The ANIMPOL Project: From Animal Waste to PHA-Bioplastics

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The ANIMPOL Project: From Animal Waste to PHA-Bioplastics Martin Koller, Anna Salerno, Alexander Muhr, Angelika Reiterer, Heidemarie Malli, Karin Malli, Gerhart Braunegg Graz University of Technology, Austria Institute of Biotechnology and Biochemical Engineering October 24 th to 25 th, 2011, Bologna

Content of the Presentation Objectives and Significance of ANIMPOL The Plastic Situation today PHA Biopolyesters as a sustainable solution Potential Applications of PHAs The Structure of the ANIMPOL Project The Project Consortium Industrial Involvement in ANIMPOL Raw Materials Available for the Process Example of the Outcomings Expected Final Outcomings and Outlook

Objectives of ANIMPOL The project ANIMPOL (»Biotechnological conversion of carbon containing wastes for ecoefficient production of high added value products«) utilises: waste streams from slaughterhouses, the animal rendering industry and waste fractions from conventional biodiesel manufacture for the production of improved biodiesel (fatty acid esters, FAE) and high-value biodegradable polymeric materials (polyhydroxyalkanoates, PHA).

Nowadays, we live in the Plastic Age 100 million tons (total) 250 million tons (only fossil resources) 1,5 million tons (total) 60 years ago 20 years ago 2010

Quantities of Utilized Plastic Materials in Different Global Regions 250 Mtons / a World production & consumption of Plastic Materials 80-120 kg / a Developed Countries (average consumption per person) 2-15 kg / a Emerging and Developing Countries (average consumption per person)

TODAY: Polymers Predominately Deriving from Petro- Industry Highly Resistant Polymeric Materials No natural degradation Insufficient performance of recycling systems High risk connected to the thermal conversion of plastic by inceneration.

It is time to switch... 1. Fluctuation of crude oil 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. The degradation products of these materials contribute to the green house effect and global warming.

PHAs: a sustainable solution! Polyhydroxyalkanoates (PHAs) are biopolymers produced by a broad range of prokaryotes from renewable resources. The industrial implementation has a two major impacts: in replacing petrol based plastics; in solving industrial waste problems.

PHAs: a Sustainable Solution! 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

What characterizes a GREEN Plastic?

When Plastics are GREEN? Biobased Biodegradable Compostable Biocompatible The production of the building blocks is based on renewable resources; the polymerization of the monomers may occur chemically or biotechnologically. The 90% of the carbon of the plastic is metabolized within 180 days. (standardized norm EN-13432) 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)

PHAs: a Sustainable Solution! 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

White Biotechnology Raw materials (hydrolysis) Accessible C source (through fermentative process) Microorganisms (Archea, Bacteria, Fungi) Haloferax mediterranei Xanthomonas campestris (separation and purification) PHAs

PHAs: a Reserve Compound PHAs provide an advantage for microbial surviving! PHAs serve as a storage materials for carbon and energy for the microorganism: produced under conditions of carbon surplus together with a limitation of an essential growth component. metabolised under condition of starvation, this reserve compound, into the final products: H2O and CO2. 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, 218-226, 2007

PHAs: a Sustainable Solution! 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

Potential Applications of PHAs Agro-Industrial Therapeutic Use of Chiral building blocks Packaging Materials Surgical carriers and matrices for controlled release of nutrients, fertilizers and pesticides; mulch foils etc. controlled release of active pharmaceutical ingredients as synthons for synthesis of organic fine chemicals compostable after utilization implants

Application of PHAs Surgical Applications: Implants Ongoing PROJECT BRIC [Laura Bassi Center of Expertises; Austrian project]: Development of BioResorbable Implants for Children surgery (frenum healing). Coordinated by Medical University Graz, Austria; Prof. A. Weinberg Artifical organs, artifical blood vessels, materials for wound treatment: Chen et al., 2005 Sodian et al., 2000 Rokkanen et al., 2000

Obstacles in the Market Penetration of PHAs The production costs of PHA must be in the same range as the competing classical plastics (PP; LDPE) These costs have to be minimized dispite the instable market price for crude mineral oil by optimizing: 1. The selection of raw materials 2. The cost of downstream processing for isolation of PHA from biomass

Waste Streams Selection for Carbon Sources Location of Production Plant Wastestreams available Biopolymer Production integrated into production line No interference with food- or feed applications!!! Biopolymer production based on Renewable Resources

Our Choices... Alternative Carbon Sources: 1. Whey from dairy industry (Lactose): EU-FP6 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) 4. Animal Derived Waste Lipids (EU-FP7 PROJECT ANIMPOL)

FP7 Project ANIMPOL The Animpol project aims at the sustainable and value added conversion of waste-lipids from animals in order to create a viable strategy that enables the production of PHAs in Europe in future.

WASTE LIPIDS Transesterification MIX BIODIESEL-GLYCEROL Separation BIOFUEL (FME) GLYCEROL LIQUID PHASE (GLP) MICROBIAL PHA PRODUCTION (group 1 and group 2 production strains) Downstream Processing EXTRACTION OF PHA FROM BIOMASS Purification/Refining PHA Hydrolysis RESIDUAL BIOMASS Proteins Lipids Waste Fraction

FP7 Project ANIMPOL Biotechnological conversion of carbon containing wastes for eco-efficient production of high added value products Project Start: January 1 st, 2010 Entire Project Volume: 3,7 Mio.; EU contribution: 2,9 Mio Coordinated by Graz University of Technology, Austria

The Holistic Nature 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 Graz University of Technology Università di Padova University of Zagreb Partner Logo Key Researcher Main Roles Country 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; Engineering (Institute of process and Particle Engineering) Austria Prof. Sergio Casella Microbiology, Genetics Italy Prof. Predrag Horvat Mathematical Modeling of Bioprocesses Croatia University of Graz Prof. Martin Mittelbach Enhanced transesterification of animal waste lipids; assessment of composition and quality of raw materials Università di Pisa Prof. Emo Chiellini Characterization of PHAs; formulation of PHAbased composites and blends Polish Academy of Science National Institute of Chemistry Prof. Marek Kowalczuk Dr. Andrej Kržan Characterization of PHA and derived composites and blends Characterization of PHA and derived composites and blends Austria Italy Poland Slovenia

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 (highly saturated biodiesel fractions) producer from tallow and waste cooking oil; delivers saturated biodiesel fraction and crude glycerol phase Key representative: Dr. Mike Scott

Addition Industrial Partners: How industry can support and optimize academic research! 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

Advisory Board of Companies Acting as an Enduser Group Advisory Board members are no beneficiaries of the project; they give advice in how to proceed with the activities 1. Novamont, Italy: biodegradables 2. ChemTex Italia (gruppo Mossi & Ghissolfi; Italy): biobased products 3. KRKA, Slovenia: large scale fermentations

Major Goals Development of an integrated, sound process! industrial Bring together waste producers from animal processing industry and biofuel industry with the polymer industry.

The Holistic Nature of Animpol Biotechnology and Fermentation Technology Dissemination and Exploitation of Results; Marketing of Final Product Microbiology and Genetic Engineering Life Cycle Assessment Chemistry and Chemical Engineering Polymer Chemistry and Polymer Processing

Structure of the Project

Amounts of Waste in EU Significant for ANIMPOL ANIMAL WASTE LIPIDS 500.000 t/y CRUDE GLYCEROL 265.000 metric tons/year BIODIESEL SATURATED UNSATURATED CATALLYTICALLY ACTIVE BIOMASS (0.4-0.5g/g) PHA 120.000 t (0.3g/g) FRACTION 50.000 t/year PHA 35.000 t (0.7g/g) FRACTION Excellent Biofuel!

N limitation! Biotechnological Example: Fermentation Pattern for PHA Production from Animal-derived, Saturated Biodiesel Linear increase of PHA concentration μ max. = 0,20 1/h time [h]

Main Results: Process Parameters Cell Dry Mass PHA Residual Biomass Values 45,7 [g/l] 30,2 [g/l] 15,4 [g/l] PHA / CDM 66,2 [%] µ max. 0,20 [1/h] Volumetric Productivity Yield Biomass / Biodiesel 0,62 [g/lh] 0,6 0,7 [g/g]

Impact of ANIMPOL Project 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 involved industrial branches and high-qualified scientific jobs in academia.

PHA Production: Economical in Future or not? Designing ecologically and economically feasible biopolymers production process: 1. Utilization of waste materials. Thank you for your attention! 2. Integration into existing production line. 3. Alternative extraction methods.