Products of industrial microbiology

Transcription

1 Tortona, 4th April 211 MICROORGANISMS: THE REAL STARS OF THE ENERGY / CHEMICAL PRODUCTION FROM BIOMASS Favaro L., Basaglia M., Casella S. Dipartimento di Biotecnologie Agrarie Products of industrial microbiology The products may be the cells themselves or products made from cells. In the case of bioconversion, cells are used to biochemically convert specific substances. Cells Bioconversion Products from cells Substrate Enzymes (cellulases, ) Chemicals (biopolymer, ) Yeasts cells Product Antibiotics (penicillin, ) Food additives (leucine, ) Fuels (ethanol, ) 1

2 AGRICULTURE FOOD OOD ENVIRONMENT DBA Microbes and biorefineries 2

3 PADOVA s ACTIVITIES ON BIOREFINERY BIOPOLYMERS BIOETHANOL BIOHYDROGEN SUBSTRATE High Cost TRANSFORMATION PRODUCTION of USEFUL compounds Out of market? SUBSTRATE Low Cost - DISPOSAL - ENVIRONMENT TRANSFORMATION RECYCLING EXTRACTION VALORIZATION PRODUCTION of USEFUL compounds Potential market ONE process and MANY utilities 3

4 Alternative substrates for biopolymer production The only food industry annually produces thousand of tons of ORGANIC WASTES which represent huge amounts of low cost C by-products Whey Animal wastes EU-WHEYPOL GRD Dairy industry waste as source for sustainable polymeric material production Technical University of Graz, Institute of Biotechnology, Austria Technical University of Graz, Institut für Thermische Verfahrenstechnik, Austria BDI Anlagenbau Ges.m.b.H. University of Padova, Dipartimento Biotecnologie Agrarie, Italy ALVI, Cooperativa Latte Alto Vicentino S.c.ar.l. University of Pisa, Department of Chemistry & Industrial Chemistry, Italy IDROPLAST Srl Polymer Institute of the Slovak Academy of Sciences Centre of Polymer Chemistry - Polish Academy of Sciences, Poland National Institute of Chemistry, Slovenia Polyhydroxyalkanoates (PHAs) 4

5 EU-ANIMPOL FP7-KBBE-29-3 Biotechnological conversion of carbon containing wastes for eco-efficient production of high added value products Graz University of Technology, Austria University of Padova, Dipartimento Biotecnologie Agrarie, Italy University of Zagreb, Croatia Argent Energy, U.K. Termoplast, Italy University of Pisa, Italy Polish Academy of Sciences, Poland National Institute of Chemistry, Slovenia Argus Umweltbiotechnologie Gmbh, Germany U. Reistenhofer GesmbH, Austria Bioethanol 5

6 Azione Biotech II Direct microbial conversion of cellulose into ethanol Promolog Srl Dipartimento di Biotecnologie Agrarie Università di Padova Veneto Agricoltura PhD School on Crop Sciences Azione Biotech III Optimisation of the energy production processes from bioethanol Dipartimento di Biotecnologie Agrarie Università di Padova Dipartimento di Principi e Impianti di Ingegneria Chimica I. Sorgato, Università di Padova Centro Interuniversitario di Contabilità e Gestione Agraria, Forestale ed Ambientale, Università di Padova Azienda Agricola Ca Corniani, Caorle Cereal Docks spa, Camisano Vicentino Istituto per l Energetica e le Interfasi, CNR, Padova Department of Microbiology, University of Stellenbosch 6

7 Consolidated BioProcessing of agricultural residues Cellulose Lignin Glycosyl Hydrolases Hemicellulose Wheat bran 31% Hemicellulose 28% Starch 11% Cellulose Ethanol + CO 2 Potato peel 1% Hemicellulose 5% Starch 15% Cellulose Laboratory vs wild type strains Laboratory Wild type Genetically defined, haploid Multiple auxotrophic mutations (leu2, ura3, his3, trp1) Suitable for gene cloning Low ethanol yield Low ethanol tolerance Low inhibitor tolerance Genetically unstable, polyploid No auxotrophic mutation (leu2, ura3, his3, trp1) Difficult gene cloning High ethanol yield High ethanol tolerance Good inhibitor tolerance 7

8 Screening of of cellulase and amylase-encoding genes Van Zyl, W.H., Jooste, T., Görgens, J.F., Saayman, M., Favaro, L., Basaglia, M., Casella, S. (21). Method for Engineering Yeast (PCT/IB21/795). pbcf1 δ-integration in S. cerevisiae industrial strains S. cerevisiae Does it interfere with the industrial fitness of the strains? Selection of stable recombinant strains Is the selection of recombinant yeasts sufficient for a successfully CBP process? The case of a glucoamylase gene 8

9 First check: do they ferment glucose? Glucose Ethanol Glucose Ethanol g/l Genes integration does not 15 interfere with fermentative performance 1 of yeast g/l Time (h) Time (h) S. cerevisiae Yield.491 (96%) Yield.487 (~96%) Second check: do they yield glucose from starch? 5 C, 1' 3 C, 3' 5 C, 1' 3 C, 3' 3 15 nkat(g s.s.) nkat(g s.s.) Wild type Mutant Wild type Mutant Soluble starch Raw starch 9

10 Third check: do they ferment starch? S. cerevisiae Starch Ethanol Starch Ethanol Starch (g/l) Ethanol (g/l) 4 2 Starch (g/l) Ethanol (g/l) Time (h) Time (h) No Yield Yield.44 (~79%) A CBP yeast for starchy (and cellulosic) substrates? α-amylase Glucoamylase Yeast Brans Liquefaction Saccharification Fermentation Distillation & dehydration Potato peels Water Jet Cooker >1 ºC 5-8 min Storage tank 6 ºC 8-1 h Slurry tank Grinding Adjust ph to 6. Adjust ph to 4.5 Secondary Liquefaction 95 ºC, ~9 min DDGS (Animal feed) Fuel blending 1

11 A CBP yeast for starchy (and cellulosic) substrates? α-amylase Glucoamylase Yeast Brans Liquefaction Saccharification Fermentation Distillation & dehydration Potato peels Water Jet Cooker >1 ºC 5-8 min Storage tank 6 ºC 8-1 h Slurry tank Grinding Adjust ph to 6. Adjust ph to 4.5 Secondary Liquefaction 95 ºC, ~9 min DDGS (Animal feed) Fuel blending A CBP yeast for starchy (and cellulosic) substrates? Brans Fermentation Distillation & dehydration Potato peels Water Amylolytic yeast Storage tank No enzyme costs Slurry tank Low input Limited nutrients Grinding Adjust ph to 6. DDGS (Animal feed) Fuel blending 11

12 Tortona, 4th April 211 MICROORGANISMS: THE REAL STARS OF THE ENERGY / CHEMICAL PRODUCTION FROM BIOMASS Thank you for your attention Dipartimento di Biotecnologie Agrarie 12