polyhydroxyalkanoates: biodegradable polymers & plastics ... · pdf...
TRANSCRIPT
Polyhydroxyalkanoates:
Biodegradable Polymers & Plastics from
Renewable Resources
Graz University of Technology, Austria Institute of Biotechnology and Biochemical Engineering
M. KOLLER, A. SALERNO, A. MUHR, R. ESSL, H. ANGERER, A. REITERER, E. RECHBERGER, K. MALLI, G. BRAUNEGG
October 17th to 19th, 2012, Portorož, Slovenia
1
The 20th Jubilee Conference on Materials and Technology
Content of the Presentation The „Plastic Situation“ Today
PHA Biopolyesters - a Sustainable Solution
Potential Applications of PHAs
Challenges in PHA Production: Downstream
Processing and Process Design
Available Feedstocks for PHA Production
Our Case Studies:
WHEYPOL Project: Surplus Whey as Feedstock
ANIMPOL Project: Animal Waste Lipids as Feedstock
Conclusions and Outlook 2
Nowadays, We Literally Live in the „Plastic Age“…
100 million tons
1,5 million tons
250 million tons
60 years ago
20 years ago
2010
330 million tons
2015
Figure: Rising amounts of plastics produced globally (Koller et al., 2011)
3
Quantities of Consumed Plastic Materials in Different Global Regions
80-120 kg / a
Developed Countries
(average pro capite)
2-15 kg / a
Emerging and Developing Countries
(average pro capite) Emerging and developing
countries
Developed and
industrialized countries
250 Mtons / a
World production & consumption of
Plastic Materials
1. Highly resistant polymeric materials
2. No natural degradation (landfil crisis!)
3. Insufficient performance of recycling systems
4. High risk connected to the thermal conversion of plastics by incineration (generation of toxines)
5. CO2 generation! Green house gases! Global warming!
TODAYS SITUATION: Polymers Predominately Deriving from Petro-Industry
5
It is Time to Switch.....
1. Fluctuation of petrol price is the major factor of uncertainty for global industry.
2. Advanced methods for tracing and discharging of crude oil exist, but finally fossil resouces are limited!
3. Since the year 2011: Instability of the political situation in many oil-exporting countries!! (Libya, Bahrain etc.). Future situation in Iran or Saudi Arabia??
6
Polyhydroxyalkanoates (PHAs) are biopolymers produced by a broad range of prokaryotes from renewable resources. They are the only family of „bioplastics“ entirely produced AND degraded by living cells!
PHAs: a Sustainable Solution!
The industrial implementation of PHAs has two major impacts:
•in replacing petrol based plastics (and reducing problems caused by them!)
•in solving industrial waste problems
7
PHAs can be selected as a sustainable solution for polymer industry:
1. Biobased, Biocompostible and Biodegradable („green plastics“)
2. Produced by living microorganisms
3. PHAs and their follow-up products can be processed to create a broad range of marketable products for a variety of applications
PHAs: a Sustainable Solution!
8
PHAs can be selected as a sustainable solution for polymer industry:
1. Biobased, Biocompostible and Biodegradable („green plastics“)
2. Produced by living microorganisms
3. PHAs and their follow-up products can be processed to create a broad range of marketable products for a variety of applications
PHAs: a Sustainable Solution!
9
Occurence and Composition of Polyhydroxyalkanoates (PHAs)
• Reserve Compounds for Carbon
and Energy; intracellularily produced by numerous procaryotic micro-organisms
• Production under condituions of Carbon surplus together with a Limitation of an essential Growth component (e.g.: N, PO4
3-, O2; Mg, S, K)
• Biologically produced and biologically degradable (kompostierbare) BIOPLASTICS
Electron microscopic picture of Cupriavidus necator DSM 545;
biopolymer content 60 to 70 wt.-%
Picture provided by Dr. E. Ingolić, ZFE-FELMI Graz
Koller et al., Macromolecular Bioscience 7, 218-226, 2007
11
Inclusions (PHA Granula)
12
The Cyclic Nature of Polyhydroxyalkanoate Metabolism Starting from Agricultural Waste Materials
Convertible carbon
Source: e.g.: Glucose
PHA –
Accumulation!!
Acetyl-CoA
Acetoacetyl-CoA
3-Hydroxybutyryl-CoA
Poly-(3-Hydroxybutyrate)
Pyruvate CO2
Catabolism via e.g.
2-Keto-3-Desoxy-6-
Phosphogluconate
Pathway (KDPG)
„Ideal“ growth conditions:
no limitations!
Growth of Biomass!!
3-Hydroxybutyrate
Acetoacetate
C3 - Unit
C2 - Unit
C4 - Unit
C4 - Unit
C4 - Unit
C4 - Unit
Change of nutrient conditions to growth (no limitations!)
Limitation
of growth
component
plus
surplus of
carbon
3-Ketothiolase
Acetoacetyl-CoA
Reductase
PHA
Synthase
Depolymerase
3HB-Dehydrogenase
Thiaphorase
Polysaccharides,
Oligosaccharides
Waste Lipids
Biodiesel Glycerol
ß-Oxydation
Raw materials
“White Biotechnology”
Accessible C-source
Upstream processing (hydrolysis)
Microorganisms as „cell factories“
(Archea, Bacteria, Fungi)
Bioproducts
fermentation process
Downsstream processing (separation
& purification)
Haloferax mediterranei Xanthomonas campestris
(PHA-biopolymer and
Polysacharide
production)
(Polysaccharide
production)
(PHA-biopolymer
production)
Bacillus megaterium
13
Chemical Structure of PHAs • Building blocks mainly chiral (important exception: 4-
Hydroxybutyrate); chiral building blocks are enantiomerically pure [always (R) – Konfiguration] !
• Most important representative: Poly-3-Hydroxybutyrate (PHB); Homopolyester of 3-Hydroxybutyrate → highly crystalline, brittle material, high melting point hempers most processing steps
• Remedy: Via production strategy change of the polyester composition by incorporation of different building blocks → Finetuning of product properties; Possibility to insert functional groups for post-synthetic chemical and/or enzymatic modification
Chiral
Center
Microbial Production of PHAs
PHAs can be also classified as homo- , co-, or terpolyester s depending
on:
Depending on the microbial production strain, PHAs can be divided in
2 large groups based on the number of carbon atoms in the monomer
units:
• C3-C5: short-chain-lengh PHA (scl)
• C6-C14: medium-chain-lengh PHA (mcl)
• Substrate and/or co-substrate
• Production strain
General chemical structure of
PHAs. The chiral center is
indicated by an asterisk (*)
Significance: Properties similar to Thermoplasts, Elastomeres or Latexes from
petrol-based industry. High dependence of properties on composition and
production strategy
PHAs: Property Dependance on Composition
16
scl-PHAs mcl-PHA
Khanna and Srivastava, 2005; Williams and Martin, 2002; Koller et al., 2010
PHAs can be selected as a sustainable solution for polymer industry:
1. Biobased, Biocompostible and Biodegradable (green plastic)
2. Produced by living microorganisms
3. PHAs and their follow-up products can be processed to create a broad range of marketable products for a variety of applications
PHAs: a Sustainable Solution!
17
Potential Applications of PHAs
• Biodiesel obtained by transesterification of PHAs with longer side chains (mcl-PHA) (sewage water)
18
•„Smart“ latexes and gels; thermosensitive adhesives; carriers for active compounds
Surgical Applications: Implants
Ongoing project „BioResorbable Implants for Children“ (BRIC) [Laura Bassi Center of Expertises; Austrian project]: Development of BioResorbable Implants for Children surgery (healing of femoral fractures). Coordinated by Medical University Graz, Austria; Prof. A.
Weinberg Now: Rat animal experiments; soon: lamb model Optimization of experimental in-vitro set-ups for degradation
tests for the implants
Literature: Artificial organs, artifical blood vessels, materials for wound treatment:
Application of PHAs
19
Chen et al., 2005 Sodian et al., 2000 Rokkanen et al., 2000
Surgical Applications: Implants
Ongoing project „BioResorbable Implants for Children“ (BRIC) [Laura Bassi Center of Expertises; Austrian project]: Development of BioResorbable Implants for Children surgery (healing of femoral fractures). Coordinated by Medical University Graz, Austria; Prof. A.
Weinberg Now: Rat animal experiments; soon: lamb model Optimization of experimental in-vitro set-ups for degradation
tests for the implants
Literature: Artificial organs, artifical blood vessels, materials for wound treatment:
Application of PHAs
20
The Entire Process..
Renewable Raw Material. Here: Sugar cane
The Entire Process..
Selection of powerful microbial production strain
The Entire Process..
Evaluation of optimimum cultivation conditions
The Entire Process..
Bacterial cell harbouring PHA granules
The Entire Process..
Biosynthesis under controlled and optimized conditions (bioreactor)
The Entire Process..
PHA granulate
Bacterial cell harbouring PHA granules
Downstream processing for PHA- biopolymer recovery
The Entire Process..
PHA granulate
The Entire Process..
Biodegradation (Composting)
The Entire Process..
Final products of the oxidative catabolism:
carbon dioxyde and water!
1. The cost of downstream processing for recovery of PHA from biomass
2. The process design (discontinuous vs. continuous fermentation mode)
3. The selection of raw materials
Obstacles for the Market Penetration of PHAs
The production costs of PHA must be in the same range as the „classical“ petrochemical competitors on the plastic market (PP, PE, PET etc.)
Hence, they have to be minimized despite the instable market price for crude mineral oil
This can be accomplished by optimizing:
30
Obstacles for the Market Penetration of PHAs
The production costs of PHA must be in the same range as the „classical“ petrochemical competitors on the plastic market (PP, PE, PET etc.)
Hence, they have to be minimized despite the instable market price for crude mineral oil
This can be accomplished by optimizing:
31
The cost of downstream processing for recovery of PHA from biomass
@ Downstream Processing
• Extraction of PHA from surrounding cell mass
• Often high throughput of (even toxic!) solvents and high energy requirements!
• Screening of alternative solvents and efficient extraction strategies needed
• Mechanical cell disruption, chemical or enzymatic digestion of non-PHA cell mass
32
Released PHA
Granules
(often still
surrounded by
membrane) Crude extracted PHA
(high purity)
+ Lipids
+ +
Chemical or enzymatic digestion of
non-PHA cell mass
or
Mechanical disruption of cells
or
Disruption of cells with high
intracellular osmotic pressure in
hypotonic media
PHA-rich Biomass
Removal of Lipids with
Organic Solvents or sCO2
(Degreasing)
PHA-free Cells
Degreased PHA-rich
Biomass
Extraction with organic
solvent (e.g. Chloroform)
Biomass Digestate
(Cell Debris)
Cell Harvest, Separation
from Aqueous
Supernatant, Drying
34
Downstream Processing of Biomass for PHA Isolation:
Extraction
Industrial scale: e.g.
Podbielniak Extractor
Extraction on laboratory scale: e.g.
Soxleth Extractor
35
Future of PHA Isolation from Biomass
Alternative: Investigation of „Green Solvents“ that are easy to recycle (e.g. lactic acid esters)
Avoiding of chlorinated solvents like chloroform!!
PHBISA Process Brazil: PHA extraction via the fusel alcohol fraction (iso-pentanol) from the destillative Bioethanol production
Taking profit of the high intracellular pressure of osmophilic production strains (example: Haloferax mediterranei) → simple release of intracellular PHA-granula in hypotonic medium (deionized water); low purity; membranes of granules remain intact!
Novel Strategy for PHA Extraction Developed at TU Graz
36
Extraction at temperature above
the solvent´s boiling point in a first
vessel. System under Pressure
Solvent: Classial „non-PHA solvent“
at room temperature!
Pressure release by opening of
valve; residual biomass remains in
filter, PHA dissolved in solvent
passes through to vessel 2
Precipitation of polymer in vessel 2
(cooling)
Obstacles for the Market Penetration of PHAs
The production costs of PHA must be in the same range as the „classical“ petrochemical competitors on the plastic market (PP, PE, PET etc.)
Hence, they have to be minimized despite the instable market price for crude mineral oil
This can be accomplished by optimizing:
37
The process design (discontinuous vs. continuous fermentation mode)
@ Process Design: PHA Production at Graz University of Technology
Batch vs. Continuous Production Mode
38
5-Stage Continuous Process
• Microbial growth in first vessel („ideal“ cultivation conditions; autocatalytic process)
• Vessel 2 to 5: PHA-production (Carbon supply; limitation of essential growth component)
• Vessel 2 to 5 are process engineering substitute for a tubular reactor! According to PHA production kinetics (linear increase of PHA concentration)!
• High productivities!
• Constant product quality (Molecular mass, Thermoanalytical data) during steady state
39
40
41
Atlić, A. et al. (2011). Continuous Production of Poly([(R]-3-hydroxybutyrate) by Cupriavidus
necator in a Multistage Bioreactor Cascade, Applied Microbiology and Biotechnology 91: 295-304
Obstacles for the Market Penetration of PHAs
The production costs of PHA must be in the same range as the „classical“ petrochemical competitors on the plastic market (PP, PE, PET etc.)
Hence, they have to be minimized despite the instable market price for crude mineral oil
This can be accomplished by optimizing:
42
The selection of raw materials
Carbon-Rich Waste Streams Selection
No interference with food- or feed applications!!!
43
1. Whey from dairy industry (Lactose): EU-FP5 PROJECT WHEYPOL (Dec. 2001 to Dec. 2004; coordinated by Graz University of Technology)
2. Crude glycerol phase from the biodiesel production (Glycerol) EU-FP5 PROJECT BIODIEPRO (Jan. 2003 to Dec. 2005; coordinated by ARGENT Energy; Graz University of Technology as partner)
3. Molasses from the sugar industry (Sucrose) (Bilateral project with Brazilian company PHBISA/Copersugar)
4. Animal Derived Waste Lipids (EU-FP7 PROJECT ANIMPOL): ongoing since 2010; coordinated by Graz University of Technology)
Our Selected Alternative Carbon Sources:
44
Our Case Study 1: FP5 Project WHEYPOL
The WHEYPOL project developed a
sustainable and sound process for the conversion of surplus whey from dairy industry to PHA biopolyesters
in order to create a viable strategy that enables the production of PHAs in Europe in future
45
Significance of the WHEYPOL Project
Application of whey lactose (D-gluco-pyranose-4-ß-D-galactopyranoside) from dairy industry:
Animal feed
Sweets
Food processing
Baby food
Laxatives
Pharmaceutical matrices
But: annually 13,462.000 t of surplus whey in Europe (620.000 t lactose)!
Global amounts: up to 1.60*108 t (data for 2008); annual increase about 2%!!
Ecological problem; polluting whey (high COD and BOD!) partly disposed in rivers or sea
2001: EU – project WHEYPOL (G5RD-CT-2001-00591): application of surplus whey from Italian dairy industry as substrate for PHA biopolyester production; Coordinated by Graz University of Technology
46
Composition of Different Types of Whey (Braunegg et al., 2007)
Compound
[% (w/w)]
Sweet
Whey
Fermented
Whey
Whey Permeate
(Substrate for
Biotechnology!)
Whey
Retentate
(Marketable
Proteins)
Lactose 4.7–4.9 4.5–4.9 23 14
Lactic acid traces 0.5 - -
Proteins 0.75–1.1 0.45 0.75 13
Lipids 0.15–0.2 traces - 3-4
Inorganic compounds
(minerals like e.g. calcium)
ca. 7 6-7 ca. 27 ca. 7
47
WHEYPOL: PHA Production from Surplus Whey
http://news.cec.eu.int/comm/research/industrial_technologies/ articles/article_805_en.html
Dairy industry waste is a potential source of biologically-produced polymers with commercial applications in packaging. WHEYPOL developed a cost-effective method to tap this abundant and sustainable resource.
Whey production in Europe: 40,420.800 tons/y Surplus WHEY: 13,462.000 tons/y
Lactose: 619.250 tons /a 205.000 t PHA/a 48
The Consortium of WHEYPOL The research was performed by a consortium from 6 European countries: close cooperation of 6 academic and 3 industrial partners from 5 countries! Academic Partners:
Partner Partner Logo Key Researcher Main Roles Country
Graz University of Technology
Prof. Gerhart Braunegg, Prof. Michael Narodoslawsky, Prof. Rolf Marr
Coordination; Biotechnological production of PHA biopolyesters (Institute of Biotechnology and Biochemical Engineering); Downstream Processing; Life Cycle Assessment, Cleaner production studies; (Institute of Process and Particle Engineering)
Austria
Università di Padova
Prof. Sergio Casella Microbiology, Genetics Italy
Slovak Academy of Science
Prof. Ivan Chodak Characterization of PHAs and follw-up products Slovakia
Università di Pisa Prof. Emo Chiellini Characterization of PHAs; formulation of PHA-based composites and blends
Italy
Polish Academy of Science
Prof. Marek Kowalczuk Characterization of PHA and derived composites and blends
Poland
National Institute of Chemistry
Dr. Andrej Kržan Characterization of PHA and derived composites and blends
Slovenia
49
Industrial Waste-Streams from…
Biotechnological conversion of waste streams from dairy industry (surplus whey) towards PHA biopolyesters.
Latterie Vicentine Soc.Coop. A R.L. Large Italian dairy company. Key representative: Mr. Luigi Sibilin
50
Additional Industrial Partners:
BDI - BioEnergy International AG Large Austrian company specialized in construction of technical plants (biodiesel). Role in WHEYPOL: Process design & Engineering Key representative: Dr. Edgar Ahn
Idroplax srl, Italy. Representative of Polymer Industry! Interested in switching to bioplastics. Role in WHEYPOL: Processing of biopolymers on large scale Key representatives: Mr. Luca Landini; Mr. Bayan Giltsoff
How industry can support and optimize academic research!
51
Biotechnological Example: Fermentation Pattern for PHA Production from Hydrolyzed Whey
Lactose on a Highly Saline Nutrient Medium by the Archaeon Haloferax mediterranei
0
2
4
6
8
10
12
14
0 50 100Time [h]
[g/L
]
Glucose Galactose
3-PHA Protein
Limitation of
growth
component
(nitrogen,
phosphate):
residual
biomass
(expressed as
protein)
concentration
remains
constant,
carbon flux
towards PHA
accumulation)
52 Koller et al., Biomacromolecules 6, 561-565, 2005
Process Parameters Values
Cell Dry Mass 11.0 [g/L]
PHA 5.5 [g/L]
Residual Biomass 5.5 [g/L]
PHA / CDM 49.6 [%]
µ max. 0.11 [1/h]
Volumetric Productivity 0.05 [g/Lh]
Yield PHA / Whey sugars 0.33 [g/g]
Main Results:
53
Koller et al., Biomacromolecules 6, 561-565, 2005
The WHEYPOL Process: Economic Assessment
Koller et al., Macromolecular Bioscience 7, 218-226, 2007
Choi and Lee
1997
Choi and Lee
1999
Reddy et al.,
1999 Haloferax
mediterranei,
Koller et al.
2007
Hydrogenophaga
pseudoflava,
Koller et al. 2007
Pseudomonas
hydrogenovora,
Koller et al. 2007
Beneficial Combined Effects
Waste stream (Whey) as
Raw Material
High-value Copolyester
from „simple“ carbon-source
Lactose (no addition of
precursor)
Insterile „septic“ Process
possible; safes energy for
sterilization (extreme
halophilic!)
Product isolation: simple
release of PHA granula in
deionized water (high
intracellular osmotic pressure)
54
Project Start: January 1st, 2010
Entire Project Volume: € 3,7 Mio.; EU contribution: € 2,9 Mio
Coordinated by Graz University of Technology, Austria
Example 2: FP7 Project ANIMPOL
„Biotechnological conversion of carbon containing wastes for eco-efficient production of high added value products”
55
FP7 Project ANIMPOL
The Animpol project aims at the
sustainable and value added conversion of waste-lipids from animal processing industry (waste streams from
slaughterhouses, the animal rendering industry and
waste fractions from conventional biodiesel production)
in order to create a viable strategy that enables the production of PHAs in Europe in future.
Bring together waste producers from animal processing industry and biofuel industry with the polymer industry.
Development of an integrated, sound industrial process!
56
MICROBIAL PHA PRODUCTION (group 1 and group 2 production strains)
Downstream Processing RECOVERY OF PHA FROM
BIOMASS
Waste Fraction
Hydrolysis RESIDUAL BIOMASS Purification/Refining
PHA
WASTE LIPIDS Transesterification
MIX BIODIESEL-GLYCEROL Separation
BIOFUEL (RME) GLYCEROL LIQUID PHASE (GLP)
Proteins Lipids
57
Amounts of Waste in EU Relevant for ANIMPOL
ANIMAL WASTE LIPIDS 500.000 t/y
CRUDE GLYCEROL 265.000
metric tons/year
BIODIESEL
CATALLYTICALLY ACTIVE
BIOMASS (0.4-0.5 g/g)
PHA 120.000 t (0.3 g/g)
SATURATED FRACTION
50.000 t/year
UNSATURATED FRACTION
PHA 35.000 t (0.7 g/g)
Excellent 2nd generation Biofuel!
58
The Consortium of ANIMPOL The research is performed by a consortium from 6 European countries: close cooperation of 7 academic and 4 industrial partners from 7 countries
Academic Partners:
Partner Partner Logo
Key Researcher Main Roles Country
Graz University of Technology
Dr. Martin Koller, Prof. Michael Narodoslawsky, Prof. Hans Schnitzer
Coordination; Biotechnological production of PHA biopolyesters (Institute of Biotechnology and Biochemical Engineering); Life Cycle Assessment, Cleaner production studies; Process engineering (Institute of Process and Particle Engineering)
Austria
Università di Padova Prof. Sergio Casella Microbiology, Genetics Italy
University of Zagreb Prof. Predrag Horvat Mathematical modeling of bioprocesses Croatia
University of Graz Prof. Martin Mittelbach Enhanced transesterification of waset animal lipids; assessment of composition and quality of raw materials
Austria
Università di Pisa Prof. Emo Chiellini Characterization of PHAs; formulation of PHA-based composites and blends
Italy
Polish Academy of Science
Prof. Marek Kowalczuk Characterization of PHA and derived composites and blends
Poland
National Institute of Chemistry
Dr. Andrej Kržan Characterization of PHA and derived composites and blends
Slovenia 60
Industrial Waste-Streams from… Biotechnological conversion of waste streams from two industrial branches towards PHA biopolyesters.
U. Reistenhofer GesmbH, Austria. Slaughtering industry: lipid rich animal residues. Key representative: Mr. Thomas Reistenhofer
Argent Energy, Great Britain. Large biodiesel producer from tallow (highly saturated biodiesel fractions) and waste cooking oil; delivers saturated biodiesel fraction and crude glycerol phase. Key representative: Dr. Mike Scott
61
Additional Industrial Partners:
Argus Umweltbiotechnologie GmbH, Germany. Scale-up of industrial process from lab scale (from 1L to industrial scale 70000 L). Role in ANIMPOL: development of sustainable Downstream Processing Key representative: Dr. Horst Niebelschütz
TERMOPLAST srl, Italy. Representative of Polymer Industry! Interested in switching to bioplastics. Key representative: Dr. Maurizio Malossi
How industry can support and optimize academic research!
62
• Advisory Board members are no beneficiaries of the project; they give advice in how to proceed with the activities
Advisory Board of Companies Acting as an „Enduser Group“
1. Novamont, Italy: biodegradables
2. ChemTex Italia (gruppo Mossi & Ghissolfi; Italy): biobased products
3. KRKA, Slovenia: large scale fermentations
63
4. Eksportera UAB, Lithuania: long expertise in by-product business
The Holistic Nature of Animpol
64
Biotechnological Example: Fermentation Pattern for scl-PHA Production from Animal-Derived, Saturated Biodiesel. Production strain Cupriavidus nector. Additional: 3HV Precursor Valeric Acid
65
0,00
5,00
10,00
15,00
20,00
25,00
30,00
35,00
40,00
45,00
50,00
0 10 20 30 40 50 60 70 80 90 100
co
nc
en
tra
tio
n [
g/L
]
time [h]
CDM
PHA
Nitrogen limitation at t = 65 h
66
0
10
20
30
40
50
60
70
16 19 24 40 44 46 49 64 66 69 72 75 88 90 91
[wt.
-%]
time [h]
PHA/CDM
3HV/PHA
Nitrogen limitation at t = 65 h
Addition of 3HV precursor
valeric acid from t = 69 h
Biotechnological Example: Fermentation Pattern for scl-PHA Production from Animal-Derived, Saturated Biodiesel. Production strain Cupriavidus necator. Additional: 3HV Precursor Valeric Acid
Process Parameters Values
Cell Dry Mass 47.2 [g/L]
PHA 30.0 [g/L]
Residual Biomass (non-PHA Biomass) 21.1 [g/L]
PHA / CDM 63.6 [%]
µ max. 0.04 [1/h]
3HV / PHA 4.3 [mol-%]
Volumetric Productivity (Accumulation Phase) 1.08 [g/Lh]
Specific Volumetric Productivity (Accumulation Phase) 0.37 [g/gh]
Yield (Biomass / Biodiesel) 0.6 – 0.7 [g/g]
Main Results:
67
68
The Crude Product:
Poly-(3HB-co-4.3%-3HV)
Biodegradable Latexes from Animal-Derived Waste: Biosynthesis and Characterization of mcl-PHA
Accumulated by Ps. citronellolis
69
Submitted to be published in Reactive and Functional Polymers
mcl-PHA from Saturated Biodiesel by Ps. citronellolis : Polymer Composition
70
• According to GC-FID analysis, the obtained biopolyester predominantly consists of
• 3-hydroxyoctanoate (C8) and 3-hydroxdecanoate (C10), and,
• to a minor extent, 3-hydroxydodecanoate (C12), 3-hydroxynonanoate (C9), 3-hydroxyhexanoate (C6), and 3-hydroxyheptanoate (C7) monomers.
• This was confirmed by 1H-NMR, also evidencing the occurrence of low quantities of unsaturated and 3-hydroxyvalerate (C5) building blocks.
mcl-PHA from Animal-Derived Biodiesel by Ps. citronellolis: Concentrations of Building Blocks during the Cultivation
(GC-FID)
71
Nitrogen limitation
mcl-PHA from Animal-Derived Biodiesel by Ps. citronellolis: Shares of Building Blocks in Polymer during the Cultivation
(GC-FID)
72
Nitrogen limitation
73
Process Parameters Values
PHA 3.0 [g/L]
Catalytically Active Biomass (Protein) 11.2 [g/L]
Sum Protein + PHA 14.2 [g/L]
PHA / CDM 20.1 [%]
µ max. 0.08 [1/h]
Volumetric Productivity (Accumulation phase) 0.07 [g/Lh]
Specific Volumetric Productivity (Accumulation Phase) 0.02 [g/gL]
Yield (Biomass / Biodiesel) 0.5 – 0.6 [g/g]
Main Results:
Comparison of Material Properties scl-PHA mcl-PHA
Production strain Cupriavidus necator Ps. citronellolis
Composition P(3HB-co-4.3%-3HV) Mainly 3HO and 3HD; to minor extend: 3HDD, 3HN, 3Hx, 3Hp
Tm [°C] 163.0 48.6
δHm [J/g] 78.9 7.1
Xc [%] 54.0 12.3
Tg [°C] -2.8 -46.9
Td [°C] 282 296
Mw [kDa] 318 66
Mn [kDa] 233 35
Pi (Polydispersity) 1.4 1.9
Mw via SEC-MALS [kDa] 18.7 n.d.
Mn via SEC-MALS [kDa] 27.0 n.d.
74
SEC-MALS for scl-PHA: group Dr. Andrej Kržan, National Institute of Chemistry, Ljubljana, SLO
Measurements scl-PHA: group Prof. Emo Chiellini, University of Pisa, I
Measurements mcl-PHA: group Prof. Marek Kowalczuk, Polish Academy of Science, PL
1. General Impact: • solutions for waste problems arising on local
scales that can be applied for all Europe.
2. Transitional Impact: • creation of ecological and economic benefits by
converting waste into value-added materials
3. Socioeconomic Impact: • new jobs directly in the industrial branches
and high-qualified scientific jobs in academia.
Potential Impact of the Projects
PHA-Biopolymer Production can Become Economically Competitive by:
• Utilization of locally available waste materials
• Integration of PHA production into existing production line
• Alternative extraction methods
Especially for WHEYPOL (and soon for ANIMPOL):
Data for designing a pilot plant to be integrated in
large dairies are available!
Willingness of responsible policy-makers from
waste-generating industrial branches and from
polymer industry to break new ground in
sustainable production is needed!!!
77
Is there a Need for „White Biotechnology“ for Production of
Biopolymers, Biofuels and Biochemicals??
January 2007 to June 2008: Price jumped by more than 100% and surmounted 130 US-$ per barrel
January 2009: Back to less than 40 US-$ per barrel
June 2010: Again 77 US-$ per barrel!!
April 2011: 127 US-$ per barrel!!
October 15th, 2012: 116 US-$ per barrel!!
TOMORROW: WHO KNOWS????? Increasing uncertainties in global political situation! 78
Linear increase
Exponential increase
Acknowledgements
The audience and the organizers of The 20th Jubilee
Conference on Materials and Technology !
79