Progress on enzyme technology for bioethanol production
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Progress on Enzyme Technology for Bioethanol Production. International Symposium Delhi Gang Duan, PhD Surendra Bade & Jay K. Shetty, PhD Mar 20-21, 2008 . Agenda. Trends in bioethanol production New enzymes from Genencor Conclusion.

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Progress on enzyme technology for bioethanol production

Progress on Enzyme Technology for Bioethanol Production

International Symposium Delhi

Gang Duan, PhD

Surendra Bade

&

Jay K. Shetty, PhD

Mar 20-21, 2008


Agenda

Agenda

  • Trends in bioethanol production

  • New enzymes from Genencor

  • Conclusion


Ethanol and ddgs

Ethanol and DDGS

129 Plants are in operation

77 Plants are under construction


Progress on enzyme technology for bioethanol production

CORN TO ETHANOL

129 Plants are in operation and 77 Plants are under construction

  • Currently 6.0 billion gal of ethanol is produced in the US.

  • Estimates indicate that ethanol production in the US could be 12.0 billion gals/yr by 2009.

  • Most of the increase in the ethanol capacity will come from new dry grind ethanol plants

  • Supply of distillers dried grains with solubles (DDGS) will increase proportionately

1/3

+

1/3

+

1/3

*1lb=0.45kg


Progress on enzyme technology for bioethanol production

Starch To Ethanol

-Reaction Mechanism-

Enzyme+Energy

SUBSTRATE + H2O ---------------------------> PRODUCT

(Starch)Alpha Amylase (SPEZYME Fred,SPEZYME Xtra)(Dextrins)

Enzyme

SUBSTRATE + H2O ---------------------------> PRODUCT

(Dextrins)Glucoamylase (Optidex L400, Fermenzyme)(Glucose)

Yeast

SUBSTRATE ----------------------------> PRODUCT

(Glucose) Protease (FERMGEN) (EtOH,CO2,Biomass)


Conventional ethanol production process

pH adjustment steps are not shown

*

Conventional Ethanol Production Process

Thermo-Stable

Alpha Amylase

Alcohol

Recovery

Glucoamylase

Yeast

Milo

Corn

Wheat

Rye

Barley

Tapioca

Distillation &

Dehydration

Liquefaction

Saccharification

Fermentation

Water

JET COOKER

>100° C

5–8 MIN

*

STORAGE

TANK

60° C

8–10 HRS

(optional)

SLURRY

TANK

GRINDING

SECONDARY

LIQUEFACTION

95° C

~90 MIN

*

DDGS


Issues for the conventional processes

Issues for the conventional processes

  • EnergyEnergy

  • EfficiencyEfficiency

  • EnvironmentalEnvironmental

Enzyme

ENZYME SOLUTIONS


Corn to ethanol manufacturing cost 2005 dry milling plant we make enzymes work for you

Phytic acid removal and NSP modification

Reduced Energy Consumption

E

N

Z

Y

M

E

E

N

Z

Y

M

E

E

N

Z

Y

M

E

Improved Fermentation efficiency

Corn to Ethanol: Manufacturing Cost 2005 (Dry Milling Plant): We make enzymes work for you

Depreciation

Labor

Enzymes, yeast etc

Utilities

Net corn

DDGS

Ref: Gopal Chotani,2005 , Internal Communication ,Genencor-Danisco.


How do we create value using enzymes

How Do we Create value using enzymes ?

Process Development based on existing unit-operations

never results in radical process innovation

-Enzymes to reduce number of Unit operations (saving on the capital cost)

-Enzymes’ Kinetic energy replacing the thermal energy in the process ( reduction in energy cost-STARGEN Platform )

-Enzymes’ Kinetic energy replacing mechanical energy ( saving on capital cost and energy cost-Macerating and viscosity reducing Enzymes,Protease , STARGEN)

-Enzymes for enhancing de-watering effect-saving on drying

-Enzymes resulting in value added co-products (manufacturing cost reduction,For example Phytase for value added DDGS)


Factors that affect enzyme activity

FACTORS THAT AFFECT ENZYME ACTIVITY

  • Time

  • Temperature

  • pH

  • Substrate

  • Metal ions required for some enzymes

  • Inhibitors

  • Enzyme concentration


Factors affecting yeast performance

Factors affecting yeast performance

  • Microbial

    • Lactic acid (0.8% or 8g/L inhibitory. Overcome by higher inoculum)

    • Acetic acid

    • Ethanol

    • pH

  • Non microbial

    • Sulfite

    • Sodium

    • Temp.(Heat stress)

    • Nutrient stress


Progress on enzyme technology for bioethanol production

Anaerobic yeast process

Ethanol production

C6H12O6 2C2H6O + 2CO2 + 2ATP + Heat (32 kcal)

Cell growth

C6H12O6+ 1.2*0.63NH3 + xATP  0.63C6H10.8O3N1.2 + 2.22CO2 + 4 NAD(P)H

Glycerol production

C6H12O6+ 2NAD(P)H + 2ATP  2C3H8O3

  • From the above, if GA limits glucose concentration, then, it also limits ethanol production

  • Ethanol determines ATP production; cell growth and glycerol determine ATP consumption

  • However, anaerobic cell growth produces reducing power (NAD(P)H)

  • Nitrogen source: ammonia means more reducing power production

  • Nitrogen source: amino acids (AFP) mean less reducing power production

  • And, glycerol production recycles NAD(P)H, i.e., glycerol is proportional to cell growth

  • Hence net Ethanol yield on glucose is determined by growth and glycerol


Yeast growth alcohol production

Yeast Growth & Alcohol Production

12 enzymes involved synthesis of ethanol


Theoretical ethanol yield

THEORETICAL ETHANOL YIELD

  • Goal: Gallons of Ethanol per bushel of corn

  • Starch content in corn:

  • 56 Lbs Corn/Bu X 86% Ds (1) X 73% Starch (2) = 35.2 Lbs Starch/Bushel

  • Glucose content in starch

  • 35.2 Lbs Starch/Bu X 1.11 (3) = 39.1 Lbs Glucose/Bushel

  • Ethanol content based on glucose

  • 39.1 Lbs Glucose/Bu X .51 Lbs Ethanol/lb Glucose (4) = 19.9 Lbs Ethanol/Bu ÷ 6.54 (5) Lbs Ethanol/Gal = 3.1 Gal Ethanol/Bushel

  • (1) 14% Moisture In Corn

  • (2) Starch Content Of Corn

  • (3) Chemical Gain Of Starch To Glucose

  • (4) Lbs Ethanol Per Lb Glucose

  • (5) Density Of Ethanol

*1lb=0.45kg

1bushel=56 lbs=25.2kg


Theoretical yield corn as an example

THEORETICAL YIELD/Corn as an example

1 gal.

0.1 gal.

1 gal.

1 gal.

Bushel = 56 lbs. corn

1000kg corn at 14% moisture and 73% starch results in 455L Alcohal

Ethanol = 19.90 lbs. = 35.5%

CO2 = 19.20 lb. = 34.3%

DDGS = 16.9 lbs. = 30.2%

*1lb=0.45kg

1bushel=56 lbs=25.2kg


Actual ethanol yield

ACTUAL ETHANOL YIELD

  • Theoretical ETHANOL yield = 3.1 GAL/BU (455 L/MT)

  • Actual ETHANOL yield

    • Typically 2.6-2.8 Gal/Bu (90% +/- Efficiency) (~410 L/MT)

    • Factors reducing yield

      • Grain quality, moisture content, starch content

      • Starch conversion rate to glucose (bound)

      • Glucose conversion rate to ethanol (yeast)

      • Plant process control

*1lb=0.45kg

1bushel=56 lbs=25.2kg


Full product line for ethanol production

Full Product Line for ethanol production

  • Liquefying enzymes

    • SPEZYME XTRA: High performance, thermostable, fast viscosity-reducing

    • Maxaliq-one: A new liquefying enzyme and system with additional phytase

    • Spezyme Fred: Thermostable, low calcium, alpha-amylases.

  • Saccharifying enzymes

    • Distillase L-400, Optidex L-400, Optidex L-300;

    • Fermenzyme L-400, Glucoamylase with acid fungal protease;

  • Acid Fungal Proteases(Enhancing the yeast performance, shorten the fermentation time, increase the alcohol yield)

    • FERMGEN

    • GC 106

  • Viscosity reducing enzymes (Higher DS, easy to handle; potential more fermentable sugars)

    • OPTMASH BG, OPTIMASH TBG

    • OPTIMASH VR

  • Enzymes for no-cook process: STARGEN


Liquefying enzyme

Liquefying enzyme

Liquefaction

  • Solubilizes starch

  • Hydrolyzes starch to dextrins with enzymes

  • Reduces viscosity

    >SPEZYME XTRA

  • Thermostable

  • Fast viscosity reducing

    >MAXALIQ

  • Superior viscosity reducing

  • more DDGs values


Effect of phytase on viscosity reduction using maxaliq one

Effect of Phytase on Viscosity ReductionUsing Maxaliq One


Improving the value of ddgs by using phytase

Improving the value of DDGs by using phytase

Comparison of Conventional hot cook process and PALS (Phytase Amylase Liquefaction System) on the ethanol yield and phytic acid content


Saccharifying enzymes

Saccharifying enzymes

  • Saccharification enzymes benefit fermentation

    • Faster fermentation times

    • Higher ethanol yields

    • Improved yeast health

    • Better solids separation in the centrifuge

    • Reduces evaporator fouling

    • Less recycle or backset

    • Higher protein in DDGS

    • More consistent plant operations

      >DISTILLASE L-400

      >FERMENZYME L-400


Acid fungal protease

Acid Fungal Protease

  • FERMGEN


Viscosity reducing enzymes

Viscosity reducing enzymes

Viscosity reducing enzymes to hydrolyze the non-starch polysaccharides in wheat, sorghum and tuber before/during the cooking.

  • OPTIMASH BG for wheat/Barley

  • OPTIMASH VR for sorghum/Rye and others

  • OPTIMASH TBG, a thermostable beta-glucanase could be effective at 80o C.


Enzyme for no cook process

Enzyme for no-cook process


Stargen enzyme for no cook process

STARGEN ----Enzyme for No-cook process

Alcohol

Recovery

STARGEN

Yeast

Rice/Milo

+

water

Saccharification

Liquefaction

Water

JET COOKER

>120° C

5–8 MIN

Distillation &

Fermentation

*

STORAGE

TANK

SECONDARY

LIQUEFACTION

95° C

~90 MIN

60° C

3–10 HRS

(optional)

SLURRY

TANK

GRINDING

DDGS

No cook process using STARGEN:

-NO LIQUEFACTION; NO NEED for HEATING as well as for COOLING

-Very low sugar concentration in the system;

Relaxed and happy yeast by spooling;

Less contamination.

-More alcohol from raw material.

Overall, a much more simplified process, and more importantly, a more EFFICIENT process .


The difference stargen could make

The difference STARGEN could make:

Comparison of the amounts of alcohol produced from conventional and STARGEN process:

High temperature jet cooking:

Very good process - 400 liter

Good process - 390 liter

Average process - 380 liter

STARGEN process without any pretreatment

Produces 430-460 liter or even higher!


Benefits of stargen 001 in alcohol production in no cook process

Benefits of STARGEN 001 in alcohol production in No cook process

  • Increased Carbon Conversion Efficiency — Higher Yield

  • Energy Saving -Elimination of Jet Cooking

  • Reduction in Osmotic Stress/Reduction in By-products Formation — Glycerol, Organic Acids, etc.

  • Capacity Increase — High Density Fermentation - Higher Alcohol Yield

  • Reduction of Yeast Growth Inhibitors — High Glucose, Maillard Products, etc.

  • Saving on Operational Cost — Labor, Time, Chemicals

  • Elimination of Calcium Addition — Reduction of Calcium Salt Formation

  • Value Added By-product (DDGS) — Higher Protein Content

  • Process Simplification — Reductions in Unit Operations

  • Saving on Capital Cost — Capacity Increase/New Plant


Danisco genencor wuxi application centre delhi technical service lab

FINLAND

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Hanko

NETHERLANDS

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BELGIUM

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CHINA Beijing

Wuxi

JAPAN

Tokyo

USA

Rochester, NY

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SINGAPORE

ARGENTINA

Arroyito

Buenos Aires

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R&D

Admin.

Danisco / GenencorWuxi Application CentreDelhi Technical Service Lab

India


Acknowledgement

Acknowledgement

THANKS to all the team members in Wuxi Application Centre Lab for their hard and excellent work!

Sophia Xu, Kathy Qian, John Zhou, Jessica Li, Bruce Ruan

Raj Lad

Roy Sim


Progress on enzyme technology for bioethanol production

Danisco (India) Pvt.Ltd.

DLF Corporate Park

5thFloor,Block4B,Phase III,

Gurgaon-122 002

Attn.:--Mr.Yogesh Grover

Cell No. --+91 9999333505

Tel. No.–0124 4061510

Danisco (India) Pvt.Ltd.

302,Centre Point,

J.B.Nagar,NearKohinoor Continental Hotel,

Andheri-KurlaRoad,Andheri-E,

Mumbai-400 059

Attn.:--Mr.Sunil Kasat

Cell No.--+91 9833465865

Tel.No.–022 28258713

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