Bioconversion of Distillers Grains to Enhance Utilization

2005 Annual World Ethanol Production~ 46 billion L (all grades)Richt, RFAEnough to Displace ONLY 2% World GasolineConsumption

Total World Ethanol Production for2006 = 8.7 billion gal(33 billion L)46% North America47% BrazilWorld demand for enzymes isexpected to rise 6.5 per centannuallyto nearly $5.1 billion in 2009,according to a new report.

Canada Bio-Ethanol• Terra Grain Fuels building a 0.150 bill L/yrwheat-fed ethanol plant, Saskatchewan• St Clair Ethanol (Suncor) now largest plantin Canada (Jul 2006, 0.200 bill L/yr)• Applied for expansion permits to double• Collingwood Ethanol: 0.050 bill L/yr – wetmill-June 2007• Mid-2008:Riverstone/Carlyle/DominionEnergy ~ 0.380 bill L/yr ethanol & 0.380 billL/yr bio diesel in AlbertaTotal 2.1 bill L/yr by 2008 in Canada

Bush Touring a DuPont Research Lab• President Bush's goal - reduce gasolineusage by 20% in 10 years through:• alternative fuels & vehicle fuel-economyeconomy• Means expansion of US fuel ethanol to 133bill L/yr by 2017) -currently ~ 36 bill L/yr• Is there enough US corn for Bush’s s ethanol plan?

Well . . . . .• ‘The Great Corn Rush’ wasalready sweeping theMidwest before the WhiteHouse set its New AmbitiousTargets• Corn growers say “U.S. corn can, at most,supply just 57 bill L/yr by 2015”• "Nobody believes you can hit that kind ofvolume from grain alone in 10 years," said BobDinneen, , president of the Renewable FuelsAssociation

Corn Dry-Milling Process Overview20 mill bu CornCorn CleaningHammermillDistillationAlpha-Amylase Enzyme(0.02%)Mix Slurry Premix40-60˚CCO 2 293 kgheatFermentation32˚CCooker 90-120˚CYeast and GlucoamylaseEnzymeMashCoolerBack-setcoolAlpha-Amylase Enzyme(0.04-0.06%)Liquefaction 90˚CSaccharification60˚CGlucoamylaseEnzyme(0.06-0.12%)Ethanol 200mill L/yrWholestillageCoarse solidsCentrifugethin stillageEvaporatorsolublessolublesDistiller’s WetGrainsRotary DryerDistiller’sDry Grainsw/Solubles160,000 MTCond.Distiller’sSolubles

Most Controversial of all. . . .• Sudden and massive switch of corn cropsfrom Food to Fuel• Major distortions in world agriculturalmarkets• The US produces about 40% of global cornproduction (70% of world total exports)• Ethanol consumed 6% US corn harvest in2000• Now ~20% of the US corn harvest• Analysts are predicting by next year, roughly half ofthe entire US crop will go to ethanol distilleries

Distiller’s Grains Drying Process-Rotary Dryers• Takes ~1/3total ethanolprocessenergy to drywet grain co-product• Leads tonutritionalDamage• Hugeoperatingcosts

Value-Added DWGS• Abundant & cheap raw material• High nutritional value to start(fat(11%), crude fiber(8%), protein(30%)• Environmental issues resolved• No competition with bio-fuels & human food• Create new products/markets (mono-gastrics)• Capture a sizable increase share of existingmarkets (ruminants)• Quality controlled process

What is Solid State Fermentation (SSF)• Growth of microorganisms on moist solidparticles• Spaces between particles contain a continuousgas phase with minimal visible water• Majority of water is discontinuous and absorbedby solid• 4,000 years old• Considered for enzyme production in 1900s andfor penicillin in the 1940s• Submerged Liquid Fermentation dominated• Revival of interest began in 1970s• Theoretical base for SSF technology began to beestablished in the 1990s

Solid State Fermentation (SSF)Microbes:• Produceenzymes• Convert fiber• Consumenutrients• Altering pH• Release heat• ConsumeoxygenD.A. Mitchell, 2006

Silage - SSF• Preserve damphay• Lactic acidbacteria producelactic acid -lowers the pH -stops any ‘nasty’bacteria fromspoiling it• LAB do notproduce toxic by-products

Fermented Foods - SSF• 4,000 years old• Rhizopus oligosporous• Transforms cookedsoybeans into the highlyregarded tempeh• Mycelium of the fungigrows through thesoybean paste holding ittogether – makes sugars• Characteristic chewytexture, whilefermentation gives it adistinctive flavor

Koji - SSF• Steamed rice that has hadkoji-kin, kin, or koji fungispores, cultivated onto it(Aspergillusoryzae)• Creates several enzymes asit propagates• Break the starches in riceinto sugars• Can be further fermentedby the yeast to make sake

Enzymes - SSFProduction of enzymes usingSSF:• Food• Pulp & paper• Detergents• Ethanol• Bioremediation• Textiles• Pharmaceuticals• Cosmetics• Natural Health Products

Durand Bioreactor at National Institute ofAgronomic Research in Dijon France (50L)Durand, 2003

SSF Technology - Advantages• Mechanically/structurally simple, littlepretreatment, less processing energy, lowwater usage, low water activity reduces riskof contamination• Some SSF products have more desirableproperties – i.e. spore-basedbiopesticideswith more resistance to adverse conditions• Certain enzymes are only induced in SSF• Production of metabolite can be muchhigher in concentration in SSF• Usually a different profile of metabolites inSSF

SSF Technological ChallengesSolid State Bioreactor Design• Morphology of fungi sensitive to shear &mixing• Sensor and control technology underdeveloped• Difficult to model – heterogenity atmicroscale• Particle behavior with fungal hyphaebridging gaps• Mass gas transfer is in voids betweenparticles• Dessication can occur fast• Little research into engineering to overcomegradients in heat, pH, moisture and scale-up

SSF Commercialization Examples• Biocon, , India• Food and speciality enzymes began in 1990• Demonstrated production of lovastatin(antihypercholesteremicagent), mycophenolic acid(an immunosuppressant), Trichoderma viride spores(biocontrol), fungal proteases (food industry)• Alltech, , US & Mexico• SSF for enzymes - $10-million facility - 200 staff35,000 square-foot plant – Serdan Mexico• Construction of a $17.7 million SSF plant inKentucky• Allzyme® from locally grown wheat bran (phytase(byAspergillus niger)

In Summary – BTR Bio-Refining Technology:• Eliminate current drying process to reducenutritional damage & energy utilization• Create value-added added nutritional feed fromDDGS• Control & test for consistency & quality• Apply mycotoxin treatment• No need for enzyme addition – already done• Reduce environmental issues• Alter process to target nutritional needs• Platform technology for enzymes, NHP, etc.• Used on other agricultural residues

In Summary – BTR Bio-Refining Technology:• Help to stabilize the cost of raw materialsfor the feed industry• Create sustainability for the ethanolindustry• Remove bottlenecks in future animalfarming• Increase revenues in Ontario (beef, poultry,pork, milk)• Free up raw materials for energy and food• Increase employment and investmentdollars in Ontario

BTR Laboratories at The Stiller Centre London Ontario. Canada