TE USE OF CRUDE GLYCEROL AND LIGNOCELLULOSICYDROLYSATES IN TEPRODUCTION OF 1,3 PROPANEDIOL FOR BIOPLASTICS Josef Modl, EVP in co operation with www.vogelbusch.com Vienna University of Natural Resources and Life Science
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Agenda About ourselves the VOGELBUSC Group Worldwide turnaway from fossil energy sources biodiesel Our proven concept crude glycerol to valuable 1,3 propanediol Another waste stream gets new commercial and green values Conclusion Open discussion www.vogelbusch.com
Who is the Vogelbusch Group? Independent Austrian based engineering company Founded in 1921 by Mr. Wilhelm Vogelbusch > 100 highly qualified in house technologists and technicians > 50 patents and proprietary technologies filed Biotechnology projects realized in > 80 countries of the world Long lasting collaboration with university research facilities www.vogelbusch.com
The Vogelbusch Group white biotechnology red biotechnology Vogelbusch Biocommodities Gmb Vogelbusch Biopharma Gmb Vogelbusch U.S.A. Inc. Vogelbusch Anlagenservices Gmb Vogelbusch ong Kong Ltd.
Our key competences white biotechnology Engineering and construction of bioprocess plants for the sugar and starch processing and the food industries. Proprietary technology for alcohol, vinegar, yeast, organic acids, starch sugars. Unique expertise in the key biotechnological tasks of fermentation, separation, distillation, evaporation for industrial design. www.vogelbusch biocommodities.com www.usa.vogelbusch.com
Our core competences red biotechnology Engineering and construction of biotech plants for the pharmaceutical industry based on relevant GMP/FDA guidelines Realization of upstream and downstream processes Design of white (production) and grey (utility) plant areas Design of key equipment Microbial and mammalian production lines Supply of prefabricated process units in own assembling shop including factory acceptance tests www.vogelbusch biopharma.com
The Vogelbusch Group and our Industrial Clients Proprietary alcohol technology by Vogelbusch filled the gas tank with Biofuels; Selected reference customers www.vbusa.com www.vogelbusch.biocommodities.com Proprietary cell culture and fermentation technology by Vogelbusch production serums for vaccanation; Selected reference customers www.vogelbusch.biopharma.com
Scientific and Commercial Background Transesterification Biodiesel Glycerol Re-use of glycerol as substrate for a biorefinery for block-building chemicals?
Biodiesel Glyceryl trioleate Primarily driven by government policies and declining vegetable oil prices biodiesel production will advance in 2013 (forecast) to about 24.7 mln t, which result to about 2.7 mln t crude glycerol produced (averaged).
Biochemical basics O Glycerol dehydratase (dhab1-3) 1,3-Propanediol oxidoreductase (dhat) O O O O O O Glycerol 3-ydroxy Propionaldehyde 1,3-Propanediol Main Known bacterial strains for transformation: Klebsiella pneumoniae Clostridium B. Citrobacter sp. human pathogen
Our workhorse for this conversion process Keyfeatures Non human pathogen GMO for optimal product yields and concentration in mash Easy to cultivate and maintain
Demonstration in small technical plant scale Fed batch culture 50l Scale up bioreactor by Vogelbusch 45 40 aufkonzentrierter Feed 100 90 35 80 30 25 70 60 OD 600 20 15 50 40 30 [g/l] 10 20 5 10 0 0 0 20 40 60 80 100 120 140 160 180 200 time [h] OD600 glucose lactic acid 1,3-PDO acetic acid ethanol glycerol 3-P 0.8 mol 1,3 Propanediol/ mol Glycerol > 80g/L 1,3propanediol
With Cellulosic Sugars gets new Values Sugar is needed to promote bacterial grow 1.4 to. crude gylcerol + 0.25 to. C6 sugars = 1 to. 1,3 Propanediol ====================== Solution ydrolysates from empty fruit bunches
Direct comparison in small technical plant scale* * As reference fermentations 100% C6 Sugar guided process ydrolysates guided process (24% C5/ 76% C6) 30 25 20 15 10 5 0 120 110 100 90 80 70 60 50 40 30 20 10 0 0 20 40 60 80 100 120 140 160 180 200 30 25 20 15 10 5 0 120 110 100 90 80 70 60 50 40 30 20 10 0 0 20 40 60 80 100 120 140 160 180 200 Q&A: Can you see any main difference? A: Slightly less biomass production with similar product yields!
Price Comparison Raw Material 395,00 355,00 315,00 275,00 650.00 600.00 550.00 Source: Milling & Baking News April, 2014 Glucose 80% DS, Wholesale US 500.00 1 2 3 4 5 6 7 8 9 10
Commercial scale process design Sufficient data is now available to offer a solid, commercial scale upstream process design. Simple solutions and low complexity are the key features to ensure low investment and production costs.
Where to go from here? GMO to optimize output Glycerol dehydratase (dhab1-3) 1,3-Propanediol oxidoreductase (dhat) O O Glycerol O O O 3-ydroxy Propionaldehyde O O 1,3-Propanediol psm Genome sequencing Transformation protocol Functional plasmid for plasmid expression
Effective? Expression of propanediol oxidoreductase increases propanediol production
Conclusions L. diolivorans is a non pathogen, well suited bacteria for 1,3 Propanediol production from biodiesel plant derived crude glycerol. It is not inhibited by raw glycerol or high 1,3 propanediol concentrations 90g/L 1,3 Propanediol can easily be achieved by fed batch culture. We developed a protocol for recombinant protein expression in L. diolivorans Conventional process C6 sugars can be fully substituted by cellulosic hydrolysates at a 1:1 scale without loss of product yield. We are ready to offer a complete up stream commercial scale process design package or Licensing
Selected references Pflügl S, Marx, Mattanovich D, Sauer M. 1,3 Propanediol production from glycerol with Lactobacillus diolivorans. Bioresour Technol. 2012; 119:133 40. Sauer M, Marx, Mattanovich D. Microbial production of 1,3 propanediol. Recent Pat Biotechnol. 2008; 2(3):191 7. PCT/EP2012/071812, Fermentation process for producing chemicals. Pflügl S, Marx, Mattanovich D, Sauer M. Genetic Engineering of Lactobacillus diolivorans. FEMS Microbiol Lett. 2013;344:152 8.
www.vogelbusch.com Thank you for your attention www.vogelbusch.com Ing. Josef Modl j.modl@vogelbusch.com Dr. Gottfried-umbert Sodeck gh.sodeck@vogelbusch.com Ing. Josef Modl j.modl@vogelbusch.com Dr. Michael Sauer michael.sauer@boku.ac.at ans Marx hans.marx@boku.ac.at