Bioenergy-bioproducts Agenda  Fermentation » Xylitol Lignin  Products from glycerin, class discussion based on the article   Evidence of fermentation dates to 6,000 BC » Egyptians brewed a beer-like substance  Mid-1800s Louis Pasteur  Late 19th - ~1940  Alcohol Fuels  WWI  Acetone for nitrocellulose  WWII  Penicillin Production  1970s Oil Embargo  Brazil EtOH programs  Today  Everything’s from Fermentation  Food & Beverages » Soy Sauce, Pickles » Beer & Wine  Fuels » Ethanol & Butanol » Methane  Pharmaceuticals » Insulin, HGH  Enzymes » Cellulase, Rennet  Organic Acids » Lactate, Formate, Succinate  Solvents » Acetone   Vitamins Amino Acids  Fast-Growing  Robust  Minimal Product Inhibition  High Product Tolerance  Easy to Manipulate  Safe  Degrade Lignocellulosic Material  Ferment ALL Resulting Sugars » High Rates and Yields  Ethanol » Saccharomyces cerevisiae » Zymomonas mobilis » Thermoanaerobacter BGL1L1  Butanol » Clostridium acetobutylicum » Escherichia coli » C. beijerinkii  Lignocellulose Digestion » Clostridium thermocellum Carbon Sources Vitamins & Growth Factors Glucose Sucrose Glycerol Starch Maltodextrin Corn Sugar, Starch, Cellulose, Sugarcane, Sugar Beet Molasses Antifoaming agents Esters, Fatty Acids, Silicones, Sulphonates, PEG Buffers Calcium Carbonate, Phosphates Lactose Milk Whey Growth Factors Thiamine, Biotin Fats Vegetable Oils Nitrogen Sources Inducers IPTG, Pectin Protein Chelators EDTA, Citric Acid Antibiotics Ampicillin, Kanamycin Inhibitors Rifamycin, Sodium Barbital Trace Elements Cu, Fe, Mn, Mo, Co Ammonia Nitrate Soybean Meal, Cornsteep Liquor, Distillers’ Solubles Pure Ammonia, Ammonia Salts, Urea Nitrate Salts http://en.wikipedia.org/wiki/Industrial_fermentation  Few steps  Fast/short residence time  Actual yield = theoretical yield  High productivity  Recoverable catalyst (the cells)  Easily separable culture  Simple  Controls » pH » Temperature » Agitation » Headspace Composition » Pressure » Volume » Residence Time Bioethanol production and chewing gum? Problems   Baker’s cannot utilize five carbon sugars to produce ethanol Genetically modified microorganisms (E.coli KO11, Z.mobilis, P.stipitis) » have diverse nutrient requirements » not as robust as baker’s yeast » cannot tolerate/metabolize inhibitors generated during pretreatment  For biomass to ethanol process to be economically feasible it has to produce high value co-products Inhibitors  5 groups of inhibitors » Released during pretreatment and hydrolysis – Acetic acid and extractives » By-products of pretreatment and hydrolysis – HMFs and furfurals, formic acid » Lignin degradation products – Aromatic compounds » Fermentation products – Ethanol, acetic acid, glycerol, lactic acid » Metals released from equipment Xylitol (1)  Sweetener » as sweet as sucrose » 40% less calories (suitable for diabetics, does not use insulin to be metabolized)  Recommended for oral health » teeth hardening » antimicrobial properties (causes bacteria to lose the ability to adhere to the tooth stunting the cavity causing process) Xylitol (2)   Feels and tastes exactly like sugar and leaves no unpleasant aftertaste Currently produced via chemical way » acid hydrolysis » hydrogenation and purification (expensive)  Uses: » natural sweetener » chewing gun » tooth paste Xylitol (3) Imagine eating guilt xylitolsweetened brownies or knowing that xylitol-sweetened chewing gum is preventing cavities and gum disease. With xylitol, you can now have your sweet tooth and treat it, too! Pink yeast    Novel, naturally occurring, robust yeast from genus Rhodotorula selected from poplar trees Tolerant of (and capable of metabolizing) high concentrations of fermentation inhibitors Rapidly and effectively utilizes both hexose and pentose sugars PTD3 5-C & 6-C sugars Ethanol Xylitol How do we do it? Fermentation Synthetic sugars or hydrolysate Flasks Sugar, ethanol, xylitol analysis HPLC Mixed synthetic sugars Experimental Yields: Xylitol: 70% Ethanol: 84% 6C 5C EtOH XOH PTD3 in steam exploded hardwood and softwood mixture Experimental Yields: Xylitol: 68% Ethanol: 100% 5C EtOH XOH 6C Xylitol-conclusions  Yes, Pink yeast is able to efficiently utilize 5 and 6C to produce lots of ethanol and xylitol » » » » » Glucose 85% EOH Galactose 86% EOH Mannose 94% EOH Xylose 64% XOH Arabionse 29% XOH Lignin • 3-dimensional phenolic polymer • Complex structure • Composes ~15-40% of lignocellulosic biomass • 2nd most abundant natural polymer Sakakibara Lignin: current use • • • • In Kraft pulping, lignin is recovered in black liquor 50 million metric tons produced annually worldwide ~95% of this is incinerated for thermal electrical energy Burning generates an average fuel value of 23.4 MJ/kg Arboform • • • • • A lignin-based thermoplastic Made from a mixture of lignin, plant fibers, and waxes Developed by German company Tecnaro in 1998 Appearance and some physical properties similar to wood Moldable like plastic Arboform: chemical properties • Pelletized mixture of lignin, fine fibers of wood, hemp or flax, and wax – • Liquifies at temperatures as low as 170°C – – – • • Up to 50% lignin Polypropylene: ~160°C Polyethylene: 105-120°C Polystyrene: ~240°C Thermally stable up to 105°C Can be injection molded similar to conventional plastic Arboform: physical properties • • Better molding capabilities than plastic Irregular fiber orientation resists warping – • • Flooring & building material Good acoustic properties (speakers & musical instruments) Currently 300 metric tons produced annually Arboform: pros and cons  • • • • Advantages: Completely biodegradable  Disadvantages: • Some forms are not water resistant Can be burned after use Not made from crude oil At least as strong as plastic • Requires removal of sulfur Cost: $1.60/lb, compared with less than $1/lb for polypropylene • An alternative to plastic? • • More than 100 million metric tons of plastics originating from crude oil are produced annually (worldwide) The pacific trash vortex is twice the size of Texas, reaches 300 feet below sea level, and 90% of it is plastic Lignosulfonates  Lignosulfonates is the name for a product containing sulfonated lignin and other wood chemicals. » Mainly from the acid sulfite process. » A small amount from sulfonated kraft lignin.  Before becoming lignosulfonates (marketable product), this material (spent sulfite liquor) is “cleaned up”. » Pulping chemicals are removed. » Sometimes non lignin compounds (sugars, etc) are removed chemically, biologically, or through physical methods. » Often the lignin is chemically modified. » Product is concentrated to a molasses thickness product or to a powder. Lignosulfonates-uses  Dispersant » Concrete, Dyes, Gypsum wallboard  Binder » Road dust control, animal feed  Emulsifier (think an oil and vinegar salad dressing). » Emulsions are finely dispersed drops of oil or wax in water. » Lignin acts a s stabilizer in the emulsion.  Chelating agent » Oil Well Drilling Fluids, Micronutrient Fertilizers  Raw material for chemical production » Vanillin (softwood) Concrete dispersant    Concrete is made up of 3 ingredients: cement, sand, and aggregate. Water is mixed in to make a workable slurry and to harden the concrete. By using a dispersant like lignosulfonates, less water can be used to get the same viscosity slurry. This makes stronger concrete. Image borrowed from JimRadfprd.com Dye dispersant   Dyes used to dye cloth are water insoluble. In order to dye cloth, dye particles are dispersed in water. What this means are the dye particles are small enough that they pretty much act like they are dissolved. A dispersant keep them apart so they don’t get big and sink. Sulfonated lignins do this very well. After dying, the lignin is washed out. Binding-dust control  Dusty roads are considered a health hazard by the government and thus dust control is mandated  Dust can be controlled with water, lignosulfonates or calcium chloride. Binding-dust control   Lignosulfonates cause the particles to pack closer together and also to adhere. This process forms a dust “free” and also more stable road. Pellet binder  The natural stickiness of lignosulfonates help them function as a pellet binder; it helps hold the material together. Glycerin and products  Class discussion Acknowledgements “Bioenergy Lab” on Alcatraz At the 31st Symposium on Biotechnology for Fuels and Chemicals