polyhydroxyalkanoates: biodegradable polymers & plastics ... · pdf...

Polyhydroxyalkanoates:

Biodegradable Polymers & Plastics from

Renewable Resources

Graz University of Technology, Austria Institute of Biotechnology and Biochemical Engineering

martin.koller@tugraz.at

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

Biodiesel

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!!!

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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 !

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