EP4237467A1 - Verfahren zur extraktion und reinigung von polyhydroxyalkanoaten - Google Patents

Verfahren zur extraktion und reinigung von polyhydroxyalkanoaten

Info

Publication number
EP4237467A1
EP4237467A1 EP21805641.4A EP21805641A EP4237467A1 EP 4237467 A1 EP4237467 A1 EP 4237467A1 EP 21805641 A EP21805641 A EP 21805641A EP 4237467 A1 EP4237467 A1 EP 4237467A1
Authority
EP
European Patent Office
Prior art keywords
pha
recovered
biomass
extraction
process according
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.)
Pending
Application number
EP21805641.4A
Other languages
English (en)
French (fr)
Inventor
Bruno Sommer Ferreira
João Manuel BEIRÃO TORNEIRO CAVALHEIRO
Hugo FERRÃO DIAS DE ALMEIDA
Joshua ANJOS
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.)
Biotrend Inovacao E Engenharia Em Biotecnologia SA
Original Assignee
Biotrend Inovacao E Engenharia Em Biotecnologia SA
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 Biotrend Inovacao E Engenharia Em Biotecnologia SA filed Critical Biotrend Inovacao E Engenharia Em Biotecnologia SA
Publication of EP4237467A1 publication Critical patent/EP4237467A1/de
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
    • C08G63/88—Post-polymerisation treatment
    • C08G63/90—Purification; Drying
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
    • C08G63/00—Macromolecular compounds obtained by reactions forming a carboxylic ester link in the main chain of the macromolecule
    • C08G63/91—Polymers modified by chemical after-treatment
    • C08G63/912—Polymers modified by chemical after-treatment derived from hydroxycarboxylic acids
    • 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 process for extraction and puri fication of polyhydroxyalkanoates ( PHAs ) from PHA- containing material obtained from di f ferent sources .
  • Biodegradable natural polymers extracted or derived from renewable resources are steadily replacing the crude oil-based polymers .
  • Three families of biopolymers are usually researched and used : polymers directly extracted from biomass (biomass can comprise polysaccharides , proteins and various lipids ) , biomass-derived monomers treated with classical chemical and manufacturing routes to obtain biodegradable and/or renewable polymers (polylactate , bio-polyethylene ) , and polymers produced by natural or genetically modi fied micro-organisms (polyhydroxyalkanoates , PHAs ) .
  • PHAs have attracted special interest because they comprise a group of naturally derived polyesters synthesi zed by a wide range of microorganisms as intracellular carbon and energy materials . They are usually accumulated within cells when growth is l imited by nutrients such as nitrogen, oxygen, phosphorous and other essential elements , while in the presence of excess carbon . Instead of being consumed for the cellular growth, the excess of carbon is taken into the cells and stored in the form of PHA granules .
  • PHAs production at large scale from microorganisms usually involves fermentation, isolation and puri fication processes , which imply higher production costs .
  • manufacturers in Brazil and China have used sugarcane bagasse and cornstarch as a renewable resource to produce PHA inexpensively .
  • pioneering and significant research ef forts on the use of mixed microbial cultures from wastes as a cheaper alternative to pure microbial culture has been carried out , enabling the use of renewable carbon which does not compete with food or feed applications .
  • Such consortia of PHA-producing microorganisms can adapt to changes in the substrate and enable the use of cheap mixed substrates such as low cost agricultural or industrial waste feedstock and even municipal wastes .
  • PHA polyhydroxybutyrate
  • PHB polyhydroxybutyrate
  • PHBV poly ( p-hydroxybutyrate-co-valerate )
  • PHB, PHBV and other short-chain length co-polymers are important biodegradable plastics that are becoming of signi ficant interest in food packaging applications .
  • the downstream processing for PHAs recovery and puri fication from microbial biomass plays a vital role in the PHAs manufacturing process . It is a maj or contributor to the product production cost , it defines the material speci fications and quality, and may have a significant impact on the environmental sustainability of the product and of its production process .
  • the PHA puri fication step is often recogni zed as the most critical when assessing the process feasibility and sustainability .
  • the traditional puri fication processes relying on organic solvents , including halogenated, are being banned from the industrial setting, while new hopes of using microbes that spontaneously release the PHA to the medium ( ex . upon osmotic shock) have not yet been realized due to their low productivity .
  • the steps of extraction and puri fication of PHAs from producing biomass are commonly regarded as among those that contribute the most to the final price of the polymer . This is more so for the extraction of PHAs from biomass used in the treatment of municipal wastes , due to the complex matrix of materials that can adsorb the biomass and the heterogeneous nature of the mixed culture . Further, some applications of the puri fied polymer may require obtaining stringent speci fication, whereas less demanding applications may exist that can eventually use non- fully puri fied material s while complying with the required functional features and relevant regulatory framework .
  • EP 1705250 Al entitled "A method for separating, extracting and puri fying poly-beta-hydroxyalkanoates ( PHAs ) directly from bacterial fermented broth” discloses a method for directly separating and purifying polyhydroxyal kanoates in cells from fermentation liquid and it relates to the field of post-treatment technology of biological engineering .
  • the method includes the steps of : pretreating fermentation liquid with physical method for breaking cell wall ; adj usting the pH value of the pretreated fermentation liquid to alkaline ; adding an anionic surfactant ; reacting under agitation; separating and extracting the coagulated precipitate from the reaction liquid; washing and drying .
  • the invention was so far only validated on pure cultures grown on standard industrial fermentation media, which entrains much less contaminants than the challenging conditions of the streams emerging from the mixed culture fermentations .
  • EP 2606080 Bl entitled “Method for recovery of stabili zed polyhydroxyalkanoates from biomass that has been used to treat organic waste” relates to a method of increasing the chemical and/or thermal stability of PHA in biomass where the biomass is contained within mixed liquor, and wherein the mixed liquor is treated by a combination of removing water from the mixed liquor and pH adj ustment of the mixed liquor or maintenance of the pH of the mixed liquor within a selected pH range , and wherein the method includes reducing the pH of the mixed liquor below 6 , or maintaining the pH o f the mixed liquor below 6 for a selected period of time , and wherein the pH adj ustment of the mixed liquor to below 6 or the maintenance of the pH of the mixed liquor below 6 gives rise to an increase in chemical and/or thermal stability of the PHA in the biomass .
  • the PHA is extracted with a non-chlorinated organic solvent at temperatures above 100 ° C .
  • the process was designed to provide high purity PHAs with enhanced thermal stability .
  • such process relies on the use of organic solvents processed at high temperature ( typically at more than 100 ° C ) , which add on cost and would require equipment and safety features which may not be usually employed in the plants producing the PHA- containing biomass , namely wastewater treatment plants .
  • EP 3287526 Bl entitled "Method of manufacturing microbially produced plastic and microbially produced plastic” , relates to a method of manufacturing a microbially produced plastic comprising a step of applying heat treatment to fat containing hydrogen peroxide and a step of culturing microbes in a culture liquid containing the fat that has been subj ected to the heat treatment .
  • the extraction process should use standard and readily available equipment , preferably equipment that can be routinely found on-site and not requiring special training for the plant operators . This precludes the use of processes using organic solvents .
  • the present invention provides a process for extraction and purification of polyhydroxyalkanoates (PHA) characterized by reacting PHA-containing material with H2O2 and adding an alkali up to a pH value equal to 9 or higher.
  • PHA polyhydroxyalkanoates
  • the reaction is carried out at a pH value above 11, more preferably above 11,4 and most preferably above 11,5.
  • the ratio of H2O2 to PHA-containing material in the process of the invention is within a range of 20 mL H2O2 30 vol/g of biomass (dry weight) to 0.1 mL H2O2 30vol/g of biomass (dry weight) .
  • the alkali can be added simultaneously with or sequentially to H2O2.
  • the alkali of the reaction is selected from a group consisting of NaOH, KOH and Ca(OH)2, with NaOH being preferred.
  • the PHA-containing material is a mixed microbial culture or a pure culture containing intracellular PHAs .
  • the process of the invention further comprises a pre-treatment step which may be chemical, mechanical, electric or enzymatic.
  • the present invention provides a process for extraction and puri fication of polyhydroxyalkanoates ( PHA) characteri zed by reacting PHA- containing material with H2O2 and adding an alkali up to a pH value equal to 9 or higher .
  • PHA polyhydroxyalkanoates
  • the process of the invention comprises the steps of : a ) adding a PHA-containing material to a reaction vessel b ) adj usting the pH to a value of 9 or higher by adding an alkali to the PHA-containing material under stirring; c ) adding H2O2 and alkali over 30 to 90 minutes under stirring; d) submitting the mixture to a digestion period by stirring for 2 , 5 to 36 hours followed by separation, washing and drying steps .
  • PHA-containing material means any PHA-containing microbial biomass or streams containing or derived from PHA-containing microbial biomass , including pre-treated microbial biomass .
  • PHA-containing microbial biomass means any microbial biomass containing intracellular PHA, including microbial biomass consisting on or derived from pure microbial cultures , microbial biomass consisting on or derived from mixed microbial cultures , microbial biomass consisting on or derived from naturally occurring microorganisms , microbial biomass consisting on or derived from genetically modified microbial cultures, or any combination thereof.
  • the reaction is carried out at a pH value above 11, more preferably above 11,4 and most preferably above 11,5.
  • the ratio of H2O2 to PHA-containing material in the process of the invention is within a range of 20 mL H2O2 30 vol/g of biomass (dry weight) to 0.1 mL H2O2 30vol/g of biomass (dry weight) .
  • the alkali can be added simultaneously with or sequentially to H2O2.
  • the alkali of the reaction is selected from a group consisting of NaOH, KOH and Ca(OH)2, with NaOH being preferred .
  • PHA-containing material is a mixed microbial culture or a pure culture containing intracellular PHAs .
  • the process of the invention may further comprise a pre-treatment step of which may be chemical, mechanical, electric or enzymatic.
  • the chemical pre-treatment comprises the pre-treatment of the microbial biomass with a surfactant and/or with a bleaching agent and/or with an alkali.
  • the surfactant can be any surfactant which interferes with the integrity of the cellular membrane of microbial cells such as, for example, sodium dodecyl sulphate and similar.
  • the bleaching agent is selected from sodium hypochlorite or hydrogen peroxide and the alkali is selected from the group consisting of NaOH, KOH and
  • the enzymatic pre-treatment i s any treatment using enzymes involved in hydrolysing components of the microbial cell walls , either isolated or in combination, including, without limitation, proteases , lipases , carbohydrases or the like .
  • the mechanical pre-treatment is selected from any treatment allowing cell disruption, such as sonication, high pressure homogenisation, milling, and the like .
  • the electric pre-treatment is selected from any treatment allowing cell disruption, such as pulsed electric f ield processing, and the like .
  • the process of the present invention was found to be suitable to obtain a viable product and contrary to known processes , it is easy to be controlled without violent reactions which generally occurs due to high temperatures and/or the release of gases .
  • the present process is ef fective for extraction of PHAs from very diverse microbial biomass , including pure cultures , and mixed cultures obtained from the conversion of di f ferent wastes .
  • the process of the invention exhibits further advantages : Processing of both cleaner biomasses ( ex . pure microbial cultures obtained from the fermentation o f refined raw materials ) and crude biomasses ( ex . pure microbial cultures obtained from the fermentation of industrial sidestreams or mixed cultures obtained from the conversion of municipal or agro-industrial wastes ) ;
  • the digestion reaction can be performed without temperature control ;
  • the conditions can be set in order to minimise the ef fect of the digestion reaction on the polymer molecular weight ;
  • the polymer allows obtaining white formulations and white plastic parts (very important to allow the incorporation of coloured pigments in the plastics ) , while the solutions currently on the market originate beige products ;
  • the final product does not contain a chlorinated smell ; by using the processes of prior art , a typical chlorine smell was detected on the resulting polymer powder even after multiple washing steps and this is very relevant for processing, final use and product stability, since when residual chlorinated compounds remained on the f inal product , chlorine vapors were released when processing the polymer and corrosion issues on the processing equipment could arise ;
  • PHA-containing microbial biomass was produced from the fermentation of fruit pulp, an industrial by-product of the j uice industry, using mixed microbial cultures .
  • a first stage of the process consists of acidogenic fermentation in which the available feedstock is converted into volatile fatty acids , such as , but not limited to , acetic acid, lactic acid, propionic acid, butyric acid, valeric acid, caproic acid and also ethanol .
  • volatile fatty acids such as , but not limited to , acetic acid, lactic acid, propionic acid, butyric acid, valeric acid, caproic acid and also ethanol .
  • a cyclic regime of carbon excess and carbon limitation was established to enhance PHA storage capacity ( generic information on PHA production using mixed microbial cultures can be obtained, for example , in Serafim et al . , 2008 and speci fic information of the system used for PHA production using fermentation of fruit pulp in Melendez-Rodriguez et al
  • the PHA recovery yield decreased with decreasing amounts of sodium hypochlorite ( 100% , 58 % and 32 % when 400 g, 300 g and 200 g were used, respectively) , suggesting that the reduction of the quantities of sodium hypochlorite did not allow the ef fective digestion of the non-PHA microbial biomas s components .
  • a small amount was placed on a glass slide which was slowly heated in a heating plate up to a maximum of 180 ° C . Most of the material recovered us ing 200 g of bleach did not melt and became brown upon heating .
  • the milder conditions 1- 1 using a lower concentration of SDS and remaining reagents , resulted in a very low PHA recovery and the recovered material did not melt properly, generating a mostly brown residue when heating .
  • the remaining conditions resulted in higher yield and the recovered material melted upon heating without generating brown or even yellowish residues , a strong indication that the recovered material had a good purity .
  • the total consumption is still very high ( 5- 10 times the amount of PHA-containing microbial biomas s on dry weight ) and the final product still retained a strong chlorine smell .
  • experiment 3-1 an unexpectedly fast expansion of the reaction medium occurred due to vigorous gas release and which was very difficult to contain even through the addition of silicone-based anti-foaming agent (Simethicone 30% emulsion, from Dow-Corning) .
  • experiment 3-2 the volume expansion also occurred vigorously, but could be managed with the addition of anti- foaming agent .
  • experiment 3-3 only a slight volume expansion occurred, and the addition of antifoaming agent was not required .
  • a signi ficant amount of the recovered material did not melt , no signi ficant browning was observed upon melting . This indicates that most organic non-PHA microbial biomass components have been digested, but the recovered polymer is contaminated by a signi ficant amount of inorganics which were not fully removed in the washing process .
  • Example 1A - PHA extraction Controlled use of sodium hydroxide and hydrogen peroxide - pH 12 . 5
  • Example IB - PHA extraction Controlled use of sodium hydroxide and hydrogen peroxide - pH 9.0
  • Example 1C - PHA extraction Controlled use of sodium hydroxide and hydrogen peroxide - pH 13.0
  • an initial amount of sodium hydroxide is added in order to bring the pH of the reaction medium at around 13.5.
  • hydrogen peroxide and sodium hydroxide are slowly added over 90 minutes without temperature control.
  • a total of 15 mL of 30 volume strength hydrogen peroxide and 20 mL of sodium hydroxide (5 N solution) are added, after which the mixture is stirred for 2.5 hours. After this digestion period, the mixture is centrifuged and the solids are recovered.
  • the thus recovered solids are resuspended in water, using the same volume as the digestion reaction volume, and the wash water discarded. Four additional wash cycles are carried out.
  • the washed and recovered solids are oven dried at 60°C until constant weight and the recovered material is ground to a powder in a mortar.
  • the PHA recovery yield is 67% and upon heating a mostly transparent melt was obtained at 175°C with rare yellowish spots, indicating that the material is compatible with many of the envisaged applications.
  • Example 2 - PHA extraction Combined use of surfactant, sodium hypochlorite, and controlled use of sodium hydroxide and hydrogen peroxide
  • PHB-containing microbial biomass was produced from the aerobic fermentation of sugars using a Burkholderla saccharl strain (for details see Cesario et al . , 2014 ) .
  • the PHB- containing microbial biomass with a dry weight of 35% on total wet microbial biomass and 49% of PHB on dry weight , was diluted with water to a 20% dry cell weight and mixed with SDS and a solution of NaOH 5N, both to 5% of the total volume .
  • the lower amount of chemicals used is due to the nature of the treated microbial biomass , which is more homogenous (a pure microbial culture instead of mixed microbial culture ) and which was fermented on a cleaner culture medium ( a defined fermentation medium instead of industrial organic residues ) .
  • domestic bleach was added to 20% of the volume and mixing continued for 60 minutes After this digestion period, the solids were recovered by centri fugation . The thus recovered solids were resuspended in water, using the same volume as the digestion reaction volume , and the wash water discarded . The washed and recovered solids were oven dried at 60 ° C until constant weight and the recovered material was ground to a powder in a mortar .
  • a recovery yield of 91% was obtained, and the purity as assessed by the melting behaviour of the recovered powder was excellent, with a homogenous melt, without any brown or yellowish spots.
  • the thus obtained polymer was used in a formulation to produce a melt which was then extruded (Haake Rheocord 90 single screw extruder) , and successfully produced a filament of 1.810.1 mm with adequate quality for 3D printing tests. Further, the polymer originated a white extruded filament that could be used to produce a white plastic part.
  • Example 3A 4.5 kg of PHB-containing microbial biomass described in Example 3A, was adjusted to a pH of 11.9 with NaOH 5N. Subsequently, 30 volume strength hydrogen peroxide were slowly added at a rate of 40 mL/h. The pH of the reactor was allowed to decrease as hydrogen peroxide was added. When the pH reached 11.2, the automated addition of NaOH was implemented in order to control the pH at that setpoint. A total of 320 mL of hydrogen peroxide and 75 mL of NaOH were added and 1.1 kg of wet pellet was recovered. A second digestion step was carried out using the same conditions as for the first digestion step, with a total addition of 125 mL hydrogen peroxide and 90 mL of NaOH.
  • the controlled addition of the chemicals enabled to avoid extensive foam formation and also to control the temperature of the reaction, which did not exceed 37°C.
  • the thus recovered solids were resuspended in water, using the same volume as the digestion reaction volume, and the wash water discarded. Two additional washing steps were carried out. The washed and recovered solids were oven dried at 60°C until constant weight and the recovered material was ground to a powder in a mortar, resulting in a white material with good melting properties.
  • Table 5 Effect of the extraction on hydroxybutyrate (HB) and hydroxyvalerate (HV)
  • the parameters related to the concentration and ranges of the reagents as well the duration of the digestion/ reaction are interrelated .
  • the skilled person taking into account the teachings of the present invention is able to choose the proper parameters in order to control the purity and the molecular weight of the extracted polymer .
  • the temperature of the reaction is also selected upon the digestion of speci fic microbial biomasses and targeting speci fic polymer speci fications .

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Medicinal Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
EP21805641.4A 2020-10-30 2021-10-27 Verfahren zur extraktion und reinigung von polyhydroxyalkanoaten Pending EP4237467A1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
PT11686520 2020-10-30
PCT/IB2021/059931 WO2022090960A1 (en) 2020-10-30 2021-10-27 Process for extraction and purification of polyhydroxyalkanoates

Publications (1)

Publication Number Publication Date
EP4237467A1 true EP4237467A1 (de) 2023-09-06

Family

ID=78536451

Family Applications (1)

Application Number Title Priority Date Filing Date
EP21805641.4A Pending EP4237467A1 (de) 2020-10-30 2021-10-27 Verfahren zur extraktion und reinigung von polyhydroxyalkanoaten

Country Status (2)

Country Link
EP (1) EP4237467A1 (de)
WO (1) WO2022090960A1 (de)

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB9307674D0 (en) * 1993-04-14 1993-06-02 Zeneca Ltd Production of plastics materials from microorganisms
DE19633475C2 (de) * 1996-08-20 2002-03-14 Buna Sow Leuna Olefinverb Gmbh Verfahren zur Gewinnung von Polyhydroxyalkanoaten und deren Verwendung
WO2004029266A1 (ja) * 2002-09-30 2004-04-08 Kaneka Corporation ３−ヒドロキシアルカン酸共重合体の精製方法
US7582456B2 (en) 2003-12-19 2009-09-01 Tianan Biologic Material Co., Ltd. Ningbo Method for separating, extracting and purifying poly-β-hydroxyalkanoates (PHAs) directly from bacterial fermentation broth
JP5784725B2 (ja) 2010-08-18 2015-09-24 ヴェオリア・ウォーター・ソリューションズ・アンド・テクノロジーズ・サポート 有機廃棄物を処理するために用いられたバイオマスからの安定化ポリヒドロキシアルカノエートの回収方法
ITMI20131276A1 (it) * 2013-07-30 2015-01-31 Bio On S R L Processo per recuperare e purificare poliidrossialcanoati da una coltura cellulare
EP3287526B1 (de) 2015-04-24 2019-12-25 Kaneka Corporation Verfahren zur herstellung eines mikrobiell hergestellten kunststoffes und mikrobiell hergestellter kunststoff

Also Published As

Publication number Publication date
WO2022090960A1 (en) 2022-05-05

Similar Documents

Publication Publication Date Title
Mannina et al. Recovery of polyhydroxyalkanoates (PHAs) from wastewater: A review
Kosseva et al. Trends in the biomanufacture of polyhydroxyalkanoates with focus on downstream processing
Singhvi et al. Biomass to biodegradable polymer (PLA)
Ong et al. An integrative study on biologically recovered polyhydroxyalkanoates (PHAs) and simultaneous assessment of gut microbiome in yellow mealworm
WO2010116681A1 (ja) ポリヒドロキシアルカノエートの回収方法
Guleria et al. Polyhydroxyalkanoates production from domestic waste feedstock: A sustainable approach towards bio-economy
Arumugam et al. Bioconversion of Calophyllum inophyllum oilcake for intensification of rhamnolipid and polyhydroxyalkanoates co-production by Enterobacter aerogenes
US7582456B2 (en) Method for separating, extracting and purifying poly-β-hydroxyalkanoates (PHAs) directly from bacterial fermentation broth
Akhlaq et al. Polyhydroxybutyrate biosynthesis from different waste materials, degradation, and analytic methods: A short review
Pan et al. Biosynthesis of polyhydroxyalkanoate from food waste oil by Pseudomonas alcaligenes with simultaneous energy recovery from fermentation wastewater
US20130164800A1 (en) Production of Polyhydroxyalkanoates
Haque et al. Advancements and current challenges in the sustainable downstream processing of bacterial polyhydroxyalkanoates
EP3894573A1 (de) Herstellung von polyhydroxyalkanoaten aus zellstoff- und papierabfallströmen
JP5784725B2 (ja) 有機廃棄物を処理するために用いられたバイオマスからの安定化ポリヒドロキシアルカノエートの回収方法
Muhorakeye et al. Valorization of pretreated waste activated sludge to organic acids and biopolymer
CN107840994A (zh) 一种可完全降解淀粉基生物塑料膜的制备方法
EP3106513A1 (de) Verfahren zur zersetzung von polyhydroxyalkansäure, herstellung und mikroorganismuszubereitung
WO2020037394A1 (en) Biodegradable polymer composition and method of producing the same
Amabile et al. An innovative and sustainable process for producing poly (3-hydroxybutyrate-co-3-hydroxyvalerate): simulating volatile fatty acid role and biodegradability
Kumar et al. Commercialization potential of agro-based polyhydroxyalkanoates biorefinery: a technical perspective on advances and critical barriers
EP4237467A1 (de) Verfahren zur extraktion und reinigung von polyhydroxyalkanoaten
Vecchiato et al. Insights into the economical production of polyhydroxyalkanoates
Alvarado-Cordero et al. Production of polyhydroxybutyrate (PHB) by bacillus megaterium DSM 32 from residual glycerol of the bioenergy industry
US20060183205A1 (en) Method for controlling molecular weight and distribution of biopolymers
Lagunes Synthesis of polyhydroxyalkanoates using orange juicing waste as feedstock

Legal Events

Date Code Title Description
STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: UNKNOWN

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE

PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE

17P Request for examination filed

Effective date: 20230518

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

DAV Request for validation of the european patent (deleted)
DAX Request for extension of the european patent (deleted)