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Bioenergy-bioproducts. Agenda. Fermentation Xylitol Lignin Products from glycerin, class discussion based on the article. A Historical Perspective. Evidence of fermentation dates to 6,000 BC Egyptians brewed a beer-like substance Mid-1800s  Louis Pasteur

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  • Fermentation
    • Xylitol
  • Lignin
  • Products from glycerin, class discussion based on the article
a historical perspective
A Historical Perspective

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


 Everything’s from Fermentation

biochemical products

Food & Beverages

Soy Sauce, Pickles

Beer & Wine


Ethanol & Butanol



Insulin, HGH


Cellulase, Rennet

Organic Acids

Lactate, Formate, Succinate




Amino Acids

Biochemical Products

the ideal microbe



Minimal Product Inhibition

High Product Tolerance

Easy to Manipulate


Degrade Lignocellulosic Material

Ferment ALL Resulting Sugars

High Rates and Yields

The Ideal Microbe

microbial diversity


Saccharomyces cerevisiae

Zymomonas mobilis

Thermoanaerobacter BGL1L1


Clostridium acetobutylicum

Escherichia coli

C. beijerinkii

Lignocellulose Digestion

Clostridium thermocellum

Microbial Diversity

media feedstock

the ideal process

Simple  Few steps

Fast/short residence time

Actual yield = theoretical yield

High productivity

Recoverable catalyst (the cells)

Easily separable culture

The Ideal Process
bioreactor breakdown





Headspace Composition



Residence Time

Bioreactor Breakdown

  • 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
  • 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
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 xylitol-sweetened 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
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


5-C & 6-C sugars

  • Ethanol
  • Xylitol

How do we do it? Fermentation

Synthetic sugars

or hydrolysate


Sugar, ethanol, xylitol

analysis HPLC


Mixed synthetic sugars

Experimental Yields:

Xylitol: 70%

Ethanol: 84%





ptd 3 in steam exploded hardwood and softwood mixture
PTD3 in steam exploded hardwood and softwood mixture

Experimental Yields:

Xylitol: 68% Ethanol: 100%





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
  • 3-dimensional phenolic polymer
  • Complex structure
  • Composes ~15-40% of lignocellulosic biomass
  • 2nd most abundant natural polymer


lignin current use
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
  • 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
Arboform: chemical properties
  • Pelletized mixture of lignin, fine fibers of wood, hemp or flax, and wax
    • Up to 50% lignin
  • Liquifies at temperatures as low as 170°C
    • 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
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

Some forms are not water resistant

Requires removal of sulfur

Cost: $1.60/lb, compared with less than $1/lb for polypropylene

Arboform: pros and cons
  • Advantages:
  • Completely biodegradable
  • Can be burned after use
  • Not made from crude oil
  • At least as strong as plastic

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 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 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

dye dispersant
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
binding dust control1
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
Pellet binder
  • The natural stickiness of lignosulfonates help them function as a pellet binder; it helps hold the material together.
glycerin and products
Glycerin and products
  • Class discussion

“Bioenergy Lab” on Alcatraz

At the 31st Symposium on Biotechnology for Fuels and Chemicals