Dohno
Tsunekata Kobata
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Kaneka Corp
Original Assignee
Kaneka Corp
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
Application filed by Kaneka Corp filed Critical Kaneka Corp
Priority to JP2008532082A priority Critical patent/JPWO2008026619A1/ja
Publication of WO2008026619A1 publication Critical patent/WO2008026619A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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 method for treating or purifying a 3-hydroxyalkanoic acid copolymer produced by a microbial cell.
  • Poly-3-hydroxyalkanoic acid (hereinafter referred to as PHA) is a thermoplastic polyester that is produced and stored as an energy storage substance in cells of many microbial species, and has biodegradability.
  • plastic waste is treated by incineration, landfill, etc. These treatment methods have problems such as global warming and landfill relaxation. For this reason, the recycling system is being promoted with increasing social awareness of plastic recycling.
  • plastic disposal methods cannot be dealt with by incineration, landfilling, and recycling alone, and many of them are still left in nature. is there. Therefore, biodegradable plastics such as PHA are attracting attention because they are taken into the natural material circulation after disposal and the decomposition products become harmful!
  • PHAs which are produced and accumulated by microorganisms in the microbial cells, are expected to have little adverse effect on the ecosystem because they are incorporated into the natural carbon cycle process. Also, in the medical field, it is considered possible to use implant materials that do not need to be collected as drug carriers.
  • PHA produced by microorganisms usually forms granules and accumulates in the cells of the microorganisms, in order to use PHA as plastic, PHA is separated and extracted from the cells of microorganisms. A process is required.
  • the known methods for separating and purifying PHA from microbial cells can be broadly classified as follows: a method of obtaining PHA by crushing or solubilizing and removing cell components other than PHA, and an organic solvent in which PHA is soluble And a method of extracting PHA from bacterial cells using
  • Non-Patent Document 1 discloses that a cell suspension other than PHA is treated with sodium hypochlorite. body A method for solubilizing the components to obtain PHA is described. Although this method is simple as a process, it is expensive because it requires the use of large amounts of sodium hypochlorite. It is also considered unsuitable for practical use because it causes a significant decrease in the molecular weight of PHA. However, oxidants such as sodium hypochlorite are readily available, and treatment with these is a method that can be used for industrial use because the treatment conditions are relatively mild and the process is simple. . Treatment of cultured microorganisms containing PHA with these oxidizing agents can easily degrade cytoplasmic components and reduce protein content to improve PHA purity.
  • Patent Document 1 states that a high-purity PHA can be obtained by treating PHA-containing cells with a basic component and adding hypochlorite.
  • Patent Document 2 states that the treatment with sodium hypochlorite leaves a non-negligible amount of hydrochloric acid in the resin, which is not suitable for practical use.
  • Patent Document 3 describes washing with warm water, etc., and the reduction effect is judged by the amount of chlorine released at 150 ° C by a gas detector tube and a sensory test.
  • Patent Document 4 describes the concentration and temperature conditions of the oxidizing agent used in the treatment. A similar example can be seen in Patent Document 5.
  • Patent Document 6 the microbial cell suspension containing PHA is heat-treated at 100 ° C or higher to destroy the cell structure, and then treated with proteolytic enzyme treatment and phospholipidase treatment.
  • a method for obtaining PHA by solubilizing bacterial cell components other than PHA in combination with hydrogen oxide treatment is described.
  • the protein is denatured and insolubilized by heat treatment, which increases the burden on the next proteolytic enzyme treatment process, and further, the treatment process is complicated and the enzyme is relatively expensive. For this reason, it has disadvantages such as high cost.
  • Patent Document 13 shows an example of agglomeration of microbial cells using a metal salt having a valence of 2 or more and a surfactant. This is an example relating to the flocculation property of microbial cells, and there is no description regarding the point of selectively aggregating only the PHA in the crushed aqueous suspension.
  • Patent Document 14 describes a method of aggregating PHA suspended in water with heat S, a problem of causing a decrease in molecular weight by heating, and a description of the purity of the obtained PHA. There is no problem.
  • Patent Document 24 a method in which pH adjustment and heating are combined
  • Patent Document 25 a method in which heating is performed at a temperature higher than the melting point of PHA and fusion is used at that time
  • Patent Document 26 states that the agglomeration is performed below the melting point, but this temperature is a heating condition of 100 ° C. or higher, and there is no description regarding the purity of the obtained PHA.
  • Patent Document 27 Another example of agglomeration of PHA by stirring below the boiling point in a suspension of hydrophilic solvent and water Although it can be seen (see Patent Document 27), there is no description regarding the point that sufficient PHA purity is not obtained and the resin recovery rate.
  • the “aqueous suspension containing a 3-hydroxyalkanoic acid copolymer” in the present invention can be regarded in a broad sense as a system in which fine particles are suspended in a liquid phase, and includes a suspension in which solid particles are dispersed. It can be considered as a colloidal dispersion system called. Fine particles dispersed in the liquid phase approach each other due to Brownian motion, turbulence, etc., collide and coalesce, and this includes forces such as repulsion by the electric double layer, van der Waalska, and repulsion by adsorbed molecules. It affects the complexity.
  • Patent Document 2 As a method for agglomerating fine particles generally used in such a system, there is a method in which an electrolyte substance is added or the ion concentration in the liquid is increased by changing the pH, and the charged amount of the particles is reduced to agglomerate. It is known (see Non-Patent Document 2). As described in Patent Document 14, the method of obtaining a PHA aggregate by adding a metal salt having a valence of 2 or more causes cell components such as proteins and PHA to aggregate simultaneously, and is a product. The problem is that metal salts remain in the polymer.
  • Patent Document 1 JP 2005-348640 A
  • Patent Document 2 JP-A-7-177894
  • Patent Document 3 Japanese Patent Laid-Open No. 2003-47495
  • Patent Document 4 Japanese Patent Laid-Open No. 2002-306190
  • Patent Document 5 US Patent No. 5110980 Specification
  • Patent Document 6 Japanese Patent Publication No. 04-61638
  • Patent Document 7 Japanese Patent Laid-Open No. 07-177894
  • Patent Document 8 JP-A-07-31488
  • Patent Document 9 JP-T 08-502415
  • Patent Document 10 Japanese Patent Laid-Open No. 2001-046094
  • Patent Document 11 Japanese Patent Laid-Open No. 07-31487
  • Patent Document 12 International Publication No. 2003/091444 Pamphlet
  • Patent Document 13 JP 2001-057880 A
  • Patent Document 14 Special Table 2000-502399
  • Patent Document 15 Japanese Patent Application Laid-Open No. 55-118394
  • Patent Document 16 Japanese Unexamined Patent Publication No. 57-65193
  • Patent Document 17 JP-A 61-035790
  • Patent Document 18 JP-A-4 264125
  • Patent Document 19 Japanese Unexamined Patent Publication No. 63-198991
  • Patent Document 20 Japanese Patent Laid-Open No. 02-69187
  • Patent Document 21 Japanese Patent Laid-Open No. 07-79788
  • Patent Document 22 Japanese Patent Publication No. 10-504460
  • Patent Document 23 Japanese Patent Publication No. 11 511025
  • Patent Document 24 Japanese Patent Laid-Open No. 11 266891
  • Patent Document 25 Japanese National Patent Publication No. 10-504459
  • Patent Document 26 Japanese Patent Publication No. 7-509131
  • Patent Document 27 Pamphlet of International Publication No. 2004/033700
  • Non-Patent Document 1 J. Gen. Microbiology, 1958, Vol. 19, p. 198—209
  • Non-Patent Document 2 New System Chemical Engineering, Fine Particle Engineering, Ohmsha
  • the decomposition treatment of the bacterial cell constituents using an oxidizing agent such as sodium hypochlorite is a relatively simple treatment with an easily available reagent, and suppresses the decrease in molecular weight associated with this treatment.
  • an oxidizing agent such as sodium hypochlorite
  • the first object of the present invention is to provide an aqueous suspension containing a microorganism containing a 3-hydroxyalkanoic acid copolymer, or a cell structure of a 3-hydroxyalkanoic acid copolymer and a microorganism.
  • an oxidizing agent such as sodium hypochlorite
  • the protein content is reduced, the decrease in the molecular weight of the 3-hydroxyalkanoic acid copolymer is suppressed, and the chlorine content is reduced.
  • the second object of the present invention is to improve the purity of an aqueous suspension containing a 3-hydroxyalkanoic acid copolymer produced by a microorganism with good productivity and without causing a decrease in molecular weight.
  • the present invention provides a purification method characterized by recovering a high 3-hydroxyalkanoic acid copolymer.
  • the present inventors have studied a method for treating an aqueous suspension containing a 3-hydroxyalkanoic acid copolymer with hypochlorite, and efficiently controlling the constituents of the cells while suppressing a decrease in molecular weight. We found conditions that could be removed. In addition, the present inventors have found a method for suppressing the residual chlorine amount depending on the processing conditions. Focusing on the fact that the chlorination of PHA cells is due to hypochlorous acid, we have found a method that can reduce the amount of residual chlorine by treating it in a high pH range that suppresses the generation of hypochlorous acid.
  • the components that make up PHA are composed of CH bonds, etc., but paying attention to the form of chlorine attacking this bond is chlorine radicals and chlorinated radicals, and by suppressing these radicals We have also found that the amount of chlorine remaining can be reduced. We also found a means to suppress the decrease in molecular weight by combining the effective chlorine concentration in the system with the temperature conditions. Combining these methods, we have found a method that can effectively reduce the residual amount of chlorine and efficiently decompose and remove bacterial cell components while suppressing molecular weight reduction.
  • the first aspect of the present invention is an aqueous suspension containing a microorganism containing a 3-hydroxyalkanoic acid copolymer, or an aqueous suspension containing a 3-hydroxyalkanoic acid copolymer and a microbial cell component.
  • a 3-hydroxyalkane comprising a step of treating a suspension with an oxidizing agent containing hypochlorite to decompose microbial cell components and a step of neutralizing the remaining oxidizing agent with sulfite.
  • the present invention relates to a method for treating an acid copolymer.
  • the present inventors are composed of an aqueous suspension containing a 3-hydroxyalkanoic acid copolymer, an aqueous solution containing a large amount of a water-soluble protein, and a highly hydrophobic 3-hydroxyalkanoic acid copolymer. Focusing on this point, we conducted an extensive study. Purified 3-hydroxy Alkanoic acid copolymers are inherently hydrophobic polyesters, and contain water-soluble proteins as hydrophilic solids because they contain proteins and saccharides derived from microorganisms that produce S, 3-hydroxyalkane acid copolymers.
  • a small amount of hydrophobic solvent is added to the 3-hydroxyalkanoic acid copolymer.
  • the present inventors have found that the hydroxyalkanoic acid copolymer can be easily separated as a solid from an aqueous solution containing a large amount of water-soluble protein.
  • the second invention is a method for purifying a 3-hydroxyalkanoic acid copolymer produced by a microorganism, wherein a solvent is added to an aqueous suspension containing the 3-hydroxyalkanoic acid copolymer.
  • the present invention also relates to a purification method capable of recovering an aggregate of 3-hydroxyalkanoic acid copolymer as a solid.
  • an aqueous suspension containing a microorganism containing a 3-hydroxyalkanoic acid copolymer, or a bacterial cell constituent of a 3-hydroxyalkanoic acid copolymer and a microorganism From the aqueous suspension containing the above, it is possible to easily decompose the cell constituents without requiring a complicated treatment such as solvent extraction, and to obtain a highly pure copolymer with a low protein content.
  • by setting the temperature and hypochlorous acid concentration conditions to a certain level or less it is possible to suppress the decrease in the molecular weight of PHA, and also to suppress the chlorination reaction to the components of PHA. PHA with low residual chlorine can be obtained.
  • the 3-hydroxyalkanoic acid copolymer is recovered as a solid from an aqueous solution containing a large amount of water-soluble tannic acid. Disappears. The use of a large amount of solvent necessary for extraction and the heating operation for extraction are unnecessary, so it is possible to suppress a decrease in molecular weight, and a highly pure copolymer with low protein content can be obtained. Obtainable. In other words, the 3-hydroxyalkanoic acid copolymer can be purified while maintaining productivity and quality as a polymer material.
  • the method for treating a 3-hydroxyalkanoic acid copolymer according to the first aspect of the present invention is a method for treating a 3-hydroxyalkanoic acid copolymer produced by a microorganism, comprising a 3-hydroxyalkanoic acid copolymer. Treating an aqueous suspension containing microorganisms to be treated or an aqueous suspension containing 3-hydroxyalkane acid copolymer and microbial cell constituents with an oxidizing agent containing hypochlorite; A step of neutralizing the remaining oxidizing agent.
  • the microorganism in the present invention is not particularly limited as long as it is a microorganism that accumulates 3-hydroxyalkanoic acid copolymer (PHA) in cells!
  • PHA 3-hydroxyalkanoic acid copolymer
  • Examples include Alcalige nes, Ralstonia, Pseudomonas, Bacillus, Azotobacter, Nocardia, and ⁇ ⁇ ⁇ Aeromonas. Can be mentioned.
  • Alkali Genes 'lipolytic power ⁇ ⁇ lipolytic a
  • Alkaline Genes' A. latus Aeromonas 'Cabiae (A. caviae), Aeromonas. Hydrophila, Ralstonia' R. eutr opha, etc.
  • the strain is preferred.
  • microorganism used in the present invention a microorganism belonging to the genus Aeromonas is preferable.
  • Aeromonas cerevisiae or Aeromonas noidrophila is preferable.
  • a microbial cell obtained by culturing these microorganisms under appropriate conditions and accumulating a 3-hydroxyalkanoic acid copolymer in the cell is used.
  • the culture method is not particularly limited, and for example, the method described in JP-A-05-93049 can be used.
  • the 3-hydroxyalkanoic acid copolymer in the present invention is a general term for copolymers composed of 3-hydroxyalkanoic acid.
  • the 3-hydroxyalkanoic acid copolymer is not particularly limited, but specific examples include 3-hydroxybutyrate (3 ⁇ ) and other 3-hydroxyl copolymers. Examples thereof include a copolymer with sialic acid, and a copolymer of 3-hydroxyalkanoic acid containing 3-hydroxyhexanoate (3HH). Further, selected from the group consisting of 3-hydroxypropionate, 3-hydroxybutyrate, 3-hydroxyvalerate, 3-hydroxyhexanoate, 3-hydroxyheptanoate and 3-hydroxyoctanoate 2 Copolymers composed of more than one type of monomer are also included.
  • a copolymer containing 3HH as a monomer component for example, a two-component copolymer of 3HB and 3HH (PHBH) (Macromolecules, 28, 4822— 4828 (1995)), or 3HB and 3-hydroxynorelate ( 3HV) and 3HH ternary copolymer (PHBVH) (Patent No. 277757, Japanese Patent Laid-Open No. 08-289797) More preferable from the viewpoint of physical properties of the obtained polyester.
  • the composition ratio of each monomer unit constituting the binary copolymer PHBH of 3HB and 3HH preferably the 3HH unit having composition ratios such as 1 to 99 mol 0/0 is there.
  • composition ratio of each monomer unit constituting the three-component copolymer PHBVH of 3HB, 3HV, and 3HH is not particularly limited.
  • the content of 3HB unit is 1 to 95 mol%
  • 3HV unit It is preferable that the content of !!-96 mol% and the content of 3HH unit be 1-30 mol%.
  • aqueous suspension containing a microorganism containing a 3-hydroxyalkanoic acid copolymer in the first aspect of the present invention means that a microorganism containing a 3-hydroxyalkanoic acid copolymer is suspended in water.
  • Aqueous suspension containing 3-hydroxyalkanoic acid copolymer and microbial cell constituents means that 3-hydroxyalkanoic acid copolymer and microbial cell components It is suspended in water.
  • an aqueous suspension containing a microorganism containing a 3-hydroxyalkanoic acid copolymer can be used as it is for the oxidant treatment.
  • An aqueous suspension containing a 3-hydroxyalkanoic acid copolymer and a microbial cell component is prepared by disrupting a microorganism containing the alkanoic acid copolymer by a physical or chemical method. Therefore, it is desirable that the subsequent decomposition treatment with an oxidizing agent can be carried out efficiently.
  • the physical crushing method is not particularly limited, but a conventionally known French press, homogenizer, X-press, ball mill, colloid mill, DYNO mill, ultrasonic homogenizer.
  • Examples include fluid shearing force such as Zaichi, solid shearing force, and grinding.
  • a chemical disruption method a method using a drug such as an acid, an alkali, a surfactant, an organic solvent, a cell wall synthesis inhibitor, a method using an enzyme such as lysozyme, pectinase, cellulase, and thymolyase.
  • Other methods include a method using a supercritical fluid, an osmotic crushing method, a freezing method, and a dry pulverizing method.
  • a self-digestion method that uses the action of a protease that is possessed by the cell itself, such as esterase, is also a type of disruption method.
  • the suspension contains a protein secreted by microorganisms, a residue of a culture substrate, and a bacterial cell component when disrupted.
  • the water containing these proteins and the like may be dehydrated before the treatment for adding the oxidizing agent described below.
  • the water containing these proteins or the like is not dehydrated in advance.
  • the 3-hydroxyalkanoic acid copolymer can be efficiently recovered.
  • the method of dehydration is not particularly limited, and examples thereof include filtration, centrifugation, and sedimentation. In order to promote these, inorganic salts and polymer flocculants may be added! /.
  • the concentration of the 3-hydroxyalkanoic acid copolymer in the suspension is not particularly limited, but is preferably 500 g / L or less, more preferably 300 g / L or less.
  • water may be dehydrated or water may be newly added.
  • the oxidizing agent used in the present invention is composed of hypochlorite.
  • hypochlorite include sodium hypochlorite, potassium hypochlorite, calcium hypochlorite and the like, with sodium hypochlorite being preferred.
  • Sodium hypochlorite sodium hypochlorite
  • sodium hypochlorite sodium hypochlorite
  • Sodium hypochlorite is widely used in various fields such as pulp bleaching, sterilization and disinfection of pools and water and sewage systems, and industrial wastewater treatment. It is easier to handle than chlorine used for the same purpose.
  • Sodium hypochlorite is generally produced by a chemical reaction that causes sodium hydroxide to absorb chlorine gas, and gradually decomposes in an aqueous solution to release oxygen. This oxygen is said to show a strong oxidizing power. This oxidizing power can decompose the culture substrate residue, bacterial cell constituents, etc. contained in the suspension.
  • the target organic substances may be chlorinated by by-product chlorine.
  • Sodium hypochlorite is known to be degraded at high temperatures and low pH (Japan Soda Industry Association, Soda Handbook, 1998, etc.).
  • concentration distribution of dissolved active chlorine depends on the pH, and it can be seen that the ratio of hypochlorous acid is high in the weak alkaline to acidic range where the pH is 10 or less to about 3!
  • C1 radical The most reactive form of CH bonds constituting organic substances such as hydrocarbons is C1 radical, but this C1 radical is generated by (1) decomposition of sodium hypochlorite with acid. Starting from the C1 molecule, (2) Also generated from dichlorinated oxygen generated from hypochlorous acid (F. Minisci et al., Chem. Ind., 70, 50 (1988)) Etc. are known. In any case, since hypochlorous acid is used as a starting material, it is necessary to control the decomposition of sodium hypochlorite by making the pH of the system alkaline.
  • the pH during the oxidizing agent treatment is preferably 8 or more, more preferably 10 or more, and still more preferably 12 or more. Preferably, the pH is 14 or less.
  • sodium hydroxide, calcium hydroxide, barium hydroxide, ammonia or the like can be used.
  • acid or alkali may be added to the oxidant to adjust the pH.
  • the oxidant treatment is preferably carried out under light-shielding conditions, and is preferably carried out in a non-metallic material apparatus.
  • non-metallic materials include tanks coated with resin materials such as Teflon (registered trademark), tanks made of non-metallic materials such as FPR, and tanks coated with glass-based materials. Can be mentioned.
  • the PHA contained in the suspension has a high meltability, the heat of dissolution and reaction heat generated at the stage of adding an oxidant and the stage of neutralization treatment of the oxidant to be carried out subsequently. The PHA melts due to the heat of neutralization and, in some cases, agglomerates.
  • the cleaning efficiency decreases when PHA cleaning is performed in a later step. Therefore, it is desirable to treat at a low temperature to remove these heats. This is consistent with the method for suppressing the generation of the C1 radical.
  • the temperature during oxidant treatment should be 40 ° C or less, more desirably 30 ° C or less, even more desirably 20 ° C or less, and 10 ° C or less to ensure the effect. Is desirable. Preferably, it is 5 ° C or higher.
  • the oxidative power of hypochlorite contained in the oxidant causes a decrease in the molecular weight of PHA at the same time as the decomposition of the bacterial constituents. It was also found that PHA more suitable for practical use can be obtained by limiting the effective chlorine concentration after the addition of bismuth to a certain concentration and processing time to a certain time.
  • the concentration of hypochlorite, such as sodium hypochlorite is indicated by the effective chlorine concentration.
  • the effective chlorine concentration in the treatment solution should be 10% by weight or less. More desirably, it is 8% by weight or less, and more desirably 7% by weight or less.
  • the treatment time with the oxidizing agent is preferably short, but 10 hours or less is more desirable, more desirably 5 hours or less, and even more desirably 3 hours or less.
  • the effective chlorine concentration represents the amount of chlorine with respect to free iodine represented by the following formula (1).
  • One molecule of effective chlorine is generated from one molecule of sodium hypochlorite. (Fifth edition, description of official food additives).
  • a compound in which the neutralized salt obtained by neutralization becomes a sulfate can be used, for example, sulfites such as sodium sulfite and calcium sulfite. Is mentioned. From the viewpoint of suppressing chlorination, sulfite is particularly preferable.
  • the addition amount of the neutralizing agent is not particularly limited, but is added to the system after the treatment step with the oxidizing agent.
  • an addition amount that is equal to or more than the equivalent amount of the remaining hypochlorite is preferable.
  • 0.3 equivalent or more is preferable with respect to the used hypochlorite, 0.5 equivalent or more is more preferable, and 1 equivalent or more is more preferable.
  • the upper limit is not particularly limited, but may be, for example, about 2 equivalents or less. .
  • the temperature during the treatment is preferably the same temperature as during the oxidant treatment.
  • Chlorine remaining in the PHA after treatment may be chlorine bonded to PHA, chlorine bonded to a cell component such as protein, chlorine remaining as inorganic chloride, and the like.
  • the chlorination of PHA can be reduced by the method described above, but if chlorinated products such as proteins that are more easily chlorinated remain, they will be counted as C1 in PHA, and chlorine ( Hydrochloric acid) will be generated. Therefore, it is necessary to clean the remaining chlorine as chlorinated proteins and inorganic salts.
  • the present inventors have found that the remaining amount of C1 can be reduced by washing PHA after neutralization with a solvent such as alcohol.
  • a solvent such as alcohol.
  • the protein content typified by N content is reduced, which is thought to contribute to washing and removing chlorinated proteins.
  • the solvent acetone, ketone, alcohol, amine, amides, water, etc. can be used. These may be used alone or in combination.
  • you may heat at the time of a process.
  • an appropriate surfactant or enzyme may be added. The residual amount of chlorine can be further reduced by the cleaning effect of these solvents.
  • the PHA after the treatment can be recovered from the aqueous suspension by a method such as centrifugation or the method of the second present invention.
  • the effective chlorine concentration in the system after the addition of hypochlorite is kept below a certain concentration to suppress the decrease in the molecular weight of PHA, and the temperature and pH during the treatment are adjusted.
  • the method for purifying a 3-hydroxyalkanoic acid copolymer of the second invention is a method for purifying a 3-hydroxyalkanoic acid copolymer produced by a microorganism, comprising a 3-hydroxyalkanoic acid copolymer. Add solvent to aqueous suspension to add 3-hydroxyalkanoic acid copolymer Collecting aggregates of coalescence.
  • microorganism and 3-hydroxyalkanoic acid copolymer in the present invention are the same as described above.
  • the second purification method of the present invention is a method for recovering a 3-hydroxyalkanoic acid copolymer produced by a microorganism.
  • “Aqueous suspension containing a 3-hydroxyalkanoic acid copolymer in the present invention” “Is a suspension of a 3-hydroxyalkanoic acid copolymer in which cell components derived from microorganisms and the like have been removed from the surface of the copolymer.
  • the suspension may contain a protein secreted by the microorganism, a residual culture substrate, a microbial component derived from the microorganism, and the like.
  • the aqueous suspension is subjected to a physical disruption treatment, a chemical disruption treatment, or a combination treatment of the microorganisms containing a 3-hydroxyalkanoic acid copolymer. It is preferable that it is obtained by.
  • the physical crushing treatment is not particularly limited.
  • fluid shear force or solid shear such as conventionally known French press homogenizer, X-press, ball mill, colloid mill, DYNO mill, ultrasonic homogenizer, etc. Examples include force and grinding methods.
  • the chemical crushing treatment is not particularly limited.
  • a method using a drug such as an acid, an alkali, a surfactant, an organic solvent, a cell wall synthesis inhibitor, an enzyme such as lysozyme, pectinase, cellulase, and thymolyase.
  • Other methods include a method using a supercritical fluid, an osmotic crushing method, a freezing method, a dry pulverizing method, and the like.
  • an aqueous suspension containing the above-mentioned microorganism containing a 3-hydroxyalkanoic acid copolymer, or a 3-hydroxyalkanoic acid copolymer and the above-mentioned microorganism is preferable to subject this to a process of adding a solvent as described below. The detailed conditions at this time are the same as in the first aspect of the present invention.
  • the 3-hydroxyalkanoic acid copolymer Prior to the treatment for adding the solvent described below, water containing these proteins and the like is dehydrated.
  • the 3-hydroxyalkanoic acid copolymer can be efficiently recovered without dehydrating water containing these proteins and the like in advance. it can.
  • the method of dehydration is not particularly limited, and examples thereof include filtration, centrifugation, and sedimentation.
  • an inorganic salt or a polymer flocculant may be added.
  • the concentration of the 3-hydroxyalkanoic acid copolymer in the suspension is not particularly limited, but is preferably 500 g / L or less, more preferably 300 g / L or less. Water may be dehydrated in order to adjust the concentration of the 3-hydroxyalkanoic acid copolymer, or water may be newly added.
  • the solvent used in the second present invention is not particularly limited.
  • hydrocarbon solvents such as benzene, toluene, hexane, cyclohexane, and methylcyclohexane
  • jetyl ether, tetrahydrofuran, Ether solvents such as diphenyl ether, aniso-nole and dimethoxybenzene
  • Halogenated hydrocarbon solvents such as methylene chloride, black mouth form, black mouth benzene; acetone, methyl ethyl ketone, methyl isobutyl ketone, etc.
  • Ketone solvents include alcohol solvents such as methanol, ethanol, propanol, isopropanol, n-butyl alcohol, tert-butyl alcohol; nitrile solvents such as acetonitrile, propionitol, benzonitrile; ethyl acetate, butyl acetate, etc.
  • Ester solvent ethylene carbonate, Examples thereof include carbonate solvents such as propylene carbonate; amide solvents such as N, N-dimethylformamide and N, N-dimethylacetamide.
  • hydrocarbon solvents such as toluene, hexane, cyclohexane, and ethyl acetate.
  • Ethyl acetate and butyl acetate are more preferred, with butyl acetate being more preferred.
  • the 3-hydroxyalkanoic acid copolymer is aggregated, and the aggregated 3-hydroxyalkanoic acid copolymer is contained.
  • the solvent layer separates from the aqueous layer.
  • the amount of the solvent to be added is not particularly limited, but it is as small as possible in order to take advantage of the second aspect of the present invention that the 3-hydroxyalkanoic acid copolymer is not extracted and dissolved. It is preferable that In addition, the solubility varies depending on the temperature, and further the cohesiveness is affected. When the temperature is high, the PHA melts. Since the meltability is affected by the copolymer composition ratio, the optimum temperature differs depending on the composition. When PHA melts, the aggregates tend to become large. When the aggregates are too large, the entire PHA becomes agglomerated.
  • the amount of addition force ⁇ of the solvent is smaller, and the temperature when the solvent is added is preferably low. .
  • the amount of the solvent added varies depending on the solvent type, but is preferably 10 times or less, more preferably 5 times or less, in terms of the weight ratio with respect to the 3-hydroxyarhydric acid copolymer in the aqueous suspension. is there . Further, it is preferably 0.5 times or more.
  • the temperature at which the solvent is added is 50 ° C. or lower, preferably 40 ° C. or lower, more preferably 30 ° C. or lower, more preferably 20 ° C. or lower. Moreover, it is preferably 5 ° C or higher.
  • the 3-hydroxyalkanoic acid copolymer forms an aggregate.
  • the agglomerates have a size of several hundred ⁇ to several mm.
  • Means for adjusting the pH before adding the solvent is not particularly limited.
  • any alkali metal or alkaline earth metal hydroxide including sodium hydroxide, potassium hydroxide, lithium hydroxide, calcium hydroxide, etc. may be used as long as the pH can be limited to a specific range.
  • Alkali metal carbonates such as sodium carbonate and potassium carbonate
  • alkali metal salts of organic acids such as sodium acetate and potassium acetate
  • alkali metal borates such as borax
  • alkali metal phosphates such as tripotassium phosphate and dipotassium hydrogen phosphate; or ammonia water.
  • sodium hydroxide, sodium carbonate, potassium hydroxide, and aqueous ammonia are preferable from the viewpoint of industrial production and price.
  • the pH to be controlled by addition of alkali is not particularly limited, but pH 7 or higher is preferable pH 8 or higher is more preferable, and pHIO or higher is more preferable. It can be considered that by increasing the pH, highly hydrophobic PHA is negatively charged, and this charging compensates for cohesion. However, if sufficient cohesiveness can be exhibited depending on the amount of solvent added, etc., it can be aggregated even in a low pH region. [0063] Under these agglomeration conditions, the agglomerates can be evenly dispersed in the aqueous suspension, or the agglomerates can be floated or settled.
  • aggregates are evenly dispersed in the washing suspension, it is preferable to collect them by a method such as filtration. It is preferable to collect by a method. When aggregates are settled, a method such as sedimentation recovery is desirable. Centrifugation is an example of a recoverable method regardless of whether the aggregate is dispersed, suspended, or settled.
  • the filtration method is not particularly limited, but a method using Nutsche or the like, or a method such as suction filtration or pressure filtration is desirable.
  • filtration devices with a pressing function such as filter presses, tube presses, plate presses, gauge presses, belt presses, screw presses, disc presses, centrifugal dehydrators, and multi-chamber cylindrical filters can be selected. is there.
  • a continuous type such as a multi-chamber cylindrical filter is desirable.
  • a method for removing aggregates in a continuous filter there are a string method, a screvers method, a pre-coated screvers method, and the like.
  • filter media such as woven fabric, non-woven fabric, metallic mesh, ceramic filter, porous filter media and polymer membrane
  • a filter aid may be used.
  • filter aids include diatomaceous earth, perlite, asbestos, cellulose, carbonaceous filter aid, acid clay, bentonite, and activated carbon.
  • the agglomerates of the present invention are particles of several hundred inches to several millimeters, they can be filtered using woven fabrics and nonwoven fabrics that are widely used, and filter aids such as diatomaceous earth are particularly useful. Even if it is not necessary, the filtration efficiency is high and the industrial productivity is high.
  • the aggregate When the aggregate is floating, it can be recovered by stationary separation.
  • a method of adhering or including fine bubbles in suspended substances such as aggregates is used.
  • the so-called pressurized levitation method in which water in which air is dissolved under pressure is released into suspension water under the atmosphere, bubbles are generated using the suspension as a core, and the levitation force of the bubbles is used, or the electric power in water.
  • the electrolytic levitation method in which bubbles are generated by the solution action is common.
  • chemicals such as a collecting agent may be added.
  • an aqueous suspension containing aggregates is placed in a high-speed rotating field, whereby a centrifugal force exceeding several thousand to several tens of thousands of times of gravity can be applied.
  • force S a cylindrical centrifugal separator that rotates a cylinder and discharges liquid with a low specific gravity outside the cylinder
  • separation that performs separation on an inclined slope of an umbrella stacked in multiple layers. Examples include a plate type (drabal type) and a decanter type separated by a rotating screw conveyor.
  • aggregates can be obtained with a size of several hundred m to several mm, so that membrane separation is used for separation of bacteria and yeast, separation of useful substances produced in bioreactors, etc.
  • membrane separation is used for separation of bacteria and yeast, separation of useful substances produced in bioreactors, etc.
  • a hydrophobic solvent is replaced with a 3-hydroxyalkanoic acid copolymer.
  • the effect of draining suspended water such as cytoplasmic components can also be obtained by contact. Due to this effect, it is possible to obtain an aggregate of 3-hydroxyalkanoic acid copolymer having a low protein content and high purity.
  • the purified 3-hydroxyalkanoic acid copolymer is obtained by washing the aggregated particles of the 3-hydroxyalkanoic acid copolymer collected by a method such as filtration and stationary separation with water.
  • Power S can be.
  • an organic solvent other than water may be used, or a mixture of water and an organic solvent may be used.
  • the pH of the water may be adjusted.
  • the organic solvent for example, methanol, ethanol, acetone, acetonitrile, tetrahydrofuran, ketones, amines and the like can be used.
  • a surfactant or the like may be added to water.
  • a mixture of these solvents and water may be used. Further, for a short period of time, it is possible to improve the cleaning performance by heating or spraying water or these organic solvents as steam.
  • the 3-hydroxyalkanoic acid copolymer can be isolated by washing the aggregate with water or an organic solvent and then drying it.
  • the second purification method of the present invention does not require a method such as distillation separation, and has the economic advantage that it can recover the solvent and reuse it.
  • the copolymer when the copolymer is recovered from the aqueous suspension containing the 3-hydroxyalkanoic acid copolymer, a small amount of a solvent, particularly a hydrophobic solvent, is added to the copolymer.
  • the copolymer can be easily recovered as an aggregate without caloric extraction.
  • hypochlorous acid in which the molecular weight of the copolymer is reduced by combining the first invention and the second invention, that is, after the neutralization treatment, by adding a solvent to form an aggregate.
  • the copolymer can be recovered with high productivity and without the problem of residual chlorine associated with the salt treatment.
  • the 3-hydroxyalkanoic acid copolymer obtained by the method of the present invention preferably has a weight average molecular weight of 1 million or more. More preferably, it is 1.5 million or more.
  • the weight average molecular weight of the 3-hydroxyalkanoic acid copolymer is determined using a GPC system manufactured by SHIMADZU equipped with Shodex K805 L (300 X 8 mm, two connected, Showa Denko). This is a value obtained by analyzing black mouth form as a mobile phase.
  • the 3-hydroxyalkanoic acid copolymer obtained by the method of the present invention has a chlorine content of 200 ppm or less as measured by the chlorometry method (Mitsubishi Kasei Co., Ltd., TOX-10).
  • the force S is preferable, and it is more preferably 10OOppm or less.
  • the 3-hydroxyalkanoic acid copolymer obtained by the method of the present invention has a nitrogen content of 200 ppm or less as measured with a trace nitrogen analyzer TN-110 manufactured by Diasurummenku. It is more preferable that it is lOOppm or less.
  • room temperature means 25 ° C.
  • Purified PHBH (15 mg) obtained by isolation from cells was dissolved in 700 m Thereafter, insoluble matters were removed by filtration (Ultra Free MC, manufactured by Millipore). Using this solution, the molecular weight of the polymer was measured using a SHIMADZU GPC system equipped with Shodex K805L (300 ⁇ 8 mm, two linked, Showa Denko KK) using Kuroguchi Form as the mobile phase.
  • Each sample was measured using a TN-110 trace nitrogen analyzer manufactured by Diainstrument, or CHN-CORDER, MT-5 (manufactured by Yanaco).
  • the analysis was performed by the chlormetry method (Mitsubishi Kasei Co., Ltd., TOX-10—).
  • the obtained culture solution was heated and stirred at an internal temperature of 60 to 80 ° C. for 20 minutes, and sterilized.
  • the resulting sterilized culture solution was treated by high-pressure homogenizer treatment (PA2K type, Niro Soa vi S. P. A).
  • PA2K type Niro Soa vi S. P. A
  • the pH of the system dropped from 12.5 to 9 to 10 or so.
  • After neutralizing the treatment solution (adding 0.6 times equivalent of sodium sulfite to sodium hypochlorite, adding sodium sulfite with 15 wt% aqueous solution, stirring for 30 minutes at room temperature), centrifuge separation (1 at 8000G) 0 minute treatment).
  • This treatment solution was treated with sodium hypochlorite (effective chlorine concentration in the system 5%, room temperature, lhr stirring). With this sodium hypochlorite treatment, the pH of the system dropped from 12.5 to 9 to 10 or so.
  • the nitrogen content was determined to be 330 ppm using a trace nitrogen analyzer. The molecular weight was 1.87 million. It was found that the protein content can be reduced to 330 ppm in terms of nitrogen by combining the method described in Example 1 with high-pressure homogenization treatment.
  • This treatment solution was treated with sodium hypochlorite (effective chlorine concentration in the system 6.5 %, Room temperature, lhr stirring). This sodium hypochlorite treatment lowers the pH of the system from 12.5 to 9-10. Further neutralization was performed (Na sulfite was added in an amount equivalent to 0.6 times that of Na hypochlorite, Na sulfite was added as a 15 wt% aqueous solution, and the mixture was stirred at room temperature for 30 minutes).
  • the dry purified PHBH had a nitrogen content of 300 ppm (carbon / hydrogen / nitrogen simultaneous determination device) and a chlorine content of lOlOOppm (chlormetric method).
  • the molecular weight was 1.14 million. There are many residues of chlorine that can reduce protein to 300 ppm in nitrogen content while maintaining the molecular weight above 100,000.
  • PH is calculated from the PHBH content in the culture solution after neutralization treatment (addition of 0.5 times equivalent of sodium sulfite to sodium hypochlorite, addition of sodium sulfite in 15 wt% aqueous solution and stirring for 60 minutes at room temperature) Ethyl acetate equivalent to the theoretical amount of BH was added and stirred at room temperature for 90 minutes. The obtained aggregate was filtered and dehydrated, and the washing operation of adding pure water was repeated. After washing with water 40 times the theoretical amount of PHBH, the obtained aggregate was dried (dried overnight at 40 ° C. to 45 ° C.).
  • the dried purified PHBH had a nitrogen content of 400 ppm (carbon / hydrogen / nitrogen simultaneous determination device) and a chlorine content of 2800 ppm (chlormetric method).
  • the molecular weight was 2.16 million. While keeping the molecular weight above 1 million, protein can be reduced to 400 ppm in nitrogen content and the residual chlorine can be reduced to 2800 ppm!
  • PHBH theory calculated from the PHBH content in the culture solution after neutralization (adding 0.5 fold equivalent of sodium sulfite to sodium hypochlorite, adding sodium sulfite in 15 wt% aqueous solution and stirring for 60 minutes at room temperature) To the amount, 0.44 times the amount of toluene was added, and the mixture was stirred at room temperature for 60 minutes. The obtained agglomerates were filtered and dehydrated, and the washing operation in which pure water was added was repeated. After washing with 40 times the theoretical amount of PHBH and further with 60 times the amount of methanol for the theoretical amount of PHBH, the resulting aggregate is dried (dried at 40 ° C to 45 ° C overnight). )did.
  • the dried purified PHBH had a nitrogen content of 65 ppm (trace nitrogen analysis) and a chlorine content of lOOppm (chlormetric method).
  • the molecular weight was 1.47 million. While maintaining the molecular weight above 1 million, protein can be reduced to 65 ppm in nitrogen content, and the remaining amount of chlorine can be reduced to lOOppm.
  • the generation temperature of chlorine liberated for this purified PHBH was monitored by the TG-MS method (TG-DTA / MS, manufactured by Agilent Technologies). TG curve measured in the region from room temperature to 500 ° C No change in weight in the temperature range lower than the decomposition temperature of S and PHBH, and chlorine (hydrochloric acid) cannot be detected by MS. Therefore, it was proved that the amount of chlorine (hydrochloric acid) released with the melting of PHBH was extremely small.
  • pure water was added, and after sufficiently stirring with the precipitate layer, further centrifugation (treatment at 8000 G for 10 minutes) was performed. This washing operation was repeated 5 times.
  • PHBH theory calculated from the PHBH content in the culture solution after neutralization treatment (addition of 0.5 times equivalent of sodium sulfite to sodium hypochlorite, addition of sodium sulfite in 15 wt% aqueous solution and stirring for 60 minutes at room temperature) To the amount, 0.44 times the amount of toluene was added and stirred at room temperature for 60 minutes. The obtained agglomerates were filtered and dehydrated, and the washing operation of adding pure water was repeated. After washing with 40 times as much water as the theoretical amount of PHBH, the obtained aggregate was dried (dried overnight at 40 ° C to 45 ° C).
  • the dried purified PHBH had a nitrogen content of 320 ppm (carbon / hydrogen / nitrogen simultaneous determination device) and a molecular weight of 1.47 million.
  • the chlorine content was 1400ppm (chlormetric method). While maintaining the molecular weight above 1 million, the protein can be reduced to 400 ppm or less in terms of nitrogen content.

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

L'invention concerne la décomposition et l'élimination efficaces de composants constituant des cellules microbiennes, ainsi que l'inhibition simultanée de la diminution ou de la réduction de la quantité de chlorure résiduel par les poids moléculaires de copolymères d'acide 3-hydroxyalcanoïque. L'invention concerne un procédé de traitement de copolymères d'acide 3-hydroxyalcanoïque produits par des microbes, consistant à traiter une suspension aqueuse de microbes contenant des copolymères d'acide 3-hydroxyalcanoïque ou une solution aqueuse contenant à la fois des copolymères d'acide 3-hydroxyalkanoïque et des composants constituant des cellules microbiennes au moyen d'un oxydant contenant un hypochlorite, puis à neutraliser l'oxydant résiduel au moyen d'un sulfite. L'invention concerne également un procédé de purification de copolymères d'acide 3-hydroxyalcanoïque produits par des microbes, consistant à ajouter un solvant à une solution aqueuse contenant des copolymères d'acide 3-hydroxyalcanoïque et à récupérer des agrégats de ces copolymères.
PCT/JP2007/066725 2006-08-30 2007-08-29 Procede de production de copolymeres d'acide 3-hydroxyalcanoique Ceased WO2008026619A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2008532082A JPWO2008026619A1 (ja) 2006-08-30 2007-08-29 ３−ヒドロキシアルカン酸共重合体の製造方法

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
JP2006234260 2006-08-30
JP2006-234261 2006-08-30
JP2006-234262 2006-08-30
JP2006234262 2006-08-30
JP2006234261 2006-08-30
JP2006-234260 2006-08-30

Publications (1)

Publication Number Publication Date
WO2008026619A1 true WO2008026619A1 (fr) 2008-03-06

Family

ID=39135901

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2007/066725 Ceased WO2008026619A1 (fr) 2006-08-30 2007-08-29 Procede de production de copolymeres d'acide 3-hydroxyalcanoique

Country Status (2)

Country Link
JP (1) JPWO2008026619A1 (fr)
WO (1) WO2008026619A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025134498A1 (fr) * 2023-12-18 2025-06-26 三菱瓦斯化学株式会社 Procédé de production d'agrégat d'acide polyhydroxyalcanoïque

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5110980A (en) * 1989-04-06 1992-05-05 Ecole Polytechnique Separation of poly-β-hydroxyalkanoic acid from microbial biomass
JPH11511025A (ja) * 1995-08-21 1999-09-28 ザ、プロクター、エンド、ギャンブル、カンパニー バイオマスからのポリヒドロキシアルカノエートの溶媒抽出
JP2002262886A (ja) * 2001-03-14 2002-09-17 Inst Of Physical & Chemical Res 短鎖脂肪酸由来のモノマーからなる共重合ポリエステルを合成する植物及びポリエステルの製造方法
JP2002306190A (ja) * 2001-04-10 2002-10-22 Canon Inc 生体細胞からのポリ−３−ヒドロキシアルカン酸の分離・回収方法
JP2002543794A (ja) * 1999-05-12 2002-12-24 メタボリックス，インコーポレイテッド ポリヒドロキシアルカノエートを精製するための方法
WO2005085461A1 (fr) * 2004-03-04 2005-09-15 Kaneka Corporation Methode de degradation de l'acide nucleique et son utilisation
JP2005348640A (ja) * 2004-06-09 2005-12-22 Asahi Breweries Ltd Ｐｈａ精製方法

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5110980A (en) * 1989-04-06 1992-05-05 Ecole Polytechnique Separation of poly-β-hydroxyalkanoic acid from microbial biomass
JPH11511025A (ja) * 1995-08-21 1999-09-28 ザ、プロクター、エンド、ギャンブル、カンパニー バイオマスからのポリヒドロキシアルカノエートの溶媒抽出
JP2002543794A (ja) * 1999-05-12 2002-12-24 メタボリックス，インコーポレイテッド ポリヒドロキシアルカノエートを精製するための方法
JP2002262886A (ja) * 2001-03-14 2002-09-17 Inst Of Physical & Chemical Res 短鎖脂肪酸由来のモノマーからなる共重合ポリエステルを合成する植物及びポリエステルの製造方法
JP2002306190A (ja) * 2001-04-10 2002-10-22 Canon Inc 生体細胞からのポリ−３−ヒドロキシアルカン酸の分離・回収方法
WO2005085461A1 (fr) * 2004-03-04 2005-09-15 Kaneka Corporation Methode de degradation de l'acide nucleique et son utilisation
JP2005348640A (ja) * 2004-06-09 2005-12-22 Asahi Breweries Ltd Ｐｈａ精製方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ROH K.S. ET AL.: "The Effects of Sodium Bisulfite in Extraction of PHB by Hypochloride", BIOTECHNOL. TECH., vol. 9, no. 10, 1995, pages 709 - 712, XP002990865 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025134498A1 (fr) * 2023-12-18 2025-06-26 三菱瓦斯化学株式会社 Procédé de production d'agrégat d'acide polyhydroxyalcanoïque

Also Published As

Publication number Publication date
JPWO2008026619A1 (ja) 2010-01-21

Similar Documents

Publication Publication Date Title
JP5334994B2 (ja) ポリ−３−ヒドロキシアルカン酸の製造方法およびその凝集体
JPWO2004065608A1 (ja) 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
JP5651017B2 (ja) ポリ−３−ヒドロキシアルカン酸の生産方法
WO2010116681A1 (fr) Procédé de récupération de polyhydroxyalcanoate
EP1705250B1 (fr) Procede de separation, d'extraction et de purification de poly-beta-hydroxyalcanoates (pha) directement a partir d'un milieu de culture bacterien fermente
CN1685048A (zh) 3－羟基链烷酸共聚物的精制方法
CN115058461B (zh) 一种从发酵液中直接分离提纯聚羟基烷酸酯的方法
JP6864585B2 (ja) ポリヒドロキシアルカノエートの製造方法
JP2008054541A (ja) 醗酵生産された３−ヒドロキシアルカン酸共重合体の分離精製方法
WO2008026619A1 (fr) Procede de production de copolymeres d'acide 3-hydroxyalcanoique
JPWO2003091444A1 (ja) ポリ−３−ヒドロキシアルカン酸の分離方法
JPWO2005085461A1 (ja) 核酸の分解方法及びその用途
EP1550724A1 (fr) Methode de coagulation de l'acide poly-3-hydroxyalcanoique
CN116144046B (zh) 一种聚羟基脂肪酸酯凝集体的制备方法
JP7641947B2 (ja) ポリヒドロキシ酪酸系樹脂の製造方法
Salmiati et al. Recovery of polyhydroxyalkanoates (PHAs) from mixed microbial cultures by simple digestion and saponification
JP2001057895A (ja) ポリ−３−ヒドロキシアルカン酸の抽出方法
WO2022215335A1 (fr) Procédé de production d'eau recyclée
JP2009207380A (ja) ポリヒドロキシアルカン酸の精製方法

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 07806202

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2008532082

Country of ref document: JP

NENP Non-entry into the national phase

Ref country code: DE

NENP Non-entry into the national phase

Ref country code: RU

122 Ep: pct application non-entry in european phase

Ref document number: 07806202

Country of ref document: EP

Kind code of ref document: A1