Pilot plant for the utilization of agricultural starchy raw materials

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Leibniz-Institut für Agrartechnik Potsdam-Bornim e.v. Max-Eyth-Allee 100, D-14469 Potsdam Fon: 0331/5699-112, E-mail: jvenus@atb-potsdam.de Pilot plant for the utilization of agricultural starchy raw materials Joachim Venus Biorefinica 2009 International Symposium Biobased Products and Biorefineries January 27 and 28, 2009, Osnabrück

Bioconversion technologies for the production of industrial raw materials Biorefineries: factories of the future Faced by global warming and limited fossil resources, and thanks to the development of new biological processes, the use of renewable materials becomes an attractive alternative as feedstock for producing fuels, materials and chemicals. Renewable resources and bioprocesses for a sustainable industrial production

Top Value Added Chemicals from Biomass Volume I Results of Screening for Potential Candidates from Sugars and Synthesis Gas August 2004

New biorefinery-concept for starchcontaining crops and green biomass -BIOCONVERSION-

Production figures and prices for fermentation products World production (ton/year) World market price ( /kg) Bio-ethanol 26.000.000 0,40 L-Glutamic acid (MSG) 1.000.000 1,50 Citric acid 1.000.000 0,80 L-Lysine 350.000 2,00 Lactic acid 250.000 2,00 Vitamin C 80.000 8,00 Gluconic acid 50.000 1,50 Antibiotics (bulk products) 30.000 150,00 Xanthan 20.000 8,00 L-Hydroxyphenylalanine 10.000 10,00 Antibiotics (specialities) 5.000 1.500,00 Dextran 200 80,00 Vitamin B12 10 25.000,00

Occurence and application of lactic acid Lactic acid (2-hydroxypropanoic acid), CH 3 -CHOH-COOH [CAS 50-21-5], is the most widely occurring hydroxycarboxylic acid present in many foods both naturally or as a product of microbial fermentation (e.g. sauerkraut, yogurt, buttermilk, sourdough breads) and many other fermented foods Main fields of application Traditionally, food and food-related applications: acidulant, flavoring, ph buffering agent, inhibitor of bacterial spoilage, emulsifying agents particularly for bakery goods... Non-food: - textile and leather auxiliary, mordant, cleaning agent, disinfectant, ph adjustment of hardening baths... - Lactic acid and ethyl lactate [CAS 97-64-3] have long been used in pharmaceutical and cosmetic applications and formulations (topical ointments, lotions, anti acne solutions, humectants, parenteral solutions and dialysis applications, anti carries agent...) - chemical industry (raw material for the production of lactate ester, propylene glycol, 2,3-pentanedione, propanoic acid, acrylic acid, acetaldehyde, dilactide, biodegradable polymers...)

Overview of the two manufacturing methods of lactic acid Petrochemical resources Acetaldehyde pre-treatment acid hydrolysis and/or enzymatic saccharification Renewable resources simultaneous saccharification and fermentation addition of HCN and catalyst Lactonitrile microbial fermentation Fermentable carbohydrates hydrolysis by sulfuric acid only racemic DL-lactic acid recovery and purification Fermentation broth optically pure L(+)- or D(-)-lactic acid Desirable due to Recent environmental issues Limited nature of petrochemical resources [WEE et al., 2006]

COMMUNICATION FROM THE COMMISSION TO THE COUNCIL, THE EUROPEAN PARLIAMENT, THE EUROPEAN ECONOMIC AND SOCIAL COMMITTEE AND THE COMMITTEE OF THE REGIONS on the mid term review of the Strategy on Life Sciences and Biotechnology Summary of recommendations from the contact network Support the setting up of demonstration/pilot projects and integrated bio-refineries, which are flexible installations at pilot or industrial scale for the production of biofuels and other biomaterials, based on a variety of feedstock. Giving support to demonstration projects is important since SMEs active in this area do not have the resources to set up a real proof-of-concept. It would also help to test logistical solutions and form value chain coalitions between actors COMMISSION OF THE EUROPEAN COMMUNITIES Brussels, 10.4.2007, SEC(2007) 441 Universities, Research Institutes, SMEs Applied & basic research Industry Industrial application Large-scale production

Pilot plant facility swift transfer of new biotechnological processes into practice often fails due to the lack of a reference facility that can be used for multiple applications pilot facility for production of lactic acid at the ATB consequently fills a gap in the various phases of bioprocess engineering provision of product samples is intended to open up the possibility of interesting partners in industry with specific product requirements in the various applications scale up Pilot fermentor Type P, 450 L (Bioengineering AG) BIOSTAT Bplus (Sartorius BBI Systems GmbH, Germany) equipped with a digital control unit DCU for the continuous fermentation with cell recycling Venus, J.; Richter, K.: Eng. Life Sci. 2007, 7, No. 4, 395-402

Process steps for the manufacture of lactic acid Raw material storage 5 m 3 (Trevira UV silo, HIMEL Maschinen GmbH & Co. KG) Hydrolysis Pre-, Microfiltration 1 m³ stirred vessel; 0.5 m³ storage tank (Apparate und Behältertechnik Heldrungen Gmbh) 0.8 mm coarse filter (Sommer & Strassburger GmbH & Co. KG), Microfiltration (ZrO 2 -TiO 2 CeRAM INSIDE, TAMI Industries France) Sterilization of the nutrient broth/additives 2x400 L, 2x250 L stirred vessels (Apparate und Behältertechnik Heldrungen Gmbh) Fermentation with cell retention Softening Pilot fermentor Type P, 450 L (Bioengineering AG) MOLSEP Hollow fibre PES membrane (FS10FUS, MICRODYN-NADIR GmbH/Daicen Membrane Systems Ltd.) Rondomat HW A 250 DWZ, 2x3 m³/h (BWT Wassertechnik GmbH Schriesheim) Monopolare/Bipolare Electrodialysis FT EDR/ ED4 15 (FuMA-Tech GmbH) Ion exchange Cationic resin EXC 08, 50 L; Anionic resin EXA 133, 2x90 L (UIT GmbH Dresden)

Bioconversion of renewables feedstock cereals (e.g. rye), wholemeal green biomass pretreatment bioconversion cereal s hydrolyzate press juice fermentation, down-stream products (raw)lactate, lactic acid, biomass... bioplastics

Enzymatic grain/starch hydrolysis ( C 6H10O5 ) n (n 1) H 2O + n C6H12O 6 Glucose Hydrolysis (1 m³) Separation of wholemeal particles Hydrolyzate of Rye (081104) DM [%] NH 4 -N [mg/l] N ges [g/l] P ges [mg/l] Glucose [g/l] End of hydrolysis 16,5 22,9 3,103 711 103,9 After filtration # 13,5 29,3 1,547 589 101,2 # Enrichment & purification of the hydrolyzate by coarse filtration & membrane separation processes Membrane filtration

Mass flow fermentation unit (batch) Dextrose S (24 kg) ph correcting agent Nutrients, salts etc. Inoculum Ultrafiltration Fermentation Y P S ΔP / = = 0,89 Δ S 268 Litres Filtrate Lactate P (21,3 kg) 38 Litres Biomass (3,0 kg Lactate)

Repeated batch cultivation Lactobacillus paracasei 40,5 C; ph 6,0 Rye hydrolyzate with added nutrients 60 biomass sugar lactate sugar, lactate, biomass [g/l] 40 20 0 0 50 100 150 200 250 time [hours] 300

New continuous fermentation process with cell retention In the present case, a continuous operating mode is used, in which part of the biomass formed is retained in the system. The main portion of the fermented medium leaves the bioreactor as a cellfree filtrate through a membrane module, while a second process (containing biomass) is necessary to maintain a stable steady state. The principle of the fermentation mode is illustrated here.

Screenshot of the flow chart biomass separation

Down-stream processing of raw lactates & lactic acid Ion exchange Electrodialysis Evaporation unit

Effect of several down-streaming steps on the purity of lactic acid Na-Lactate (Fermentation) 4.000 3.600 3.200 down-stream processing (Filtration, Softening, Electrodialysis, Ion exchange, Decolorization, Evaporation) 1000 900 800 ions [mg/l] 2.800 2.400 2.000 1.600 1.200 700 600 500 400 300 lactate [g/l] 800 200 400 100 0 0 Start_Ferm End_Ferm MF Soft EDR EXA133 EXC08 Evap

Dependency of the enantiopurity of lactic acid in the course of purification 1000 100,0 g/l 800 600 400 L-MS D-MS HPLC L/D L/D 99,0 [%] 98,0 97,0 200 96,0 0 End_Ferm MF Soft EDR EXA133 EXC08 Conc 95,0

Key topics of the European biomass research [EU action plan, Brussels, 7.12.2005 - COM(2005) 628 final] The Commission s proposal for the 7th Framework Programme gives a high priority to biomass research: Life sciences and biotechnology for sustainable nonfood products and processes, including the use of biotechnology to improve the productivity, sustainability and composition of biomass raw materials and develop new bio-processes. KBBE-2009-3-7-01: Sustainable Biorefineries Call: FP7-2009-BIOREFINERY Development of advanced biorefineries for sustainable processing of biomass into building blocks for the production of bio-based chemicals, materials, second generation biofuels, power and heat. The biorefineries shall demonstrate their performance, sustainability and feasibility at least at pilot scale in an integrated approach.

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