Sustainable and Efficient Production of Biopolymers from Industrial Waste

Streams

Sustainable and Efficient Production of Biopolymers from Industrial Waste

Streams

Graz University of Technology, AustriaInstitute of Biotechnology and Biochemical Engineering martin.koller@tugraz.at

Martin Koller, Anna Salerno, Alexander Muhr, Robert Essl, Angelika Reiterer, Gerhart Braunegg

May 2nd to 4th, 2012, Bregenz, Austria

15th European Roundtable on Sustainable Consumption and Production (15th ERSCP)

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Content of the Presentation The „Plastic Situation“ today

„White Biotechnology“

„Green Plastics“??

PHA Biopolyesters - a sustainable solution

Potential Applications of PHAs

Available Feedstocks for PHA Production

Our case studies:

WHEYPOL Project: Surplus whey as feedstock

ANIMPOL Project: Animal waste lipids as feedstock

Conclusions

Outlook: Is there a Need for „White Biotechnology“ for Production of Biopolymers? 2

Quantities of Utilized Plastic Materials in Different Global Regions

80-120 kg / aDeveloped &

Industrialized Countries(average consumption per person)

80-120 kg / aDeveloped &

Industrialized Countries(average consumption per person)

250 Mtons / aWorld Production & Consumption

of Plastic Materials

250 Mtons / aWorld Production & Consumption

of Plastic Materials

2-15 kg / a

Emerging and Developing Countries

(average consumption per person)

2-15 kg / a

Emerging and Developing Countries

(average consumption per person)

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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

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It is Time to Switch.....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 the fossil resouces are limited.

3. Year 2011: Instability of the political situation in many oil-exporting countries!! (Libya, Bahrain etc.). Future situation in Iran or Saudi Arabia??

4. The degradation products of these materials cause green house effect and global warming.

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Global Carbon Cycle (based on Narayan, 2002; Koller et al., 2011)

Result: a Balancing Problem!But: The Balancing Problem can be solved!6

Raw materials

““White Biotechnology”White Biotechnology”

Accessible C-source

Upstream processing (hydrolysis)

Microorganisms as „cell factories“

(Archea, Bacteria, Fungi)

Bioproducts

(fermentative process)

Downsstream processing

(separation &

purification)

Haloferax mediterraneiXanthomonas campestris

(PHA-biopolymer and Polysacharide production)

(Polysaccharide production)

(PHA-biopolymer production)

Bacillus megaterium

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What Characterizes a „GREEN“ Plastic?What Characterizes a „GREEN“ Plastic?

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„„Biodegradable“Biodegradable“The 90% of the carbon of the plastic is metabolized within 180 days. (standardized norm EN-13432)

„„Compostable“Compostable“

If not more than 10% of the plastic material remain in a sieve of 2mm pore size after 180 days of composting .(standardized norm EN-13432)

Using standardized methods for assessing the ecotoxicity of the (plastic) material, it must not feature any negative impact on living organisms or the involved environment.(standardized norm ISO 10993) „„Biocompatible“Biocompatible“

The production of the building blocks is based on renewable resources; the polymerization of the monomers may occur chemically or biotechnologically.

„„Biobased“Biobased“

Criteria for Talking about „GREEN“ PlasticsCriteria for Talking about „GREEN“ Plastics

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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!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.10

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!PHAs: a Sustainable Solution!

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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!PHAs: a Sustainable Solution!

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PHAs: Microbial Reserve Compounds for Carbon and Energy PHAs: Microbial Reserve Compounds for Carbon and Energy

PHAs serve as a storage materials for carbon and energy for the microorganism:

• produced: under conditions of high intracellular energy charge: carbon surplus together with a limitation of an essential growth component (nitrogen, phosphate).

• metabolised: under condition of starvation (low intracellular energy charge), PHAs are re-utilized by the microbial cell and converted to H2O and CO2 as the final products of their oxydative break down.

Electrone microscope picture of Cupriavidus necator DSM 545; PHA content in cells 60 to 70 wt.-%; Picture by Dr. E. Ingolić, ZFE-FELMI Graz

Koller et al., Macromolecular Bioscience 7, 2007

PHAs provide an advantage for microbial surviving!

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PHA Production at Graz University of TechnologyPHA Production at Graz University of Technology

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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!PHAs: a Sustainable Solution!

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Potential Applications of PHAs

• Biodiesel obtained by transesterification of PHAs with longer side chains (mcl-PHA) (sewage water)

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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

Artificial organs, artifical blood vessels, materials for wound treatment:

Application of PHAs Application of PHAs

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1.The selection of raw materials

2.The cost of downstream processing for isolation of PHA from biomass

3.The process design (continuous fermentation mode)

Obstacles in the MarketObstacles in the Market Penetration of Penetration of PHAsPHAs

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:

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Carbon-Rich Waste Streams Selection

No interference with food- or feed applications!!!

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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:

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Our Case Study 1: FP5 Project WHEYPOL

The WHEYPOL project developed asustainable and sound process for the conversion of surplus whey from dairy industry to PHA biopolyestersin order to create a viable strategy that enables the production of PHAs in Europe in future.

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Significance of the WHEYPOL ProjectApplication 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; Coordination at Graz University of Technology 22

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/ySurplus WHEY: 13,462.000 tons/y

Lactose: 619.250 tons /a 205.000 t PHA/a23

The Structure of WHEYPOLThe 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

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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. Sibilin

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Additional Industrial Partners:

BDI - BioEnergy International AGLarge Austrian company specialized in construction of technical plants (biodiesel). Role in WHEYPOL: Process design & EngineeringKey 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

How industry can support and optimize academic research!

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Biotechnological Example: Fermentation Pattern for PHA Production from Hydrolyzed Whey Lactose

0

2

4

6

8

10

12

14

0 50 100Time [h]

[g/L

]

Glucose Galactose3-PHA Protein

Limitation of growth component (nitrogen, phosphate): residual biomass (expressed as protein) concentration remains constant, carbon flux towards PHA accumulation)

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Process Parameters ValuesCell 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:

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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)

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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”

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FP7 Project ANIMPOLThe 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!

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MICROBIAL PHA PRODUCTION(group 1 and group 2 production strains)

Downstream ProcessingRECOVERY OF PHA FROM

BIOMASS

Waste Fraction

HydrolysisRESIDUAL BIOMASS

Purification/RefiningPHA

WASTE LIPIDSTransesterification

MIX BIODIESEL-GLYCEROLSeparation

BIOFUEL (RME)

GLYCEROL LIQUID PHASE (GLP)

Proteins Lipids

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Amounts of Waste in EU Significant for ANIMPOL

ANIMAL WASTE LIPIDS

500.000 t/y

CRUDE GLYCEROL 265.000

metric tons/year

BIODIESEL

CATALLYTICALLY

ACTIVE BIOMASS

(0.4-0.5g/g)

PHA120.000 t(0.3g/g)

SATURATEDFRACTION

50.000 t/year

UNSATURATED

FRACTION

PHA35.000 t(0.7g/g)

Excellent 2nd generationBiofuel!

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The Holistic Nature of ANIMPOLThe 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

Slovenia34

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

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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 ProcessingKey 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!

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• 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

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The Holistic Nature of AnimpolThe Holistic Nature of Animpol

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time [h]

Linear increase of PHA concentration

Biotechnological Example: Fermentation Pattern for PHA Production from Animal-Derived, Saturated Biodiesel

μ max. = 0,20 1/h

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Process Parameters ValuesCell Dry Mass 45,7 [g/L]

PHA 30,2 [g/L]

Residual Biomass 15,4 [g/L]

PHA / CDM 66,2 [%]

µ max. 0,20 [1/h]

Volumetric Productivity 0,62 [g/Lh]

Yield Biomass / Biodiesel 0,6 – 0,7 [g/g]

Main Results:

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1. General Impact: • solutions for waste problems are elaborated on

local scales, but they can be applied for all Europe!

1. Transitional Impact:• creation of ecological and economic benefits by

converting waste into value-added materials

1. Socioeconomic Impact: • new jobs directly in the involved industrial

branches and high-qualified scientific jobs in academia.

The WHEYPOL and the ANIMPOL Project: ImpactsThe WHEYPOL and the ANIMPOL Project: Impacts

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??

June 2008: Price surmounted 130 US-$ per barrel (BRENT)July 2008: Price surmounted 140 US-$ per barrelJanuary 2009: Back to less than 40 US-$ per barrelJune 2010: Again 77 US-$ per barrel!!April 2011: 127 US-$ per barrel!!April 27, 2012: 120 US-$ per barrel!!TOMORROW: WHO KNOWS????? Uncertainties in global political situation!

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1. Utilization of waste materials.

2. Integration into existing production line.

3. Alternative extraction methods.

AcknowledgementsAcknowledgements

The audience and the organizers!

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