Utilization of green biomass (grass juice) as nitrogen source for lactic acid fermentation

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1 Leibniz-Institute for Agricultural Engineering and Bioeconomy (ATB) Max-Eyth-Allee 100, Potsdam, GERMANY Fon: +49(331) , Utilization of green biomass (grass juice) as nitrogen source for lactic acid fermentation

2 Our mission: History Our research is aimed at sustainable 1927 Experimental farm of the Agricultural University Berlin intensification. We analyze, model and evaluate 1933 Independent research center on agricultural mechanization bio-economic production systems. We develop and 1952 Central institute of agricultural engineering of East Germany integrate new technologies and management strategies for a knowledge-based, site-specific 1992 Reestablished after the reunification of Germany production of biomass, and its use for food, as biobased materials and fuels - from basic research to Today: Leibniz Institute for Agricultural Engineering and Bioeconomy application. - member of the Leibniz Association

3 Research Program Material and energetic use of biomass Coordination: Dr. Joachim Venus Consideration of the entire value chain System s approach Biomass provision (Cultivation, harvest, storage e.g. short rotation wood, hemp) Material use Chemicals & Materials (Fibers, insulation, biotechnological products) biotechnological Energetic use (Biogas, wood pellets, biochar) Valorization of residues, sidestreams etc.

4 Industrial Biotechnology - Using renewable resources for industry Biobased products and processes from renewable resources not only help preserve the environment and climate, but also make a significant contribution to the structural change from a petrochemical to a biobased industry, with related opportunities for growth and employment. Industrial biotechnology, also known as white biotechnology, is an important driving force in this transition. 2010/2011_en May 2012 March

5 Bioraffinerie ist ein integratives Gesamtkonzept für die Konversion von nachwachsenden Rohstoffen zu Chemikalien, Werkstoffen, Brenn- und Kraftstoffen sowie ggf. zur Erzeugung von Energie (zur Eigennutzung und/oder Auskopplung) als Beitrag zum Nachhaltigen Wirtschaften unter möglichst vollständiger Ausnutzung der Biomasse (BM) [Def. VDI-Richtlinie 6310, 2016]

6 27 November 2017 Map of 224 European biorefineries published by BIC and nova-institute Biorefineries are the heart of the bioeconomy. Here, different types of biomass are fully utilised and transformed into a large variety of chemicals and materials The map distinguishes between Sugar-/starch based biorefineries, producing bioethanol and other chemicals (63), Oil-/fat-based biorefineries biodiesel (64) and Oil-/fatbased biorefineries oleochemistry (54), Wood-based biorefineries (25) excluding those that produce pulp for paper only, Lignocellulose other than wood (5) and finally Biowaste-based biorefineries (13). The prevalence of biorefineries differs considerably between countries. The type of biorefinery is clearly dependent on the locally available biomass. Wood-based biorefineries can be found mainly in Northern Europe and no Sugar-/starch (explicit) based biorefineries mainly in France, Belgium, Germany and Hungary, where we see high yields in sugar and starch. green To create this map, Cologne-based nova-institute conducted a comprehensive survey of all European biorefineries in summer The project was biorefinery done on behalf of the Bio-based Industries Consortium (BIC). For this purpose, a large listed... number of sources were evaluated and an online survey was carried out, during which biorefineries could register themselves. Eventually, so far 224 (!?) biorefineries were identified and mapped across Europe. The map can be downloaded for free at or

7 Building blocks that could be produced via fermentation SpecialChem - Aug marketsandmarkets The market for lactic acid is growing as it is largely used in various industrial applications such as in biodegradable polymers, food & beverages, personal care products, and pharmaceutical industries. The lactic acid market is mainly driven by its enduse industries. In 2013, Biodegradable polymers formed the largest application for lactic acid, followed by food and beverages. The lactic acid market is estimated to grow at a CAGR of 18.8% from 2014 to reach $3,577.5 million by Numbers next to biochemicals designate the total annual production in thousands of t

8 The processes for producing lactic acid from biomass/residues include the following 4 main steps: (1) Pretreatment - breaking down the structure of the feedstock matrix (2) Enzymatic hydrolysis - depolymerizing biopolymers like starch, cellulose etc. to fermentative sugars, such as glucose (C6) and xylose (C5), by means of hydrolytic enzymes (3) Fermentation - metabolizing the sugars to lactic acid, generally by LAB (4) Separation and purification of lactic acid - purification of lactic acid to meet the standards of commercial applications Pilot plant facility for lactic acid fermentation at Leibniz Institute for Agricultural Engineering and Bioeconomy (ATB Potsdam)

9 In der Landwirtschaft sind Grünlandflächen für die Agrarproduktion wichtig, aber auch für den Klimaund Naturschutz sowie die Landschaftsqualität (Ökosystemdienstleistungen). Rohstoffpotenziale für die Grüne Bioraffinerie - Grünland Brandenburg Dauergrünlandfläche seit 1990 stabil Tierbestände auf Grünland seit 1990 um 50% reduziert Grasrationen beim Milchvieh rückläufig Dauergrünland # meist extensiv genutzt Steigerung der Nutzungseffizienz in Dauergrünland möglich # Dauergrünland ist eine landwirtschaftliche Fläche, die gegenwärtig und seit mindestens 5 Jahren zum Anbau von Gras oder anderen Grünfutterpflanzen genutzt wird, auch wenn die Fläche in diesem Zeitraum umgepflügt und eine andere als die zuvor dort angebaute Grünfutterpflanzenart eingesät wird. Amt für Statistik Berlin-Brandenburg Papendiek, F., Ende, H.-P., Steinhardt, U., Wiggering, H., Biorefineries: Relocating Biomass Refineries to the Rural Area. 9

10 M.A. Abdel- Rahman et al. Journal of Biotechnology 156 (2011) Table 1: Overview of chemicals that are currently produced, or could be produced, from biomass together with their respective market type, size of the market, and potential biomass feedstock. Major players involved are also given

11 Chemicals from Biomass: A Market Assessment of Bioproducts with Near-Term Potential Mary J. Biddy, Christopher Scarlata, and Christopher Kinchin - National Renewable Energy Laboratory Data Gaps Scale-up of lactic acid production would require clean, cheap sugars from lignocellulosic biomass to compete with commodity sugar and starch substrates. There is a lack of data about lactic acid production and purification from biomass hydrolysates, including issues of C5 sugar utilization, although it appears work has started to address some of these issues. This report is available at no cost from the National Renewable Energy Laboratory (NREL) at Technical Report NREL/TP March 2016/Contract No. DE-AC36-08GO

12 Example green biomass: Grass processing with a screw press into juice and pellet 18 Lactat Biomasse Glucose [g L -1 ] [g L -1 ] [g L -1 ] 150 Biomass Glucose, Lactate Vodnar, D.C.; Venus, J.; Schneider, R.; Socaciu, C.: Lactic Acid Production by Lactobacillus paracasei 168 in Discontinuous Fermentation Using Lucerne Green Juice as Nutrient Substitute. Chemical Engineering & Technology 33(2010) No. 3, Papendiek, F.; Venus, J.: Cultivation and fractionation of leguminous biomass (open symbols), and green juice (solid symbols) as an for lactic acid production. Chem. Biochem. Eng. Q., 28 (3) (2014) time 50 [hours] Typical time course of a batch lactic acid fermentation supplemented by conventional nutrients alternative source 0

13 Performance of the continuous fermentation based on cereals hydrolyzate Continuous test conditions Biomass [g L -1 ] Lactate [g L -1 ] Productivity [g L -1 h -1 ] synthetic salts & nitrogen sources 9,2 80 7,5 synthetic salts & nitrogen sources 18,0 50 9,1 change to green juice ,5 green juice after coarse filtration ,0 green juice after microfiltration ,3 Reduction of the nitrogen/mineral salt concentration for the fermentation with respect to the following down-stream processing Purification of the raw lactate up to a quality of the commercial available substance Development of a basic continuous lactic acid fermentation process with cell retention and control of the glucose concentration Venus, J.: Utilization of renewables for lactic acid fermentation. Biotechnol. J. 2006, 1, No. 12,

14 Composition of liquid phases in the alfalfa press juice at the Müncheberg site g/l 10 DM 10[%] Glucose Fructose Disacch Nitrogen Dry matter/% Papendiek, F.; Venus, J.: Cultivation and fractionation of leguminous biomass for lactic acid production. Chem. Biochem. Eng. Q., 28 (3) (2014)

15 Lactate [g/l] Substrate [g/l] Product formation (A) and substrate consumption (B) with different nitrogen concentrations at the Müncheberg site g/l N 4.21 g/l N g/l N g/l N 4.21 g/l N 3.68 g/l N time [hours] time 40 [hours] 50 The results for the green juice with the lowest nitrogen content indicate speedier product formation but incomplete sugar consumption There is also a delay in the performance of samples with a higher nitrogen concentration Considering the partially different sugar concentration, lactic acid yield exhibits slightly higher values for experiments conducted using press juice containing higher levels of nitrogen Papendiek, F.; Venus, J.: Cultivation and fractionation of leguminous biomass for lactic acid production. Chem. Biochem. Eng. Q., 28 (3) (2014) 15

16 Dietz, D.; Schneider, R.; Papendiek, F.; Venus, J.: Leguminose green juice as an efficient nutrient for L(+)-lactic acid production. Journal of Biotechnology 236 (2016) 26 34

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18 Mulchfolie aus Poly-Milchsäure ist biologisch abbaubar. F. Kesselring, FKuR Willich Milchsäure für Bio-Plastik und Medizintechnik Biotechnologisch werden Laktobazillen eingesetzt, um im industriellen Maßstab Milchsäure herzu-stellen weltweit etwa Tonnen pro Jahr. Als Lebensmittelzusatzstoff (E 270) erhöht Milchsäure die Haltbarkeit von Back- und Süßwaren sowie Limonaden. Auch Seifen, Cremes und Spülmittel enthalten die desinfizierend wirkende Milchsäure. Durch Verknüpfung mehrerer Milchsäure- Moleküle entstehen Milchsäure-Ketten, die Polylactide. Daraus gewonnene Materialien sind stabil, aber biologisch abbaubar, sodass sie zu Bio-Folien und -Verpackungen verarbeitet werden. Medizintechniker verwenden Polylactide für Nahtmaterialien und Implantate, die sich nach einiger Zeit im Körper zersetzen. Informationen, Experten-Kontakte, Bildmaterial: Dr. Anja Störiko Tel

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