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WO1998007879A1 - Procede d'obtention de polyhydroxyalcanoates et leur utilisation - Google Patents

Procede d'obtention de polyhydroxyalcanoates et leur utilisation Download PDF

Info

Publication number
WO1998007879A1
WO1998007879A1 PCT/DE1997/001772 DE9701772W WO9807879A1 WO 1998007879 A1 WO1998007879 A1 WO 1998007879A1 DE 9701772 W DE9701772 W DE 9701772W WO 9807879 A1 WO9807879 A1 WO 9807879A1
Authority
WO
WIPO (PCT)
Prior art keywords
cell
dispersion
polyhydroxyalkanoates
cell mass
aqueous phase
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/DE1997/001772
Other languages
German (de)
English (en)
Inventor
Klaus Metzner
Marion Sela
Dietmar Runkel
Torsten Sembritzki
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.)
Dow Olefinverbund GmbH
Original Assignee
Buna Sow Leuna Olefinverbund GmbH
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 Buna Sow Leuna Olefinverbund GmbH filed Critical Buna Sow Leuna Olefinverbund GmbH
Publication of WO1998007879A1 publication Critical patent/WO1998007879A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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

  • PHA polyhydroxyalkanoates
  • solution and precipitation processes are known in which the PHA is removed from the cell structure in dissolved form and separated from the residual biomass.
  • 1,2-dichloroethane in a mixture with ethanol, according to EP 024810 dichloromethane and according to US 32756 0 chloroform are used as solvents for the PHA.
  • suitable precipitants the PHA is obtained in a solid form from the PHA-containing solution separated from the cell mass.
  • polyhydroxybutyric acid is extracted from the biomass with acetic anhydride at elevated temperatures, here at 100 to 140 ° C.
  • the polyhydroxybutyric acid precipitates out of the polymer solution in a powdery state by lowering the temperature.
  • the handling of large amounts of solvents and the associated recycling and disposal costs have a disadvantageous effect. This applies in particular to chlorinated hydrocarbons as solvents.
  • these processes are often associated with severe molecular weight loss and the formation of a crystalline PHA fraction.
  • Patent application WO 95/08620 describes a recovery process for a solid product contained in a cell mass, in which the cell mass is converted into a dissolved form and is separated off together with the aqueous fermentation broth in such a way that the insoluble product, for example indigo contained in the cell mass , is in solid form.
  • the cell substances are dissolved by adding an alkali metal hydroxide solution at a treatment temperature between 40 and 100 ° C.
  • the extraction process for cell constituents can also be carried out by lysing the cell substances by means of an enzymatic treatment, in particular by using lysozyme, proteases, DNAsen, RNAsen or a combination of these.
  • patent application WO 94/24302 describes a method which uses an oxidizing medium in the presence of a complexing agent to dissolve the cell shells of the microorganisms.
  • a hydrogen peroxide at a treatment temperature between 60 and 180 ° C is used as an oxidizing agent for the production of polyhydroxybutyric acid.
  • a physical preparation of the cell material is possible, in particular through a temperature treatment in the range between 100 and 200 ° C.
  • the object of the invention is to obtain a polyhydroxyalkanoate, in particular polyhydroxybutyric acid, polyhydroxyvaleric acid or a copolymer of these with a particle size of less than 1.5 ⁇ m from an aqueous cell suspension of a microorganism that accumulates these polymers and to use them. It is necessary that the granules of the polymer embedded in the cells of the microorganisms in the amorphous state are retained as unchanged as possible in this state and that a proportion of the crystalline structure of the polymer is excluded or kept negligibly low.
  • the object is achieved by the invention presented in the claims.
  • the present method is characterized in that, without the use of PHA solvents, a dispersion with a high proportion of polyhydroxyalkanoates is formed by combining mechanical, chemical and enzymatic treatments of the bacterial biomass suspension with intermediate separation of the extracted cell components.
  • the PHA content achieved depends on the PHA content of the biomass and the number of processing steps used.
  • the cell walls are first destroyed by the action of mechanical forces on the microorganism cells in the aqueous phase.
  • Such mechanical forces can be generated in known devices by the generation of shock, friction, shear or pressure forces. are caused. It is also possible to let such forces act on the cell material in their pure form, as well as in superimposed or alternating form.
  • the bacterial biomass is subjected to mechanical cell disruption, for example in an agitator ball mill or a high-pressure homogenizer.
  • the disruption serves to destroy the solid cell envelope and release water-soluble or water-rinsable cell components. Furthermore, an enlarged surface for the attack of biological or chemical substances is created.
  • the separation between the treatment steps is carried out by separation, centrifugation or by decanter. This removes dissolved substances, fine cell fragments and impurities in the clear run, which in each case leads to a concentration of the polyhydroxyalkanoate granules.
  • the following step is followed by an alkaline treatment, for example with sodium hydroxide solution, in a pH range of 8-12. This treatment is accompanied by heating the suspension to a temperature above 70 ° C and leads to the hydrolysis of certain cell components.
  • an enzymatic purification preferably of a proteolytic character
  • a lower residual biomater content and thus a purer product high storage stability and the possibility of producing easily resuspendable dispersion powders, which represent the most stable storage form, are achieved.
  • Mixtures of cell wall-lysing enzymes in combination with pure proteases have proven successful.
  • hydrogen peroxide bleaching is carried out at a temperature between 70 ° C. to 95 ° C., followed by washing.
  • Such polymer particles obtained from cells of microorganisms by the method according to the invention are particularly suitable for the production of polymer dispersions. This can be done directly or by resuspending the PHA particles present in dried form according to the recovery process described in a liquid medium, preferably in water. It is possible to use dispersing aids, film formers, anti-foaming agents, etc. The proportion of solid phase to liquid phase depends on the intended application of the corresponding dispersion.
  • these dispersions are suitable for the production of coatings or coatings on surfaces.
  • these can be surfaces of a cellulose material, such as cardboard or paper, since in addition to the barrier properties, this ensures that these coated materials are completely biodegradable. In addition to being applied to surfaces, this effect can also be achieved by introducing the dispersion into the paper pulp.
  • a dispersion produced by the process according to the invention can be used as an auxiliary for the processing of polymers, in particular when processing PVC.
  • the nitrogen content in the polyhydroxyalkanoate has a stabilizing effect on the polymer to be processed.
  • dispersions made from them are suitable as a carrier material for the introduction of medical contrast agents and active ingredients.
  • a fermentation process in a batch process cultivates microorganisms through a corresponding growth phase with subsequent initiation of accumulation for the storage of polyhydroxybutyric acid in the cell.
  • the polymer concentration that can be achieved depends very much on the type of microorganism and the fermentation conditions. For technically relevant processes, a polymer content of more than 50% of the dry cell mass is assumed.
  • 1st step cell disruption in a high-pressure homogenizer (850 bar, 2 passages) and separation of cell fragments via a plate separator (5000 g)
  • 3rd step hydrogen peroxide bleaching with H 2 O (33%) in a ratio of 1:10 for 2 h
  • Step 4 washing and thickening using a plate separator
  • PHB content of the solid 78% nitrogen content: 0.7%
  • the target grain range is clearly achieved.
  • the indication of the nitrogen content in the product reflects the residual biomass content and serves as a measure of the product purity.
  • 3rd step hydrogen peroxide bleaching with H 2 O 2 (33%) in a ratio of 1:10 for 4 h
  • Step 4 washing and thickening using a plate separator
  • 3rd step enzyme treatment by Sfreptomyces species in connection with
  • Example 2 the possibility of producing a dispersion powder with subsequent resuspension was investigated.
  • the crude dispersion prepared in Example 2 was used for this.
  • Step 5 spray drying the dispersion to produce a dipersion powder
  • Step 6 Resuspending in water using Ultraturrax
  • the experiment was carried out on the basis of a biomass with a significantly higher starting PHB content.
  • a PHB content 76% was achieved after the fermentation process had ended.
  • the starting biomass concentration based on the dry matter was 52 g / l.
  • the fermenter broth was already concentrated here using a plate separator and brought to a dry biomass content of 137 g / l.
  • 1st step cell disruption in a high-pressure homogenizer (1000 bar, 1 passage) and separation of cell fragments via a plate separator (5000 g)
  • 2nd step Sodium hydroxide solution treatment at 75 ° C. and pH 10 for 2 h, washing over
  • 3rd step hydrogen peroxide bleaching with H 2 O 2 (33%) in a ratio of 1:10 for 2 h 80 ° C

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 un procédé permettant d'obtenir des polyhydroxyalcanoates à l'état amorphe à partir des structures cellulaires d'un microorganisme accumulant les alcanoates. Selon ledit procédé, la masse cellulaire en phase aqueuse est d'abord soumise à des forces mécaniques, puis à un traitement alcalin à des températures supérieures à 70 °C, et à une lyse. Elle est ensuite traitée avec du peroxyde d'oxygène à des températures allant de 70 à 95 °C. Les composants cellulaires non polymères sont séparés entre les différentes étapes du procédé. Les particules de polyhydroxyalcanoates ainsi obtenues peuvent être utilisées en tant que dispersion polymère pour constituer un revêtement de surface, comme agent auxiliaire de traitement pour polymères, ou bien comme matériau de support en médecine.
PCT/DE1997/001772 1996-08-20 1997-08-19 Procede d'obtention de polyhydroxyalcanoates et leur utilisation Ceased WO1998007879A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19633475.6 1996-08-20
DE1996133475 DE19633475C2 (de) 1996-08-20 1996-08-20 Verfahren zur Gewinnung von Polyhydroxyalkanoaten und deren Verwendung

Publications (1)

Publication Number Publication Date
WO1998007879A1 true WO1998007879A1 (fr) 1998-02-26

Family

ID=7803079

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/DE1997/001772 Ceased WO1998007879A1 (fr) 1996-08-20 1997-08-19 Procede d'obtention de polyhydroxyalcanoates et leur utilisation

Country Status (2)

Country Link
DE (1) DE19633475C2 (fr)
WO (1) WO1998007879A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20010009719A (ko) * 1999-07-13 2001-02-05 성재갑 미생물로부터 폴리하이드록시알카노에이트를 분리하는 방법
US6436639B1 (en) 1997-02-18 2002-08-20 Tanox, Inc. Bak promoter expression system
EP1245682A3 (fr) * 2001-03-27 2003-09-17 Canon Kabushiki Kaisha Procédé et appareil pour la production de polyhydroxyalcanoate
WO2005052175A3 (fr) * 2003-11-28 2005-06-30 Phb Ind Sa Procede de recuperation de polyhydroxialcanoates ('phas') dans une biomasse cellulaire
EP1550723A4 (fr) * 2002-09-30 2006-01-18 Kaneka Corp Methode de purification d'un copolymere d'acide 3-hydroxyalcanoique
WO2022090960A1 (fr) * 2020-10-30 2022-05-05 Biotrend - Inovação E Engenharia Em Biotecnologia, S.A. Procédé d'extraction et de purification de polyhydroxyalcanoates

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20230097635A (ko) * 2021-12-24 2023-07-03 씨제이제일제당 (주) 세포 파쇄 모니터링 방법 및 이를 이용한 폴리하이드록시알카노에이트를 생산하기 위한 방법

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0145233A2 (fr) * 1983-11-23 1985-06-19 Imperial Chemical Industries Plc Procédé de séparationpour un polymerede butyrate-3-hydroxy
WO1992022659A1 (fr) * 1991-06-11 1992-12-23 Kanebo, Ltd. Procede de production d'un ester d'acide organique polyhydroxy
WO1994024302A1 (fr) * 1993-04-14 1994-10-27 Zeneca Limited Production de matieres plastiques a partir de micro-organismes
EP0622462A1 (fr) * 1993-04-29 1994-11-02 Repsol Quimica S.A. Procédé d'extraction de polyhydroxyalkanoates à partir de bactéries halophilique les contenant
JPH0779787A (ja) * 1993-09-13 1995-03-28 Denki Kagaku Kogyo Kk ポリヒドロキシアルカノエートの分離精製法
WO1995033065A1 (fr) * 1994-06-01 1995-12-07 The Procter & Gamble Company Procede de recuperation de polyhydroalcanoates au moyen d'un fractionnement centrifuge
WO1995033064A1 (fr) * 1994-06-01 1995-12-07 The Procter & Gamble Company Procede de recuperation de polyhydroxyalcanoates en utilisant une classification par air

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2527619A1 (fr) * 1982-05-27 1983-12-02 Ici Plc Procede pour la separation d'un polymere a partir d'une solution par formation d'un gel et evaporation
DE3630778A1 (de) * 1986-09-10 1988-03-24 Neynaber Chemie Gmbh Gleitmittelsystem fuer die verarbeitung von hart-pvc
GB2244714B (en) * 1990-05-31 1993-10-06 Sanyo Chemical Ind Ltd Foamed polyurethane-forming composition,foamed polyurethane and process for making the same
DE4120582A1 (de) * 1991-06-21 1992-12-24 Hoechst Ag Bindemittelharze zur herstellung von faserverbunden
GB9223709D0 (en) * 1992-11-12 1992-12-23 Ici Plc Process for the separation of protein from microorganisms

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0145233A2 (fr) * 1983-11-23 1985-06-19 Imperial Chemical Industries Plc Procédé de séparationpour un polymerede butyrate-3-hydroxy
WO1992022659A1 (fr) * 1991-06-11 1992-12-23 Kanebo, Ltd. Procede de production d'un ester d'acide organique polyhydroxy
WO1994024302A1 (fr) * 1993-04-14 1994-10-27 Zeneca Limited Production de matieres plastiques a partir de micro-organismes
EP0622462A1 (fr) * 1993-04-29 1994-11-02 Repsol Quimica S.A. Procédé d'extraction de polyhydroxyalkanoates à partir de bactéries halophilique les contenant
JPH0779787A (ja) * 1993-09-13 1995-03-28 Denki Kagaku Kogyo Kk ポリヒドロキシアルカノエートの分離精製法
WO1995033065A1 (fr) * 1994-06-01 1995-12-07 The Procter & Gamble Company Procede de recuperation de polyhydroalcanoates au moyen d'un fractionnement centrifuge
WO1995033064A1 (fr) * 1994-06-01 1995-12-07 The Procter & Gamble Company Procede de recuperation de polyhydroxyalcanoates en utilisant une classification par air

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 123, no. 1, 3 July 1995, Columbus, Ohio, US; abstract no. 8042, YAMAMOTO, OSAMU ET AL: "Separation and purification of poly(hydroxyalkanoates) from microorganisms using surfactants" XP002051163 *

Cited By (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6436639B1 (en) 1997-02-18 2002-08-20 Tanox, Inc. Bak promoter expression system
KR20010009719A (ko) * 1999-07-13 2001-02-05 성재갑 미생물로부터 폴리하이드록시알카노에이트를 분리하는 방법
EP1245682A3 (fr) * 2001-03-27 2003-09-17 Canon Kabushiki Kaisha Procédé et appareil pour la production de polyhydroxyalcanoate
US6808907B2 (en) 2001-03-27 2004-10-26 Canon Kabushiki Kaisha Method and apparatus for producing polyhydroxyalkanoate
EP1550723A4 (fr) * 2002-09-30 2006-01-18 Kaneka Corp Methode de purification d'un copolymere d'acide 3-hydroxyalcanoique
US7435566B2 (en) 2002-09-30 2008-10-14 Kaneka Corporation Method of purifying 3-hyroxyalkanoic acid copolymer
WO2005052175A3 (fr) * 2003-11-28 2005-06-30 Phb Ind Sa Procede de recuperation de polyhydroxialcanoates ('phas') dans une biomasse cellulaire
US9045595B2 (en) 2003-11-28 2015-06-02 Phb Industrial S.A. Process for recovering polyhydroxialkanoates (“PHAs”) from cellular biomass
WO2022090960A1 (fr) * 2020-10-30 2022-05-05 Biotrend - Inovação E Engenharia Em Biotecnologia, S.A. Procédé d'extraction et de purification de polyhydroxyalcanoates

Also Published As

Publication number Publication date
DE19633475A1 (de) 1998-02-26
DE19633475C2 (de) 2002-03-14

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