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ETHANOL
PRODUCTON
Anaerobic Fermentation of alcohol
 Ethanol is used as
solvent
chemical intermediate
fuel
Solvent:-
Resins, pharmaceuticals , cosmetics ,household
cleaning products, industrial solvent
Fuel:-
Used as a bio fuel , In internal combustion engines
Chemical intermediate :-
• petroleum derived chemicals
• Butadiene production
Organisms for ethanol production:-
Bacteria:-
Many bacteria can produce ethanol
Many produce end products
• Other alcohol
• organic acids
• Polyols
• Ketones gasses
Many bac metabolize glucose by EMP pathway
• Clostridium sps
• Spirochaeta sps
• Sarcina sps
• Streptococcus sps
Yeast
Primary interest are saccharomyces sps
Organisms are selected by
• High growth and fermentation rate
• High ethanol yield
• Osmotolerance
• Low PH
• High temp
• Hardiness under physical and chemical stress
• Ethanol and glucose tolerance(allows conversion of
conc feeds to conc products)
RAW MATERIALS
Saccharine
Starch &
Cellulose materials
SACCHARINE:-
Simple, directly fermentable, 6 & 12 C molecules
Glucose, fructose, maltose
FRUITS:-
Grapes – 15%
Banana – 13.8%
Apples – 12.2%
Pineapples – 11.7%
Pears – 10%
Peaches – 7.6%
Oranges – 5.4%
Watermelon – 2.5%
MOLASSES:-
Beet or cane molasses
Contain 52 – 55 % fermentable
sugars
Molasses with above 15 – 20 %
sugar cont will need to be dilute
A ton yeild-70-80 gallons
CANE SORGUM :-
14% fermentable sugar content
Extraction requires heavy duty
13-14 gallons of alcohol / ton
Acidification is necessary
SUGAR BEET :-
Mostly preferred
15% sugar, 82% water, small amount of starch
Juice Is extracted by crushing
A ton yield 20 – 25 gallons of alcohol
Enzymes improve the alcohol yield
SUGAR CORN WASTE :-
7 – 15% sugar
Sugar is extracted as same as sorghum
8 – 18% gallons of alcohol per ton
Back slopping in the range of 20-25%
Acidification is necessary
Sugar beet sugar corn
Starchy materials:-
Materials that contain more complex carbohydrates
Grains :-
Grains must be milled, diluted & coked & fermented
Contain large amounts of fermentable materials
Alcohol yield is dependent on converted starch
Jerusalem artichokes :-
Abundant source of alcohol ( 16 – 18%) fermentable
material
A ton should yield about 25 gallons of alcohol
Material should be crushed to pulp & cooked for 2-
3 hrs, dilution is not necessary ( 79 – 80% water)
grains artichokes
potatoes sweet potato
PH is adjusted & fermentation is carried out
Potato:-
15 -18% fermentable sugar
A ton yield 20-25% of alcohol
Cooked with steam under pressure
After cooking mash is cooled to conversion for 15-20
min
Once converted PH should be adjusted mash is
fermented
Sweet potato:-
27-28% fermentable sugar
A ton should yield 40 gallons of alcohol
Similar manner to potatoes
Dilution is required (66%water)
Processing steps
Starchy materials are of 2 categories
1. Encased by grain hulls
2. Starch is readily available
Malting:-
Seeds are moistened & allowed to sprout
These seeds are powdered & the powder is called
malt
Milling:-
Grains are milled to expose the starch
Material is ground as fine as possible
But the fine flour is difficult to remove in distillation
process . Pot stills may reduce the problem
Cooking (preparation of mash):-
Starch is liquefied by boiling under pressure
To dissolve the water soluble starches
Always done with steam under pressure at 100°c
Use less water during cooking & water may be
added to dilute the mash
So cooling time may be saved
New method :- Milling & cooking in one operation
without use of water
 The cooked malt is called mash
Conversion :-
Conversion of starch to fermentable sugar (Maltose)
by certain enzymes in the malt or by acid hydrolysis
Enzyme extracts:-
Commercially available
3 basic types :
1. α amylase - produce dextrose
2. Β amylase - produce maltose
3. Gluco amylase – reduce the remaining starch
Acid hydrolysis:-
Conversion by the action of acid
Requires acid proof equipment and high temp
Dilute mineral acid(H₂SO₄) is added to grain slurry
Then immediately neutralized with calcium
hydroxide
Mash cooling:-
Mash is cooled after conversion & PH is checked
Cooling coils are used for cooling
Cooling time is reduced as much as possible
Fermentation
Batch fermentation is commonly employed.
Fermenter volume -- 600mᶟ.
3% inoculum(3×10⁶/ml)
With in 12 days yeast produce 10% ethanol
 when the process carried out at 35- 38°c , ph.4-4.5
max productivity was 1.9 g/h.
80%cells were removed in separator and brought
back again into fermenter.
When the high quality molasses is used, the max
yield is 95%.
Continuous fermentation not commonly used
In US 10 large scale plants among them 4 are used
con fermentation
Nutrient solution was molasses with di ammonium
phosphate
PH was adjusted to 5 with H₂SO₄
Fermentation temp -35°c
After fermentation cells were separated by
centrifugation and channeled back into the first
fermenter
PRODUCT RECOVERY
Cell biomass is separated by centrifugation or
sedimentation.
M.o desired end product – heated ( aggregate)
readily separated from broth by sedimentation.
Desired end product metabolite.
intracellular - n.a , vit ,enz, a.b (gresofulvin)
extracellular - a.a, acitic acid, citric acid, alcohol,
[ penicillin, streptomycin]
Separation of cell biomass
Flocculation flotation
Filtration centrifugation
flocculation:-
Flocculating agent is added
hydrocolloids
inorganic salts
organic poly electrolyte
 produce large aggregate which still settle more
readily
Flotation :-
by introducing gas into liquid
Cells gets adsorbed to the gas bubbles & rise to the
foam layer at the top of vessel
That can be collected & removed from the
bioreactor
Filtration:-
Bacterial cultures – absolute filters
 filamentous Fungi- depth filters
static & cross flow filtration
Distillation
Separation for a mixture of liquids
It relies on differences in B.P of component liquids
to be separated
The mixture to be separated is added to the
distilled pot & is heated
Low B.P will vaporize first. this vapors passes into
distilling head & then into condenser.
With in the condenser the vapor is cooled & it
liquefies.
The resulting liquid is then collected in receiving
flask
Ethanol B.P - 78.5°C
Water B.P - 100°C
Purification:-
By chromatography
HPLC
GLC
Applications
solvent in the manufacture of varnishes and
perfumes
Preservative for biological specimens
In the preparation of essence and flavorings
In many medicines and drugs.
As a disinfectant and in tinctures.
Used as a biofuel (99.2%).
Medically ethanol is soporific.
Used as antiseptic.
Fluid in many alcohol thermometer.
92.4%ethanol is used as solvent in cosmetics
pharmaceuticals and chemical industry
Byproduct
DDGS (distillers dried grain with solubles)
Co₂– used for the production of dry ice and preparation
of soft drinks
Yeast biomass- animal fodder (contains proteins &vit)
Ethanol production
Ethanol production

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Ethanol production

  • 3.  Ethanol is used as solvent chemical intermediate fuel Solvent:- Resins, pharmaceuticals , cosmetics ,household cleaning products, industrial solvent Fuel:- Used as a bio fuel , In internal combustion engines Chemical intermediate :- • petroleum derived chemicals • Butadiene production
  • 4. Organisms for ethanol production:- Bacteria:- Many bacteria can produce ethanol Many produce end products • Other alcohol • organic acids • Polyols • Ketones gasses Many bac metabolize glucose by EMP pathway • Clostridium sps • Spirochaeta sps • Sarcina sps • Streptococcus sps
  • 5. Yeast Primary interest are saccharomyces sps Organisms are selected by • High growth and fermentation rate • High ethanol yield • Osmotolerance • Low PH • High temp • Hardiness under physical and chemical stress • Ethanol and glucose tolerance(allows conversion of conc feeds to conc products)
  • 7. SACCHARINE:- Simple, directly fermentable, 6 & 12 C molecules Glucose, fructose, maltose FRUITS:- Grapes – 15% Banana – 13.8% Apples – 12.2% Pineapples – 11.7% Pears – 10% Peaches – 7.6% Oranges – 5.4% Watermelon – 2.5%
  • 8. MOLASSES:- Beet or cane molasses Contain 52 – 55 % fermentable sugars Molasses with above 15 – 20 % sugar cont will need to be dilute A ton yeild-70-80 gallons CANE SORGUM :- 14% fermentable sugar content Extraction requires heavy duty 13-14 gallons of alcohol / ton Acidification is necessary
  • 9. SUGAR BEET :- Mostly preferred 15% sugar, 82% water, small amount of starch Juice Is extracted by crushing A ton yield 20 – 25 gallons of alcohol Enzymes improve the alcohol yield SUGAR CORN WASTE :- 7 – 15% sugar Sugar is extracted as same as sorghum 8 – 18% gallons of alcohol per ton Back slopping in the range of 20-25% Acidification is necessary
  • 11. Starchy materials:- Materials that contain more complex carbohydrates Grains :- Grains must be milled, diluted & coked & fermented Contain large amounts of fermentable materials Alcohol yield is dependent on converted starch Jerusalem artichokes :- Abundant source of alcohol ( 16 – 18%) fermentable material A ton should yield about 25 gallons of alcohol Material should be crushed to pulp & cooked for 2- 3 hrs, dilution is not necessary ( 79 – 80% water)
  • 13. PH is adjusted & fermentation is carried out Potato:- 15 -18% fermentable sugar A ton yield 20-25% of alcohol Cooked with steam under pressure After cooking mash is cooled to conversion for 15-20 min Once converted PH should be adjusted mash is fermented Sweet potato:- 27-28% fermentable sugar A ton should yield 40 gallons of alcohol Similar manner to potatoes Dilution is required (66%water)
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  • 15. Processing steps Starchy materials are of 2 categories 1. Encased by grain hulls 2. Starch is readily available Malting:- Seeds are moistened & allowed to sprout These seeds are powdered & the powder is called malt Milling:- Grains are milled to expose the starch Material is ground as fine as possible
  • 16. But the fine flour is difficult to remove in distillation process . Pot stills may reduce the problem Cooking (preparation of mash):- Starch is liquefied by boiling under pressure To dissolve the water soluble starches Always done with steam under pressure at 100°c Use less water during cooking & water may be added to dilute the mash So cooling time may be saved New method :- Milling & cooking in one operation without use of water  The cooked malt is called mash
  • 17. Conversion :- Conversion of starch to fermentable sugar (Maltose) by certain enzymes in the malt or by acid hydrolysis Enzyme extracts:- Commercially available 3 basic types : 1. α amylase - produce dextrose 2. Β amylase - produce maltose 3. Gluco amylase – reduce the remaining starch
  • 18. Acid hydrolysis:- Conversion by the action of acid Requires acid proof equipment and high temp Dilute mineral acid(H₂SO₄) is added to grain slurry Then immediately neutralized with calcium hydroxide Mash cooling:- Mash is cooled after conversion & PH is checked Cooling coils are used for cooling Cooling time is reduced as much as possible
  • 19. Fermentation Batch fermentation is commonly employed. Fermenter volume -- 600mᶟ. 3% inoculum(3×10⁶/ml) With in 12 days yeast produce 10% ethanol  when the process carried out at 35- 38°c , ph.4-4.5 max productivity was 1.9 g/h. 80%cells were removed in separator and brought back again into fermenter. When the high quality molasses is used, the max yield is 95%.
  • 20. Continuous fermentation not commonly used In US 10 large scale plants among them 4 are used con fermentation Nutrient solution was molasses with di ammonium phosphate PH was adjusted to 5 with H₂SO₄ Fermentation temp -35°c After fermentation cells were separated by centrifugation and channeled back into the first fermenter
  • 21.
  • 22. PRODUCT RECOVERY Cell biomass is separated by centrifugation or sedimentation. M.o desired end product – heated ( aggregate) readily separated from broth by sedimentation. Desired end product metabolite. intracellular - n.a , vit ,enz, a.b (gresofulvin) extracellular - a.a, acitic acid, citric acid, alcohol, [ penicillin, streptomycin]
  • 23. Separation of cell biomass Flocculation flotation Filtration centrifugation flocculation:- Flocculating agent is added hydrocolloids inorganic salts organic poly electrolyte  produce large aggregate which still settle more readily
  • 24. Flotation :- by introducing gas into liquid Cells gets adsorbed to the gas bubbles & rise to the foam layer at the top of vessel That can be collected & removed from the bioreactor Filtration:- Bacterial cultures – absolute filters  filamentous Fungi- depth filters static & cross flow filtration
  • 25. Distillation Separation for a mixture of liquids It relies on differences in B.P of component liquids to be separated The mixture to be separated is added to the distilled pot & is heated Low B.P will vaporize first. this vapors passes into distilling head & then into condenser. With in the condenser the vapor is cooled & it liquefies. The resulting liquid is then collected in receiving flask
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  • 27.
  • 28. Ethanol B.P - 78.5°C Water B.P - 100°C Purification:- By chromatography HPLC GLC Applications solvent in the manufacture of varnishes and perfumes Preservative for biological specimens In the preparation of essence and flavorings
  • 29. In many medicines and drugs. As a disinfectant and in tinctures. Used as a biofuel (99.2%). Medically ethanol is soporific. Used as antiseptic. Fluid in many alcohol thermometer. 92.4%ethanol is used as solvent in cosmetics pharmaceuticals and chemical industry Byproduct DDGS (distillers dried grain with solubles) Co₂– used for the production of dry ice and preparation of soft drinks Yeast biomass- animal fodder (contains proteins &vit)