TEPZZ_78_7 

_B T
(19)

(11) EP 1 781 721 B2

(12) NEW EUROPEAN PATENT SPECIFICATION
After opposition procedure

(45) Date of publication and mention (51) Int Cl.:

of the opposition decision: C08G 63/89 (2006.01)
26.12.2012 Bulletin 2012/52
(86) International application number:
(45) Mention of the grant of the patent: PCT/US2005/023221
11.03.2009 Bulletin 2009/11
(87) International publication number:
(21) Application number: 05763293.7 WO 2006/004895 (12.01.2006 Gazette 2006/02)

(22) Date of filing: 29.06.2005

(54) IMPROVED PROCESS FOR THE SOLVENT-BASED EXTRACTION OF

POLYHYDROXYALKANOATES FROM BIOMASS
VERBESSERTES VERFAHREN ZUR LÖSUNGSMITTELBASIERETEN EXTRAKTION VON
POLYHYDROXYALKANOATEN AUS BIOMASSE
PROCESSUS AMELIORE D’EXTRACTION A BASE DE SOLVANT DE
POLYHYDROXYALKANOATES A PARTIR DE BIOMASSE

(84) Designated Contracting States: • WELLING, Stanley, James

AT BE BG CH CY CZ DE DK EE ES FI FR GB GR Liberty Township, Ohio 45011 (US)
HU IE IS IT LI LT LU MC NL PL PT RO SE SI SK TR
(74) Representative: Vossius & Partner
(30) Priority: 29.06.2004 US 583773 P P.O. Box 86 07 67
81634 München (DE)
(43) Date of publication of application:
09.05.2007 Bulletin 2007/19 (56) References cited:
WO-A-97/07230 WO-A-2004/013204
(73) Proprietor: Kaneka Corporation WO-A1-97/07229 DE-C1- 19 955 381
Osaka-shi, US-A- 4 140 741 US-A- 4 324 907
Osaka 530-8288 (JP) US-A- 4 562 245 US-A- 5 821 299
US-A- 5 894 062 US-A- 5 942 597
(72) Inventors: US-A- 6 087 471 US-A1- 2004 053 381
• NARASIMHAN, Karunakaran US-B1- 6 340 580
West Chester, Ohio 45069 (US)
• CEARLEY, Angella, C. • DATABASE WPI Section Ch, Week 199740
Hamilton, Ohio 45015 (US) Derwent Publications Ltd., London, GB; Class
• GIBSON, Michael, Steven A23, AN 1997-429191 XP002352433 -& JP 09
Cincinnati, Ohio 45244 (US) 191893 A (MEIJI SEIKA KAISHA LTD) 29 July 1997
(1997-07-29)
EP 1 781 721 B2

Printed by Jouve, 75001 PARIS (FR)

 1 EP 1 781 721 B2 2

Description [0006] Numerous solvent-based and other types of ex-

traction techniques are known in the art for extracting
FIELD OF THE INVENTION PHAs from bacteria and plants (biomass). Solvent-based
systems (including those utilizing acetone, ketones,
[0001] This invention relates to an improved process 5 alone and in combination with other solvents), mechan-
for the extraction of specific components from other bio- ical systems, and combinations thereof may be used for
mass components. More specifically, this invention re- extracting PHA. However, known solvent-based systems
lates to an improved process for the extraction of poly- are often inefficient and may be difficult to implement with
hydroxyalkanoates from a biological system, such as a the physical characteristics of certain PHAs (problems
plant or a bacterium, by performing the extraction with a 10 with gelling, degradation, etc.) More popular are two-sol-
solvent. vent systems, but these two-solvent systems are often
expensive due to the duplicated cost of solvent and may
BACKGROUND OF THE INVENTION also create additional recovery steps when seeking to
recover/reuse both solvents.
[0002] Plastics such as polyesters are typically pro- 15 [0007] Therefore, there is a need for a more efficient
duced from petrochemical sources by well-known syn- and cost-saving process for extracting the PHA materials
thetic means. These petrochemical-based polymers take from biomass. Such a process would preferably involve
centuries to degrade after disposal. Concern over plastic recyclable solvents that are preferably environmentally
waste accumulation in landfills has resulted in the recent friendly. In addition, such a process is preferably suitable
movement toward using biodegradable polymers in- 20 to large-scale, continuous production of PHA materials.
stead.
[0003] Synthetic biodegradable polymers, also com- SUMMARY OF THE INVENTION
monly referred to as "bioplastics," have not enjoyed great
success in the marketplace due to their high production [0008] The inventors have surprisingly discovered a
cost. However, advances in biotechnology have led to 25 process for extracting PHA polymers from biomass con-
less expensive means of production. Specifically, biode- taining the PHA polymer with improved efficiency and
gradable aliphatic copolyesters are now often produced reduced cost.
by large-scale bacterial fermentation. Collectively termed [0009] The present invention therefore relates to an
polyhydroxyalkanoates or "PHAs", these polymers may improved process for extracting polyhydroxyalkanoate
be synthesized in the bodies of natural or recombinant 30 from a biomass containing the polyhydroxyalkanoate
bacteria fed with glucose in a fermentation plant. Like comprising the steps of:
their petrochemical precursors, the structural, and in turn
mechanical, properties of PHAs may be customized to a) combining the biomass containing the polyhy-
fit the specifications of the desired end product. However, droxyalkanoate with acetone as a solvent to form a
unlike their petrochemical precursors, PHAs degrade 35 biomass liquor wherein the biomass liquor compris-
both aerobically and anaerobically. es less than about 25% water;
[0004] PHAs are enormously versatile, and as many b) mixing the biomass liquor for from about 10 to
as 100 different PHA structures have been identified. about 300, alternatively from about 10 to about 240
PHA structures may vary in two ways. First, PHAs may minutes, at a temperature in the range of from about
vary according to the structure of the R-pendant groups, 40 70° C to about 120 °C;
which form the side chain of hydroxyalkanoic acid not c) separating the polyhydroxyalkanoate from the bi-
contributing to the PHA carbon backbone. Second, PHAs omass liquor to form a PHA-enriched liquor, wherein
may vary according to the number and types of units from the separating occurs at a temperature of at least
which they are derived. For example, PHAs may be about 40°C;
homopolymers, copolymers, and terpolymers. These 45 d) mixing the PHA-enriched liquor with water to form
variations in PHA structure are responsible for the vari- precipitated polyhydroxyalkanoate and an impure
ations in their physical characteristics. These physical solvent liquor, wherein the water is mixed with the
characteristics allow PHAs to be used for a number of PHA-enriched liquor in the ratio of from at least about
products which may be commercially valuable. 3 parts water to one part polyhydroxyalkanoate; and
[0005] However, in order to have any type of commer- 50 e) recovering the precipitated polyhydroxyalkanoate
cially marketable PHA bioplastic product, there is a need from the impure solvent liquor.
for identifying microbial organisms that are capable of
producing significant quantities of desirable PHA and to [0010] The present invention further relates to the
identify an efficient process for separating such PHAs above process wherein the solvent is acetone.
from the residual biomass. Improved learnings on the 55 [0011] The present invention further relates to the
biology of PHA biosynthetic pathways has allowed for above processes wherein the biomass liquor comprises
the use of microbial organisms to produce significant the solvent and the polyhydroxyalkanoate in a ratio of
quantities of PHA. from at least about 5 to about 30 parts solvent to about

2
 3 EP 1 781 721 B2 4

one part polyhydroxyalkanoate. liquor and water are mixed together by a means selected
[0012] The present invention further relates to the from propeller, turbine, high shear, and mixtures thereof.
above processes wherein the biomass liquor comprises [0027] The present invention further relates to the
the solvent and the polyhydroxyalkanoate in a ratio of above processes wherein in step (d) the PHA-enriched
from about 15 parts solvent to about 1 part polyhydroxy- 5 liquor and water are mixed together by a means selected
alkanoate. from high shear, layers of water coated sheets or moving
[0013] The present invention further relates to the belts and mixtures thereof.
above processes wherein during the step of mixing the [0028] The present invention further relates to the
biomass liquor, the temperature is from about 70°C to above processes wherein in the step of mixing the PHA-
about 100°C. 10 enriched liquor with water to form precipitated polyhy-
[0014] The present invention further relates to the droxyalkanoate and an impure solvent liquor, the PHA
above processes wherein during the step of mixing the enriched liquor is cooled to a temperature of from about
biomass liquor the mixing is performed by using mixing 20°C to about 45°C.
means selected from propellers, turbines, screw convey- [0029] The present invention further relates to the
or, and mixtures thereof. 15 above processes wherein in step (e), the precipitated pol-
[0015] The present invention further relates to the yhydroxyalkanoate recovery from the impure solvent liq-
above processes wherein the mixing means has a Pow- uor is by filtration and/or centrifugation to produce recov-
er/Volume ratio of from about 0.001 KW/m 3 to about 100 ered precipitated polyhydroxyalkanoate and remainder
KW/m3. impure solvent liquor.
[0016] The present invention further relates to the 20 [0030] The present invention further relates to the
above processes wherein the process is a continuous above processes wherein after recovery by filtration, the
process wherein during the step of mixing the biomass recovered precipitated polyhydroxyalkanoate is
liquor, the mixing is performed by using a plug flow con- squeezed and/or pressurized to remove remainder im-
cept with a screw conveyor. pure solvent liquor water.
[0017] The present invention further relates to the 25 [0031] The present invention further relates to the
above processes wherein the mixing of the biomass liq- above processes wherein in step (e), the recovered pre-
uor is conducted for a period of time selected from about cipitated polyhydroxyalkanoate is then washed with a sol-
30 to about 300 minutes. vent selected from acetone, methyl ethyl ketone, and
[0018] The present invention further relates to the mixtures thereof.
above processes wherein the biomass liquor comprises 30 [0032] The present invention further relates to the
less than 15% water. above processes wherein the process further comprises
[0019] The present invention further relates to the drying the recovered precipitated polyhydroxyalkanoate.
above processes wherein the biomass liquor comprises [0033] The present invention further relates to the
no measurable quantity of water. above processes wherein the process further comprises
[0020] The present invention further relates to the 35 recycling the wash solvent to extraction directly if the wa-
above processes wherein the separating the polyhy- ter content in the recycle wash solvent is < 15% and re-
droxyalkanoate from the biomass liquor is by filtration covering and recycling the remainder impure solvent liq-
and/or centrifugation. uor.
[0021] The present invention further relates to the [0034] The present invention further relates to an im-
above processes wherein the separating of the polyhy- 40 proved process for extracting polyhydroxyalkanoate from
droxyalkanoate from the biomass liquor is by hot filtration a biomass containing the polyhydroxyalkanoate compris-
and/or hot centrifugation. ing the steps of:
[0022] The present invention further relates to the
above processes wherein the separating of the polyhy- a) combining the biomass containing the polyhy-
droxyalkanoate from the biomass liquor occurs at a tem- 45 droxyalkanoate with acetone as a solvent to form a
perature of at least about 70°C. biomass liquor wherein the biomass liquor compris-
[0023] The present invention further relates to the es less than about 5% water;
above processes wherein the water is mixed with the b) mixing the biomass liquor for from about 30 to
PHA-enriched liquor in the ratio of from about 6 to about about 240 minutes at a temperature in the range of
8 parts water to about 1 part polyhydroxyalkanoate. 50 from about 70°C to about 120°C;
[0024] The present invention further relates to the c) separating the polyhydroxyalkanoate from the bi-
above processes wherein in step (d) the water is added omass liquor to form a PHA-enriched liquor, wherein
to the PHA-enriched liquor. the separating occurs at a temperature of at least
[0025] The present invention further relates to the about 50°C;
above processes wherein in step (d) the PHA-enriched 55 d) mixing the PHA-enriched liquor with water to form
liquor is added to the water. precipitated polyhydroxyalkanoate and an impure
[0026] The present invention further relates to the solvent liquor, wherein the water mixed with the
above processes wherein in step (d) the PHA-enriched PHA-enriched liquor in the ratio of from about 5 parts

3
 5 EP 1 781 721 B2 6

to about 30 parts water to about one part polyhy- bleaches to enhance color and odor.
droxyalkanoate; and
e) recovering the precipitated polyhydroxyalkanoate BRIEF DESCRIPTION OF THE DRAWINGS
from the impure solvent liquor; wherein the polyhy-
droxyalkanoate comprises at least two randomly re- 5 [0036]
peating monomer units, wherein the first randomly
repeating monomer unit has the structure: FIG. 1 sets forth a schematic of an embodiment of
the features detailed herein, in block form, compris-
ing the mixing, separating, precipitating, drying and
10 recovery steps.

DETAILED DESCRIPTION OF THE INVENTION

[0037] While the specification concludes with the

15 claims particularly pointing out and distinctly claiming the
and the second or higher randomly repeating mon- invention, it is believed that the present invention will be
omer unit has the structure: better understood from the following description.
[0038] All documents cited in the Detailed Description
of the Invention are, , in relevant part, incorporated herein
20 by reference; the citation of any document is not to be
construed as an admission that it is prior art with respect
to the present invention.
[0039] All percentages and ratios used herein are by
weight of the total composition and all measurements are
25 made at 25°C, unless otherwise designated. ’
wherein R is a C2 to C7 alkyl or a mixture thereof; wherein [0040] "Comprising" means that other steps and other
from about 75 mol% to about 99 mol% of the randomly ingredients, which do not affect the end result, can be
repeating monomer units have the structure of the first added. This term encompasses the terms "consisting of"
randomly repeating monomer unit and from about 1 and "consisting essentially of".
mol% to about 25 mol% of the randomly repeating mon- 30 [0041] Several processes by which PHAs may be ex-
omer units have the structure of the second randomly tracted from biomass are described in the art. These
repeating monomer. processes include PHA extraction though the use of en-
[0035] The present invention further relates to an im- zymes, chemicals, mechanical means, and solvent ex-
proved process for extracting polyhydroxyalkanoate from traction, including extraction through the use of acetone
a biomass containing the polyhydroxyalkanoate compris- 35 and ketones. Without being limited by theory, it is be-
ing the steps of: lieved that the use of acetone under particular conditions
(a) maximizes both the yield and purity of the extracted
a) combining the biomass containing the polyhy- PHAs; and (b) minimizes the number of steps in the over-
droxyalkanoate with acetone to form a biomass liq- all extraction process and therefore at least partially ac-
uor wherein the biomass liquor is substantially free 40 complishes the objective of economical, commercial ex-
of water; traction of PHAs.
b) mixing the biomass liquor for from about 30 to [0042] The previously described embodiments of the
about 120 minutes at a temperature in the range of present invention may have many surprising advantages
from about 70°C to about 100 °C; over the current practice. For instance, the extraction
c) separating the polyhydroxyalkanoate from the bi- 45 process disclosed in the present invention may allow for
omass liquor to form a PHA-enriched liquor, wherein improved yield and/or purity and/or reduced costs. With-
the separating occurs by filtration at a temperature out being limited by theory, it is believed that the present
of at least about 70°C; invention may also allow for more efficient use of a con-
d) mixing the PHA-enriched liquor with water to form tinuous process due to an increase in process reliability
precipitated polyhydroxyalkanoate and an impure 50 and robustness associated with large scale production
solvent liquor, wherein the water is mixed with the according to the processes herein.
PHA-enriched liquor in the ratio of from about 5 parts [0043] Another surprising advantage of the present in-
to about 30 parts water to about one part polyhy- vention is the ability it confers to extract PHAs at lower
droxyalkanoate; and temperatures (below 150°C). Without being limited by
e) recovering the precipitated polyhydroxyalkanoate 55 theory, temperature considerations are important from a
from the impure solvent liquor. commercial standpoint, since the temperature at which
f) treating the precipitated polyhydroxyalkanoate a polymer is at least partially solubilized, and the time
with oxidizing agents, mild surfactants or mild required for adequate solubilization, can impact capital

4
 7 EP 1 781 721 B2 8

costs and product quality. For instance, PHAs that have ess of the present invention include, but are not limited
been subjected to lower temperatures for shorter periods to, apple, apricot, banana, cantaloupe, cherry, grape,
of time are typically of higher quality and increased use- kumquat, tangerine, tomato, and watermelon. Prefera-
fulness in downstream manufacturing processes. bly, the plants are genetically engineered to produce
[0044] The processes and methods herein may also 5 PHAs pursuant to the methods disclosed in Poirier, Y.,
include a wide variety of other variations. The processes D. E. Dennis, K. Klomparens and C. Somerville, "Poly-
and methods of the present invention are described in hydroxybutyrate, a biodegradable thermoplastic, pro-
detail hereinafter. duced in transgenic plants"’ SCIENCE, Vol. 256, pp.
[0045] The present invention relates to an improved 520-523 (1992); and/or U.S. Patent No. 5,650,555 to
process for extracting PHAs from a biomass using ace- 10 Michigan State University; issued July 22, 1997. Partic-
tone as a solvent, under selected process conditions. As ularly preferred plants are soybean, potato, corn, and
used herein, the phrase "extracting PHAs from a bio- coconut plants genetically engineered to produce PHAs;
mass", in addition to referring to the extraction of the more preferably soybean.
PHAs produced by a biomass which only produces a [0050] Bacteria useful in the present invention include
single type of PHA, also refers herein to the extraction 15 any genetically engineered bacteria designed to produce
of one or more types of PHA when the biomass produces PHAs, as well as bacteria which naturally produce PHAs.
more than one type of PHA. Examples of such bacteria include those disclosed in
[0046] The steps of this process are as follows: NOVEL BIODEGRADABLE MICROBIAL POLYMERS,
E.A. Dawes, ed., NATO ASI Series, Series E: Applied
I. Combining the biomass containing the polyhydroxyal- 20 Sciences-Vol. 186, Kluwer Academic Publishers (1990);
kanoate with a solvent to form a biomass liquor U.S. Pat. No. 5,292,860 to Kanegafuchi Kagaku Kogyo
Kabushiki Kaisha, issued Mar. 8, 1994; U.S. Pat. No.
a) Biomass Containing PHA 5,250,430 to Massachusetts Institute of Technology, is-
sued Oct. 5, 1993; U.S. Pat. No. 5,245,023 to Massa-
[0047] Polyhydroxyalkanoates ("PHAs") are extracted 25 chusetts Institute of Technology, issued Sep. 14, 1993;
via the process of the present invention from sources and/or U.S. Pat. No. 5,229,279 to Massachusetts Insti-
including, but not limited to, single-celled organisms, tute of Technology, issued Jul. 20, 1993.
such as bacteria or fungi, and higher organisms, such as [0051] It is preferable that the biomass contain a suf-
plants. These sources are collectively referred to herein ficient quantity of polyhydroxyalkanoate ("PHA") to make
as "biomass". While biomass may be comprised of wild- 30 the extraction process described in the present invention
type organisms, they are preferably genetically manipu- economically desirable. Preferably, the initial content of
lated species specifically designed for the production of PHAs in the biomass source material should be at least
particular PHAs of interest to the grower. Such geneti- about 20% of the total dry weight of the biomass; alter-
cally manipulated organisms are made through the natively at least 50%; alternatively, at least about 60%.
known processes of inserting foreign DNA, which is de- 35
rived from bacteria that naturally produce PHAs into an- b) Structurally flexible PHAs:
other organism.
[0048] The biomass containing the PHAs useful herein [0052] In one embodiment, the PHAs of the present
are preferably substantially dry. As used herein, "sub- invention are selected from those referred to herein as
stantially dry biomass" refers to biomass that contains 40 "structurally flexible" PHAs to underscore that the phys-
less than 5% water. Dry biomass is comprised of biomass ical disruption caused by the relatively high co-monomer
from which liquid has been removed using processes content and relatively long R-group chain length, make
including, but not limited to, spray or freeze drying, before them generally more ductile and harder to crystallize than
the extraction process is initiated. In one embodiment, PHAs that are characterized by lower co-monomer con-
the biomass contains less than 2% water, alternatively, 45 tent and shorter R-pendant groups (see U.S. Patents No.
the biomass contains less than 0.1% water, alternatively, 6,043,063 to Monsanto, issued March 28, 2000, and/or
the biomass contains no detectable level of water. 6,087,471 to Monsanto, issued July 11, 2000).
[0049] Plants useful as biomass in the present inven- [0053] The structurally flexible PHAs useful in the
tion include any genetically engineered plant designed present invention are in one embodiment comprised by
to produce PHAs. Preferred plants include agricultural 50 at least two randomly repeating monomer units, wherein
crops such as cereal grains, oilseeds and tuber plants; the first randomly repeating monomer unit has the struc-
more preferably avocado, barley, beet, broad bean, ture:
buckwheat, carrot, coconut, copra, corn (maize), cotton-
seed, gourd, lentil, lima bean, millet, mung bean, oat,
oilpalm, pea, peanut, potato, pumpkin, rapeseed (e.g., 55
canola), rice, sorghum, soybean, sugarbeet, sugar cane,
sunflower, sweet potato, tobacco, wheat, and yam. Such
genetically altered fruit-bearing plants useful in the proc-

5
 9 EP 1 781 721 B2 10

II. Mixing

[0057] The biomass liquor is then mixed for from about

10 to about 300, alternatively from about 10 to about 240
5 minutes at a temperature in the range of from about 70°C
to about 120°C. In one embodiment, the biomass liquor
is mixed at a temperature in the range of from about 70°C
and the second or higher randomly repeating monomer to about 100°C, alternatively in the range of from about
unit has the structure: 85°C to about 95°C. In one embodiment, the biomass
10 liquor is mixed for from about 30 to about 300 minutes;
alternatively from about 30 to about 120 minutes; alter-
natively from about 30 to about 60 minutes. Mixing can
be continuous or sporadic.
[0058] In one embodiment, the dry biomass is mixed
15 with acetone at a temperature in the range of from about
75°C to about 85°C, preferably about 80°C, for about 55
to about 65, preferably about 60, minutes.
wherein R is a C3 to a C7 alkyl or a mixture thereof; where- [0059] Mixing may be performed by any traditional
in from about 75 mol% to about 99 mol% of the randomly means of mixing compositions. For example, the mixing
repeating monomer units have the structure of the first 20 may be performed by using a mixing means selected
randomly repeating monomer unit, and from about 1 from propellers, turbines, screw conveyors, or mixtures
mol% to about 25 mol% of the randomly repeating mon- thereof. In one embodiment, the mixing is performed by
omer units have the structure of the second randomly using a plug flow concept with a screw conveyor.
repeating monomer unit. Such structurally flexible PHAs [0060] In one embodiment, the mixing is performed by
preferably have a melt temperature ("Tm") of about 80°C 25 using mixing means with a Power/Volume ratio of from
or higher. about 0.001 KW/m3 to about 100 KW/m3.
[0061] In a continuous process embodiment, a plug
c) Solvent: flow concept including a mechanical transportation sys-
tem such as a screw conveyor is used as the mixing
[0054] The biomass, containing the PHA, is combined 30 means.
with a solvent to form a biomass liquor. Although water
may be used as a solvent in certain applications, as used III. Separation
herein, the term "solvent" does not include water. The
solvent is acetone. [0062] The polyhydroxyalkanoate is then separated
[0055] In one embodiment, the biomass liquor contains 35 from the biomass liquor to form a PHA-enriched liquor.
the solvent and the polyhydroxyalkanoate in a ratio of at The separation occurs at a temperature of at least about
least about 5 parts solvent to about one part polyhydroxy- 50°C, preferably at least 70°C. Without being limited by
alkanoate. In another embodiment, the biomass liquor theory, it is believed that gelling occurs at temperatures
contains the solvent and the polyhydroxyalkanoate in a less than about 50°C resulting poor reliability/yield.
ratio of from about 5 to about 30 parts solvent to about 40 [0063] Means of separating the biomass from the bio-
one part polyhydroxyalkanoate. In another embodiment, mass liquor include filtration and/or centrifugation. In one
the biomass liquor contains the solvent and the polyhy- embodiment, hot filtration is used to separate the poly-
droxyalkanoate in a ratio of from about 10 parts solvent hydroxyalkanoate from the biomass liquor. As used here-
to about 1 part polyhydroxyalkanoate; alternatively in a in, the term "hot filtration" refers to filtering at a temper-
ratio of from about 20 parts solvent to about 1 part poly- 45 ature of at least about 40°C, preferably at least about
hydroxyalkanoate; alternatively from about 15 parts sol- 50°C. In one embodiment, hot centrifugation is used to
vent to about 1 part polyhydroxyalkanoate. In one em- separate the polyhydroxyalkanoate from the biomass liq-
bodiment, the biomass contains from about 3.2% to uor. As used herein, "hot centrifugation" refers to centrif-
about 9% of PHA in solution, alternatively the biomass ugation that occurs at a temperature of at least about
contains about 6.25% in solution. 50 40°C, preferably at least about 50°C. In one embodiment,
[0056] The biomass liquor contains less than about the filtration or centrifugation occurs at a temperature of
25% water; alternatively less than about 15%, alterna- at least about 70°C; alternatively at about 80°C, alterna-
tively less than 8%, alternatively less than about 5% wa- tively at about 90°C.
ter, alternatively less than about 2% water, alternatively, [0064] In one embodiment, high pressure centrifuges
the biomass liquor contains no measurable quantity of 55 are utilized in order to accommodate the higher temper-
water. atures and enhance the separation reliability.

6
 11 EP 1 781 721 B2 12

IV. Precipitation bine, high shear, and mixtures thereof. Where a propeller
is used, the P/V ratio may be from about 0.0001 to about
[0065] The PHA-enriched liquor is then mixed with wa- 100 and the resulting morphology of the PHA is typically
ter to form precipitated PHA and an impure solvent liquor. particles. Where a turbine is used, the P/V ratio may be
The water is combined with the PHA-enriched liquor in 5 from about 0.001 to about 1000 and the resulting mor-
the ratio of from at least 0.35 parts water to about one phoogy of the PHA is typically fibrous. Where High Shear
part solvent, alternatively, from about 0.35 to about 3 is used, the P/V ratio may be at least about 100, and may
parts water to about one part solvent, alternatively, from be a homogenizer. Where High Shear is used, the re-
about 1 parts water to about one part solvent. sulting morphology of the PHA is typically powder. With-
[0066] In one embodiment, the water is combined with 10 out being limited by theory, it is believed that mild mixing
the PHA-enriched liquor in the ratio of from about 0.75 with both radial and vertical flows should enable good
to about 1.5 parts water to about 1 part solvent. precipitation with few gelling issues.
[0067] In one embodiment, when the PHA-enriched [0074] The water addition rate during a batch process
liquor is mixed with the water, the PHA-enriched liquor may be greater than about 10 minutes of water addition.
is cooled to a temperature of from about 20°C to about 15 During a continuous process, the mixing of the water with
45°C. the PHA-enriched liquor should be such that it is in a
[0068] Without being limited by theory, it is believed similar ratio of water to enriched liquor. During a contin-
that too little water results in swelling/gelling and entrap- uous process, water may be added through a pump that
ment of solvents and/or other impurities in the gel. Further is capable of generating sufficient velocity to clear any
without being limited by theory, it is believed that excess 20 fine particles at the port of entry. Preferably, water injec-
water results in higher solvent recovery/recycling and tion through fine nozzles immersed in solution is avoided.
handling costs.
[0069] The PHA-enriched liquor and water may be V. Recovery
mixed together by a mixing means selected from propel-
lers, turbines, homogenizers, layers of water coated 25 [0075] The precipitated PHA is then recovered from
sheets, moving belts, high shear mixers, and combina- the impure solvent liquor. Filtration may be used to re-
tions thereof. Any tip speed and P/V (Power to Volume) covery the precipitated PHA from the impure solvent liq-
ratios are selected to obtain the desired product morphol- uor to produce recovered precipitated PHA and the re-
ogy. By using a propeller mixing means having both radial mainder impure solvent liquor.
and vertical mixing enables particles to be formed. By 30 [0076] In addition to filtration, the recovered precipitat-
using turbine mixing means, fibers or fibrous PHA prod- ed PHA may be squeezed and/or placed under pressure
uct may be formed. By using homogenizer mixing means, in order to remove any remainder impure solvent liquor.
fine particles that may agglomerate later may be formed. [0077] In addition to filtration and/or other recovery
[0070] Use of a propeller mixing means having a P/V means, the recovered precipitated PHA may then be
ratio of from about 0.0005 to about 1 result in particles 35 washed with a solvent selected from acetone, methyl
having a size of around 10 microns to around 2 mm. Use ethyl ketone, lower chain ketones, and mixtures thereof.
of a turbine type with a P/V ratio of from about 0.005 to
about 10 results in fibers having a size of less than about VI. Drying
5 mm.
[0071] The water may be added to the PHA-enriched 40 [0078] After recovery of the recovered precipitated
liquor or the PHA-enriched liquor may be added to the PHA, in one embodiment, the recovered precipitated
water. Without being limited by theory, it is believed that PHA is dried by traditional means to remove any remain-
this selection can impact the morphology of the resulting der impure solvent liquor.
PHA material.
[0072] When the PHA-enriched liquor is added to wa- 45 VI. Recycling of Solvent
ter, a thick chunk of agglomerate or fibrous spindle can
be obtained without mixing or with mild mixing. Use of a [0079] After the step of recovering the precipitated
turbine mixing means can result in fibers or fibrous spin- PHA from the impure solvent liquor, the solvent liquor
dles. The rate of addition of water and the tip-speed can may be recovered and recycled and/or re-used by tradi-
be varied to obtain small or large fibrous spindles. The 50 tional means in the processes and methods herein. The
use of a high shear homogenizer can help break up the wash solvent filtrate with less than 10% water can be
fibers or agglomerates into smaller particles. Another op- recycled directly to extraction and the filtrates with higher
tion is to add the PHA-enriched liquor to water coated than 10% (preferably higher than 5% water) can be dis-
surface or filters to enable precipitation of the PHA into tilled to recover acetone for recycle.
films, sheets, pulp, etc. 55
[0073] When the water is added to the PHA-enriched
liquor, the PHA-enriched liquor and the water can be
mixed together by a means selected from propeller, tur-

7
 13 EP 1 781 721 B2 14

VII. Optional Post-Treating with Oxidizing Agents or Sur- [0089] Therefore, the following examples further de-
factants scribe and demonstrate the preferred embodiments with-
in the scope of the present invention.
[0080] After the precipitated PHA is recovered, it may
be desirable to further post-treat the PHA with either an 5 EXAMPLES
oxidizing/bleaching agent or a surfactant in order to re-
move undesirable color bodies and/or odors. When used Example 1: Acetone-Water Process
herein, oxidizing agents may be used in the amount of
from a bout 0.0001 to about 0.5 parts oxidizing agent to [0090] To 100 Kg of dried biomass containing approx-
about 1 part PHA, alternatively about 0.01 part oxidizing 10 imately 60% PHA (Polyhydroxy Butyrate and Hexanoate
agent to about 1 part PHA. When peroxide is used, it is copolymer with about 9 mole % of Hexanoate), is added
typically 0.01 part active peroxide to 1 part PHA is used approximately 900 kg of acetone (recycle, wash or fresh
as a dilute form (i.e. dispersed in water). When used here- acetone with water content of 3%) with moderate mixing
in, surfactants may be used in the range of about 0.005 to form a slurry. The mixture of biomass and acetone
part surfactant to about 1 part PHA. 15 slurry is then heated to about 90°C and held for about 1
[0081] Oxidizing/bleaching agents useful herein in- hour to allow separation of PHA from the biomass to oc-
clude air, hydrogen peroxide (H2O2), hypochlorites, cur. This solution, held at the about 90°C temperature
bleach compounds including chlorine, bromine, and/or and pressure of approximately 3 Bar is then transferred
iodine oxidizing compounds, benzoyl peroxide, C9OBS, to the filter (Nutsche filter) or centrifuge under pressure.
perborates, and mixtures thereof. 20 The spent biomass (solid) is separated from the solution
[0082] Surfactants useful herein include amine oxide, containing PHA and acetone. The solution containing
AES, and other common surfactants, and mixtures there- PHA and acetone is then transferred to a precipitation
of. tank and simultaneously water is added at half the rate
[0083] Washing with surfactants and/or treating with of the PHA-Acetone solution. Approximately 450 kg of
oxidizing agents may result in removal of color bodies 25 water is used as a precipitating solvent. During precipi-
resulting in mild to significant color improvement (a whiter tation, moderate mixing with P/V of 2 KW/m3 is applied.
sample after treatment), removal of bio-odors, and/or re- The precipitated PHA with the solvent is transferred to
duction in impurities. another Nutsche filter or centrifuge and then the solvents
[0084] The oxidizing agents and/or surfactants can be are separated from the PHA to form a PHA cake and
used to treat the polyhydroxyalkanoate by washing the 30 used solvent/filtrate. The cake of PHA is pressed to min-
wet polymer with the oxidizing agent and/or surfactant e imize the solvents entrapped within PHA. The filtrate with
oxidizing agent (eg. H2O2) and/or by utilizing a dilute acetone and water (about 66% Acetone and 33% water)
solution during the drying of the polymer. may then be distilled to recover acetone for recycled use.
[0085] The use of oxidizing agents during the drying About 360 kg of fresh or recovered acetone is added to
of the Polyhydroxyalkanoate is especially useful if the 35 the wet PHA cake and used to wash the PHA cake with
impurities that are desired to be removed are known to gentle mixing in the filter. The filtrate is then separated
be oxidizable, (such as tri acetone amine impurities from from the wet PHA cake. The wet cake is then pressed to
the acetone-water extraction/precipitation process). remove as much as acetone and water as possible. The
filtrate after wash with acetone and containing about less
VIII. Optional Process Parameters 40 than 10% water can be used for extracting PHA. During
the washing step, optionally use about 0.01 part of active
[0086] As discussed above, depending on the type of H2O2 (diluted in water): 1 part PHA to enhance color and
morphology (flake, fiber, powder, film) desired in the pre- odor improvements. The wet PHA cake is dried through
cipitated PHA, process parameters can be altered to ob- a rotary drier under vacuum and 60°C. About 55 kg of
tain such morphologies. For instance, the (a) water ad- 45 dry PHA is produced.
dition order, rate, temperature and ratio along with (b)
type of mixing such as mild (propeller), moderate (tur- Example 2: Acetone-Water Process
bine) and high shear homogenizer define the morphology
of the precipitated polymer. [0091] About 13.05 grams of biomass containing about
[0087] Apart from that the method of precipitation (mix- 50 60% PHA (Polyhydroxy Butyrate and Hexanoate copol-
ing and water addition) can be used as tools to enable ymer 6.5 mole % Hexanoate) is mixed with 100 grams
the neat polymers morphology (flake, fiber, powder, film) of acetone. The mixture is added to a lab set containing
and enhance the purity of the product. a reaction chamber with a sintered metal filter capable
[0088] Although great care has been taken herein to of handling moderate temperature and pressure. The re-
provide guidance as to the selection of such parameters, 55 actor is then heated to 90°C for one hour. The reactor is
one of ordinary skill will recognize that the optimal range cooled to about 60°C and the PHA is filter extracted
of unit operating conditions or individual devices could through a 10 micron filter at the bottom of the reactor.
vary according to the type of raw biomass. The PHA is precipitated from the filtrate by adding to a

8
 15 EP 1 781 721 B2 16

heel of acetone-water mixture. Additional water is then with gentle mixing in the filter. The acetone is then sep-
added at a ratio of 0.5 parts of water for each 1.0 part of arated from the wet PHA cake by filtration. The wet cake
filtrate. The water and acetone are filtered from the pre- is then pressed to remove as much acetone as possible.
cipitated PHA using a Buchner funnel with #40 Whatman 2) About 600 kg of water is added to the wet PHA cake
filter paper. The filtered PHA is rinsed with acetone. The 5 and used to wash the PHA cake with gentle mixing in the
filtered PHA is spread out in a watch glass and allowed filter. The water is then separated from the wet PHA cake
to air dry overnight. A PHA yield of about 80 to 85% is by filtration. The wet cake is then pressed to remove as
observed. much water as possible. 3) About 600 kg of a 0.32%
solution of Alkyldimethyl Amine Oxide is added to the wet
Example 3: Acetone-Water Process with H2O2 treat- 10 PHA cake and used to wash the PHA cake with gentle
ment during drying process mixing in the filter. The Alkyldimethyl Amine Oxide is then
separated from the wet PHA cake by filtration. The wet
[0092] About 13.05 grams of biomass containing about cake is then pressed to remove as much Alkyldimethyl
60% PHA (Polyhydroxy Butyrate and Hexanoate copol- Amine Oxide as possible. 4) About 600 kg of water is
ymer 6.5 mole % Hexanoate) is mixed with 100 grams 15 added to the wet PHA cake and used to wash the PHA
of acetone. The mixture is added to a lab set containing cake with gentle mixing in the filter. The water is then
a reaction chamber with a sintered metal filter capable separated from the wet PHA cake by filtration. The wet
of handling moderate temperature and pressure. The re- cake is then pressed to remove as much water as pos-
actor is then heated to 90°C for one hour. The reactor is sible. The wet PHA cake is dried under vacuum and 60°C.
cooled to about 60°C and the PHA is filter extracted 20 About 122 kg of dry PHA is produced.
through a 10 micron filter at the bottom of the reactor.
The PHA is precipitated from the filtrate by adding to a
heel of acetone-water mixture. Additional water is then Claims
added at a ratio of 0.5 parts of water for each 1.0 part of
filtrate. The water and acetone are filtered from the pre- 25 1. An improved process for extracting polyhydroxyal-
cipitated PHA using a Buchner funnel with #40 Whatman kanoate from a biomass containing the polyhydroxy-
filter paper. The filtered PHA is rinsed with acetone and alkanoate comprising the steps of:
then with water. The wet cake is dried in a closed con-
tainer with some H2O2 solution added at the end of the a) combining the biomass containing the poly-
drying process (∼0.01 part H2O2 diluted in water: 1 PHA). 30 hydroxyalkanoate with acetone, as a solvent to
A PHA yield of about 80 to 85% is observed. form a biomass liquor wherein the biomass liq-
uor comprises less than 25% water;
Example 4: Acetone-Water Process with Amine Ox- b) mixing the biomass liquor for from 10 to 300
ide Wash during washing step minutes at a temperature in the range of from
35 70°C to 100°C;
[0093] To 222 Kg of dried biomass containing approx- c) separating the polyhydroxyalkanoate from the
imately 60% PHA (Polyhydroxy Butyrate and Hexanoate biomass liquor to form a PHA-enriched liquor,
copolymer with about 6.7 mole % of Hexanoate), is added wherein the separating occurs at a temperature
approximately 1800 kg of acetone with mixing to form a of at least 40°C;
slurry. The mixture of biomass and acetone slurry is then 40 d) mixing the PHA-enriched liquor with water to
heated to about 90°C and held for about 1 hour to allow form precipitated polyhydroxyalkanoate and an
separation of PHA from the biomass to occur. This solu- impure solvent liquor, wherein the water is mixed
tion, held at the about 90°C temperature and pressure with the PHA- enriched liquor in the ratio of from
of approximately 3 Bar is then transferred to the filter at least 3 parts water to one part polyhydroxy-
(Nutsche filter) under pressure. The spent biomass (sol- 45 alkanoate; and
id) is separated from the solution containing PHA and e) recovering the precipitated polyhydroxyal-
acetone. The solution containing PHA and acetone is kanoate from the impure solvent liquor.
then transferred to a precipitation tank containing a seat
of acetone and water while simultaneously water is add- 2. A process according to Claim 1 wherein the biomass
ed at half the rate of the PHA-Acetone solution. Approx- 50 liquor comprises the solvent and the polyhydroxyal-
imately 900 kg of water is used as a precipitating solvent. kanoate in a ratio of from at least 5 to 30 parts solvent
The precipitated PHA with the solvent is transferred to to one part polyhydroxyalkanoate.
another Nutsche filter and then the solvents are separat-
ed from the PHA to form a PHA cake and used solvent/ 3. A process according to any of the preceding claims,
filtrate. The cake of PHA is pressed to minimize the sol- 55 wherein during the step of mixing the biomass liquor
vents entrapped within PHA. Four washes are performed the mixing is performed by using mixing means se-
on the PHA cake. 1) About 600 kg of acetone is added lected from propellers, turbines, screw conveyor,
to the wet PHA cake and used to wash the PHA cake and mixtures thereof, preferably wherein the mixing

9
 17 EP 1 781 721 B2 18

means has a Power/Volume ratio of from 0.001 bilden, wobei die Biomassen-Flüssigkeit weni-
KW/m3 to 100 KW/m3. ger als 25% Wasser umfasst;
b) Mischen der Biomassen-Flüssigkeit über 10
4. A process according to any of the preceding claims, bis 300 Minuten bei einer Temperatur im Bereich
wherein the process is a continuous process wherein 5 von 70°C bis 100°C;
during the step of mixing the biomass liquor, the mix- c) Abtrennen des Polyhydroxyalkanoats aus der
ing is performed by using a plug flow concept with a Biomassen-Flüssigkeit zur Bildung einer PHA-
screw conveyor. angereicherten Flüssigkeit, wobei die Abtren-
nung bei einer Temperatur von mindestens
5. A process according to Claim 1 wherein the biomass 10 40°C stattfindet;
liquor comprises less than 8% water, preferably no d) Mischen der PHA-angereicherten Flüssigkeit
measurable quantity of water. mit Wasser, um ausgefälltes Polyhydroxyalka-
noat und eine unreine Lösungsmittelflüssigkeit
6. A process according to any of the preceding claims, zu bilden, wobei das Wasser mit der PHA-an-
wherein the separating the polyhydroxyalkanoate 15 gereicherten Flüssigkeit in einem Verhältnis von
from the biomass liquor is by filtration and/or centrif- mindestens 3 Teilen Wasser zu einem Teil Po-
ugation. lyhydroxyalkanoat gemischt wird; und
e) Gewinnen des ausgefällten Polyhydroxyalka-
7. A process according to any of the preceding claims noats aus der unreinen Lösungsmittelflüssig-
wherein the separating of the polyhydroxyalkanoate 20 keit.
from the biomass liquor occurs at a temperature of
at least 70°C, preferably 85°C. 2. Verfahren nach Anspruch 1, wobei die Biomassen-
Flüssigkeit das Lösungsmittel und das Polyhydro-
8. A process according to any of the preceding claims, xyalkanoat in einem Verhältnis von mindestens 5 bis
wherein in step (d) the water is added to the PHA- 25 30 Teilen Lösungsmittel zu einem Teil Polyhydro-
enriched liquor and preferably the PHA-enriched liq- xyalkanoat umfasst.
uor and water are mixed together by a means se-
lected from propeller, turbine, high shear, and mix- 3. Verfahren nach einem der vorangehenden Ansprü-
tures thereof. che, wobei während des Schrittes des Mischens der
30 Biomassen-Flüssigkeit das Mischen durchgeführt
9. A process according to any of Claims 1 through 7, wird unter Verwendung einer Mischeinrichtung, ge-
wherein in step (d) the PHA-enriched liquor is added wählt aus Propellern, Turbinen, Schneckenförderer
to the water and preferably the PHA-enriched liquor und Mischungen hiervon, wobei vorzugsweise die
and water are mixed together by a means selected Mischeinrichtung ein Leistungs/VolumenVerhältnis
from high shear, layers of water coated sheets or 35 von 0,001 KW/m3 bis 100 KW/m3 aufweist.
moving belts and mixtures thereof.
4. Verfahren nach einem der vorangehenden Ansprü-
10. A process according to Claim 1 wherein in the step che, wobei das Verfahren ein kontinuierliches Ver-
of mixing the PHA-enriched liquor with water to form fahren ist, wobei während des Schrittes des Mi-
precipitated polyhydroxyalkanoate and an impure 40 schens der Biomassen-Flüssigkeit das Mischen
solvent liquor, the PHA enriched liquor is cooled to durchgeführt wird unter Verwendung eines Kolben-
a temperature of from 20°C to 45°C and wherein in strömungs-Konzeptes mit einem Schneckenförde-
step (e), the precipitated polyhydroxyalkanoate re- rer.
covery from the impure solvent liquor is by filtration
and/or centrifugation to produce recovered precipi- 45 5. Verfahren nach Anspruch 1, wobei die Biomassen-
tated polyhydroxyalkanoate and remainder impure Flüssigkeit weniger als 8% Wasser, vorzugsweise
solvent liquor. keine messbare Menge an Wasser, umfasst.

6. Verfahren nach einem der vorangehenden Ansprü-

Patentansprüche 50 che, wobei das Abtrennen des Polyhydroxyalka-
noats aus der Biomassen-Flüssigkeit durch Filtration
1. Verbessertes Verfahren zur Extraktion von Polyhy- und/oder Zentrifugation stattfindet.
droxyalkanoat aus einer das Polyhydroxyalkanoat
enthaltenden Biomasse, umfassend die Schritte: 7. Verfahren nach einem der vorangehenden Ansprü-
55 che, wobei das Abtrennen des Polyhydroxyalka-
a) Kombinieren der das Polyhydroxyalkanoat noats aus der Biomassen-Flüssigkeit bei einer Tem-
enthaltenden Biomasse mit Aceton als ein Lö- peratur von mindestens 70°C, vorzugsweise 85°C,
sungsmittel, um eine Biomassen-Flüssigkeit zu stattfindet.

10
 19 EP 1 781 721 B2 20

8. Verfahren nach einem der vorangehenden Ansprü- 2. Procédé selon la revendication 1, dans lequel la li-
che, wobei beim Schritt (d) das Wasser der PHA- queur de biomasse comprend le solvant et le poly-
angereicherten Flüssigkeit zugesetzt wird und vor- hydroxyalcanoate à un rapport d’au moins 5 à 30
zugsweise die PHA-angereicherte Flüssigkeit und parties de solvant pour une partie de polyhydroxyal-
Wasser mittels einer Einrichtung, gewählt aus Pro- 5 canoate.
peller, Turbine, Hochscherung und Mischungen
hiervon, miteinander vermischt werden. 3. Procédé selon l’une quelconque des revendications
précédentes, dans lequel lors de l’étape de mélange
9. Verfahren nach einem der Ansprüche 1 bis 7, wobei de la liqueur de biomasse, le mélange est réalisé au
beim Schritt (d) die PHA-angereicherte Flüssigkeit 10 moyen d’un moyen de mélange choisi parmi des hé-
dem Wasser zugesetzt wird und vorzugsweise die lices, des turbines, un transporteur à vis et leur mé-
PHA-angereicherte Flüssigkeit und Wasser mittels lange, de préférence dans lequel le moyen de mé-
einer Einrichtung, gewählt aus Hochscherung, lange possède un rapport puissance/volume de
Schichten aus wasserbenetzten Lagen oder Bewe- 0,001 KW/m3 à 100 KW/m3
gungsbändern und Mischungen hiervon, miteinan- 15
der vernetzt werden. 4. Procédé selon l’une quelconque des revendications
précédentes, dans lequel le procédé est un procédé
10. Verfahren nach Anspruch 1, wobei beim Schritt des continu dans lequel lors de l’étape de mélange de la
Mischens der PHA-angereicherten Flüssigkeit mit liqueur de biomasse, le mélange est réalisé au
Wasser, um ausgefälltes Polyhydroxyalkanoat und 20 moyen d’un concept d’écoulement piston avec un
eine unreine Lösungsmittelflüssigkeit zu bilden, die transporteur à vis.
PHA-angereicherte Flüssigkeit auf eine Temperatur
von 20°C bis 45°C gekühlt wird, und wobei beim 5. Procédé selon la revendication 1, dans lequel la li-
Schritt (e) die Gewinnung von ausgefälltem Polyhy- queur de biomasse comprend moins de 8 % d’eau,
droxyalkanoat aus der unreinen Lösungsmittelflüs- 25 de préférence aucune quantité mesurable d’eau.
sigkeit durch Filtration und/oder Zentrifugation statt-
findet, um wiedergewonnenes ausgefälltes Polyhy- 6. Procédé selon l’une quelconque des revendications
droxyalkanoat und einen Rest an unreiner Lösungs- précédentes, dans lequel la séparation du polyhy-
mittelflüssigkeit zu erzeugen. droxyalcanoate de la liqueur de biomasse est réali-
30 sée par filtration et/ou centrifugation.

Revendications 7. Procédé selon l’une quelconque des revendications

précédentes, dans lequel la séparation du polyhy-
1. Procédé amélioré permettant d’extraire un polyhy- droxyalcanoate de la liqueur de biomasse est réali-
droxyalcanoate d’une biomasse contenant le poly- 35 sée à une température d’au moins 70 °C, de préfé-
hydroxyalcanoate, comprenant les étapes : rence de 85 °C.

a) la combinaison de la biomasse contenant le 8. Procédé selon l’une quelconque des revendications

polyhydroxyalcanoate avec l’acétone en tant précédentes, dans lequel dans l’étape (d) de l’eau
que solvant pour former une liqueur de biomas- 40 est ajoutée à la liqueur enrichie en PHA, et de pré-
se dans laquelle la liqueur de biomasse com- férence la liqueur enrichie en PHA et l’eau sont mé-
prend moins de 25 % d’eau ; langées par un moyen choisi parmi une hélice, une
b) le mélange de la liqueur de biomasse pendant turbine, un cisaillement élevé et leurs mélanges.
10 à 300 minutes à une température dans la
plage de 70 °C à 100 °C ; 45 9. Procédé selon l’une quelconque des revendications
c) la séparation du polyhydroxyalcanoate de la 1 à 7, dans lequel dans l’étape (d) la liqueur enrichie
liqueur de biomasse pour former une liqueur en- en PHA est ajoutée à l’eau et de préférence la liqueur
richie en PHA, dans laquelle la séparation est enrichie en PHA et l’eau sont mélangées par un
réalisée à une température d’au moins 40 °C ; moyen choisi parmi un cisaillement élevé, des cou-
d) le mélange de la liqueur enrichie en PHA avec 50 ches de feuilles recouvertes d’eau ou des courroies
de l’eau pour former un polyhydroxyalcanoate en déplacement et leurs mélanges.
précipité et une liqueur de solvant impur, dans
lequel l’eau est mélangée avec la liqueur enri- 10. Procédé selon la revendication 1, dans lequel dans
chie en PHA à un rapport d’au moins 3 parties l’étape de mélange de la liqueur enrichie en PHA
d’eau pour une partie de polyhydroxyalcanoate ; 55 avec l’eau pour former un polyhydroxyalcanoate pré-
et cipité et une liqueur de solvant impur, la liqueur en-
e) la récupération du polyhydroxyalcanoate pré- richie en PHA est refroidie jusqu’à une température
cipité de la liqueur de solvant impur. de 20 °C à 45 °C, et dans lequel dans l’étape (e), la

11
 21 EP 1 781 721 B2 22

récupération du polyhydroxyalcanoate précipité à

partir de la liqueur de solvant impur est réalisée par
filtration et/ou centrifugation pour produire un poly-
hydroxyalcanoate précipité récupéré et une liqueur
de solvant impur restante. 5

10

15

20

25

30

35

40

45

50

55

12
EP 1 781 721 B2

13
 EP 1 781 721 B2

REFERENCES CITED IN THE DESCRIPTION

This list of references cited by the applicant is for the reader’s convenience only. It does not form part of the European
patent document. Even though great care has been taken in compiling the references, errors or omissions cannot be
excluded and the EPO disclaims all liability in this regard.

Patent documents cited in the description

• US 5650555 A [0049] • US 5245023 A [0050]

• US 5292860 A, Kanegafuchi Kagaku Kogyo Ka- • US 5229279 A [0050]
bushiki Kaisha [0050] • US 6043063 A [0052]
• US 5250430 A [0050] • US 6087471 A [0052]

Non-patent literature cited in the description

• POIRIER, Y. ; D. E. DENNIS ; K. KLOMPARENS ; • NOVEL BIODEGRADABLE MICROBIAL POLY-

C. SOMERVILLE. Polyhydroxybutyrate, a biode- MERS. NATO ASI Series, Series E: Applied Scienc-
gradable thermoplastic, produced in transgenic es. Kluwer Academic Publishers, 1990, vol. 186
plants. SCIENCE, 1992, vol. 256, 520-523 [0049] [0050]

14