Nei Pereira Jr., PhDFull Professornei@eq.ufrj.br

LADEBIO-EQ/UFRJWebsite: www.ladebio.org.br

LIGNOCELLULOSIC BIOMASS FOR THE PRODUCTION BIOETHANOL

WITHIN THE CONCEPT OF BIOREFINERY

FEDERAL UNIVERSITY OF RIO DE JANEIROSCHOOL OF CHEMISTRY

LABORATORY OF BIOPROCESS DEVELOPMENT

The Brazilian Context

BIOMASS is an attractive candidate as analternative to petroleum, since:

The only abundant source of renewable carbon;

Lower CO2 footprint than petroleum;

Domestically available.

Renewable % Non-renewable %

Hydropower 14.4 petroleum 39,1

Sugar cane 13.5 Natural gas 8.9

Wood and vegetal coal 13.2 Mineral coal 6.7

Others 2.7 Nuclear 1.5

Total 43.9.

56.1

Fonte: BEN/MME

Source Share of Primary Energy in Brazil (2005)

THE COUNTRY HAS ONE OF THE CLEANEST ENERGY MATRIX IN THE WORLD

What are Biomasses and Why to use them in substitution and/or in association to fossil sources ?

Natural Biomass

Food Biomass

Energetic Plantation Biomass

Residual Biomass

Plant Biomass

WATER

STARCH

CELLULOSE

SUGAR

CO2LIGHT

O2

ORGANIC MATTER from PLANT or ANIMAL sources;

Direct or indirect origin from the PHOTOSYNTHESIS PROCESS;

They are RENEWABLE.

Plant Biomasses

SUGARY LIGNOCELLULOSICSTARCHY

STARCH CELLULOSE HEMICELLULOSESUCROSE

XYLOSE MANNOSE

ARABINOSE

FRUCTOSE GLUCOSE

CARBOHYDRATES=

SUGARS=

SACCHARIDESGALACTOSE

Ranking dos Países Produtores de Etanol

0

20

40

60

80

2003 2004 2005 2006 2007 2008

Eta

no

l (m

ilh

õe

s d

e m

3)

Brasil USA China EU Índia

Fonte: LMC International (2008).

22,5 bilhões de litros

34 bilhões de litros

SUGAR STARCH LIGNOCELLULOSE

PRETREATMENTJuice Extraction

PRETREATMENTStarch solubilization

FERMENTATION• BATCH

• FED BATCH• CONTINUOUS

PRODUCT SEPARATION/ PURIFICATION

• DISTILLATION• SOLVENT EXTRATION• ADSORPTION

CELLULOSEHYDROLYSIS

STARCHHYDROLYSIS

sucroseglucosefructose

glucose

RESIDUEAnimal feeding

RESIDUEAnimal feeding

Energy production

xylosearabinosegalactoseglucose

glucose

LIGNINEnergy production

co2

ETHANOL

VINASSE

TECHNOLOGIES FOR ETHANOL PRODUCTIONFROM BIOMASSES

MATURE TECHNOLOGIES

EMERGING TECHNOLOGY

PRETREATMENT:EXTRACTION/

HEMICELLULOSEHYDROLYSIS

1st

Generation

2nd

Generation

TECHNOLOGICAL COMPLEXITY

1st GENERATION ETHANOLSUGAR CANE &

GRAINS/CEREALS

Fuels vs Food

LIGNOCELLULOSIC BIOMASS

3rd GENERATION ETHANOL ALGAE BIOMASS

Does not compete with food production

Faster growth than traditional crops;

Does not compete with agricultural cultures.

ETHANOL AND ITS DIFFERENT GENERATIONS

2nd GENERATION ETHANOL

The oil barrel prices reached valueswhich make unfeasible the self-sustainable growth of the nations;

Increasing interest for alternativesources of energy;

Necessity in aggregating value toresidual biomass, whose generationtends to grow;

Possibility for increasing the ethanolproduction without the need to expandthe availability of agricultural area forfeedstock production.

Importance of Bioetanol and its Production from Lignocelulosic Biomass

RESIDUAL BIOMASS OF

LIGNOCELLULOSIC COMPOSITION

Benefits of Lignocellulose Conversion Technologies through Biological and/or Chemical Routes for the Producition of

Fuels & Chemicals

Abundant and cheap renewable resources;

Their generation does not compete with the use of land for foodproduction;

No necessity of expanding the agricultural land for feedstockproduction;

Opportunity for industrial development based on the concept ofBIOREFINERY;

Reduction in gas emissions that cause the so called “green houseeffect”;

Cleaner technologies;

Macroeconomic benefits for rural communities and for Society as awhole;

They are within the perspective of Sustainable Development.

The Lignocellulosic Complex

Basic Composition :

Cellulose (40-60%)

Hemicellulose (20-40%)

Lignin (10-25%)

LIGNIN

HEMICELLULOSECELLULOSE

Main Differences between Cellulose and Hemicelluloses

CELLULOSE HEMICELLULOSES

Homopolysaccharide composed of GLUCOSE units

Heteropolysaccharides composed of several units of PENTOSES and HEXOSES

High degree of polymerization (2,000 to 18,000)

Low degree of polymerization (50 to 300)

Produces fibrous arrangements Do not produce fibrous arrangements

Presents crystalline and amorphous regions

Present only amorphous regions

Slowly hydrolyzed by diluted inorganic acid in high temperatures

Rapidly attacked by diluted inorganic acid in high temperatures

Is alkaline insoluble Are alkaline soluble

Corn cobs

Wheat straw

Rice straw

Sugar cane

bagasse

Cotton seed

Newspaper Urban residues

Carbohydrate (%)

Glucose 39.0 36.6 41.0 38.1 20.0 64.4 40.0

Mannose 0.3 0.8 1.8 n.d. 4.1 16.6 8.0

Galactose 0.8 4.4 0.4 1.1 0.1 n.d. n.d.

Xylose 14.8 19.2 14.8 24.3 4.6 4.6 14.0

Arabinose 4.2 4.4 4.5 4.5 4.3 0.5 4.0

Non-carbohydrate (%)

Lignin 15.1 14.5 9.9 18.4 17.6 21.0 20.0

Ashes 4.3 9.6 4.4 4.8 14.8 0.4 1.0

Proteins 4.0 4.0 n.d. 4.0 4.0 n.d. n.d.

Source: Lee (1997)

Composition of Some Lignocellulosic Residues

~ 70% carbohydrate

LIGNOCELLULOSIC BIOMASS

BIOCHEMICAL PLATFORM

• HYDROLYSIS (Chemical/Enzymatic)• LIGNIN CONVERSION

THERMOCHEMICAL PLATFORM

• PYROLYSIS (Bio-oil)• GASIFICATION BTL

GAS and LIQUIDintermediates

SUGARSintermediates and lignin derivatives

What is a Biorefinery?

BIOREFINERIES are similar to petroleum refineries in concept; however,biorefineries use biological matter (as opposed to petroleum or other fossilsources) to produce TRANSPORTATION FUELS, CHEMICALS, and HEAT andPOWER.

INDUSTRIAL BIOREFINERIES have been identified as the most promisingroute to the creation of a new industry based on renewable resources.

BIOFUELS(Ethanol; DMF; Gren Diesel; Gasoline and Kerosene)

CHEMICALS

POWER

BIOMASS

GLUCOSE

ETHANOL

ETHERS

ESTERS

REGENERATED

CELLULOSE

BUTANOL

ACETONE

GLYCEROL

CITRIC ACID

BUTIRIC ACID

GLUTAMIC ACID

LACTIC ACID

GLUCONIC ACID

SUCCINIC ACIDSCP

CELLULOSE

SORBITOL

HMF

ETHYLENE

BUTADIENEESTERS

SYNTETIC

RUBBER

(BIO)POLYMERS

VITAMIN C

FURANIC

POLYMERS LEVULINIC ACID

POLYESTERS

LC BIOREFINERY: Products from Celulose

POLY-HYDROXY-ALCANOATES

BIOPOLYMERS

ENZYMES

LCF: Lignocelulose Feedstock

DMFACETIC

ACID

Alcohol-chemistry

HEMICELLULOSE

BIOMASS

CELLULOSE

MONOSSACHARIDES (SUGARS)

XYLOSE; GLUCOSE; ARABINOSE; GALACTOSE; MANNOSE

SCP ETHANOL BUTANOL ACETONE FURFURAL

FURAN LYSINA

TETRA HYDRO FURAN

NYLON

ACETIC ACID URONIC ACID

LC BIOREFINERY:Products from Hemicellulose

ORGANIC ACIDS

PHA’s

GLUTAMIC

ACID

RESINS and

PLASTICS

XYLITOL

SORBITOL

MANITOL

GALACTOL

ARABITOL

ENZYMES

LCF: Lignocelulose Feedstock

DMF

HEMICELLULOSE

BIOMASS

CELLULOSE

SYNGAS

METHANOL

PHENOL,

VANILLIN,

OXIDIZED LIGNINBIO-OILS

PYROLYSIS

GASIFICATION

OXIDIZINGPROCESSES

PHENOLIC

RESINS

CHEMICAL

PRODUCTS

LIQUID

FUELS

SOLID FUEL

LC BIOREFINERY: Products from Lignin

LCF: Lignocelulose Feedstock

LIGNIN

ENERGY

liquid hydrocarbons similar to

those of petroleum crude oil

APPLICANT TOTAL COST DOE SHARE ANNUAL PRODUCTION

CAPACITY

PROJECT LOCATION

FEEDSTOCK Technology

VERENIUM 91,347,330 76,000,000

(granted 2007)

1,500,000 Jennings, LA Bagasse, energy crops, agricultural

wastes, wood residues

Biochemical

FLAMBEAU LLC 84,000,000 30,000,000 6,000,000 Park Falls, WI Forest residue BTL

ICM 86,030,900 30,000,000 1,500,000 St. Joseph, MO Switchgrass, forage

sorghum, stover

Biochemical

LIGNOL INNOVATIONS

88,015,481 30,000,000 2,500,000 Commerce City, CO

Woody biomass, agricultural

residue

Biochemical Organosolve

PACIFIC ETHANOL

73,040,000 24,340,000 2,700,000 Boardman, OR Wheat straw, stover, poplar

residuals

Biogasol(ETOH, biogas,

solid fuels)

NEW PAGE 83,653,212 30,000,000 5,500,000 Wisconsin, WI Wood biomass –mill residues

BTL

RSE PULP 90,000,000 30,000,000 2,200,000 Old Town, Maine

Wood chips (mixed

hardwood)

Biochemical

ECOFIN, LLC 77,000,000 30,000,000 1,300,000 Washington Country, KY

Corn cobs Biochemical (Solid State

Fermentation)

MASCOMA 135,000,000 25,000,000 2,000,000 Monroe, TN Swichgrass and hardwoods

Biochemical

TOTAL 808,086,923 305,340,000 25.2 M gallons = 95.4 M liters

Recently-awarded Projects (Small Biorefineries)

LIQUID BIOFUELS FROM LIGNOCELLULOSIC BIOMASS

POTENTIAL CANDIDATES

BIOFUELS Process Technological

maturity

BIOETHANOL Biochemical Emerging

BIOBUTANOL Biochemical Embrionary

DMF Hybrid Embrionary

FOSSIL LIKE GREEN FUELS Themochemical Growing

SOLUBLE LIGNIN

LIGNOCELLULOSIC

FEEDSTOCK

PRETREATMENT

(PRE-HYDROLYSIS/AUTO-HYDROLYSIS)

HEMICELLULOSE

(PENTOSES + HEXOSES)

CELLULOSE + LIGNIN

(CELLULIGNIN)

OH-

(DELIGNIFICATION)

CELLULOSELIGNIN

H+

(HYDROLYSIS)

GLUCOSE

FRACTIONING LIGNOCELLULOSIC BIOMASS FOR BIOCHEMICAL PLATFORM

Enzymatic Hydrolysis

TENDENCY

Inhibitors of Biological Processes GLUCOSE

Pretreatment of Lignocellulosic Feedstock

Aims at disorganizing the lignocellulosic complex, resulting in an increase of CELLULOSE DIGESTABILITY

(enzymes accessibility to cellulose molecules)

Lignin

Cellulose

Hemicellulose

Pretreatment

MAIN PRETREATMENT TECHNOLOGIES

(for solubilization and hydrolysis of HEMICELLULOSE)

STEAM EXPLOSION (needs further hydrolysis)

Sudden Decompression(to flash cool the biomass)

XYLAN CONTAININGLIGNOCELULOSIC

MATERIALS

High Steam Pressure

Liquid extracted by explosion, composed of:• Xylose ~ 10%• Xylooligosaccharides (DP 2 - 10) ~ 90%

• Uronic and acetic acids

Humid material with the lignocellulosiccomplex disorganized and with itsdigestibility increased

Simplified Scheme of the Steam Explosion Process

AMMONIA FIBER EXPANSION (needs separation of HEM from Lignin)

CATALYZED STEAM EXPLOSION (pretreatment associated to hydrolysis)

DILUTED ACID HYDROLYSIS IN MILD CONDITIONS (more used)

THERMOHYDROLYSIS (needs further hydrolysis)

Characteristics of the Main Pretreatment Technologies

for LC Feedstsocks (Biochemical Platform)

Characteristics Pretreatment Technology

SE CSE DAH TH (LHW) AFEX

Typical operational conditions

Batch or Continuous2 to 10 min

Batch or Continuous2 to 10 min

Batch or Continuous5 to 30 min

Batch5 to 60 min

Batch5-15% amônia

10-30 min

Temperature 190-270oC 160-200oC 150-180oC 170-230oC 100-180oC

Consumption of chemical inputs

No Yes Yes No Yes

Removal of hemicelullose

Yes Yes Yes Yes No, only after washing

Xylose yield 10% Xylose; 90% Xylosaccharides

70-90% 85-95% 50% Hemicellulose derived sugars

60% HDS after washing and removal

of lignin

Removal of Lignin Minor effect Moderate effect Moderate effect Minor effect Major effect

Formation of inhibitors

Yes, under severe conditions

Yes, under severe conditions

Yes, under severe conditions

Few Yes, under severe conditions

Reduction of the required particle size

Medium Medium High Medium High

Efficiency of the Cellulose Enz. Hydrol.

80-90% 80-85 % 80-85 % 80-90% 50-90 %

Waste Generation Less significant Moderate Significant Less significant Significant

Corrosivity of the medium

Low Low to moderate Moderate to high Low Low to moderate

Simplicity of the process (potential)

High Moderate to high Moderate Not evaluated Moderate

State of arte Pilot plants Pilot plants Pilot and demo plants

Bench scale Pilot plant

SE: Steam-explosion; CSE: Catalysed Steam-explosion; DAH: Diluted Acid hydrolysis; TH: Thermohydrolysis (liquid hot water pretreatment);AFEX: Ammonia fiber explosion. Sources: Adapted from LYND (1996); OGIER et al. (1999) and Balat et al. (2008); Pérez et. Al. (2007)

p-hydroxibenzoic acidm-hydroxibenzoic acid vanilinic acidsiringic acid p-hydroxibenzaldehydevanilincinamic acidsiringaldehydeconiferyl alcoholsinapyl alcohol

LIGNIN

Parajó et al. (1998)

hydroxy-methyl furfural furfural

HEMICELLULOSECELLULOSE

Substances that are commonly reported as inhibitors of the metabolic activity, originated from the acid

hydrolysis/pretreatment of lignocelulosic materials

acetic acidacetaldehyde

CHOHOH2C CHO

AMORPHOUSREGION

CRYSTALLINEREGION

1: ENDOGLUCANASE

CELLODEXTRINS

1

Enzymatic Hydrolysis of Cellulose

CELLULASES ARE INHIBITED BY CELLOBIOSE E GLUCOSE

CELLOBIOSE

2

2: EXOGLUCANASE 3: -GLUCOSIDASE

Cellulose

GLUCOSE

3

Why hydrolyse cellulose

enzymatically?

Milder conditions of pressure, temperature and pH;

High specificity;

Elimination of hydroxymethyl furfural, amongst other toxic substances (lignin derivatives);

Low energy consumption;

Low material costs with construction of equipments, differently of those processes which utilize acid hydrolysis.

R & D & I High production costs;

However,

Strategies for Ethanol Productionfrom Lignocelulosics

Following the Biochemical Platform

PRETREATMENT (HEMICELLULOSE

HYDROLYSIS)

ENZYMATIC HYDROLYSIS

OF CELLULOSE

CELLULASEPRODUCTION

C6FERMENTATION

DISTILLATION

C5FERMENTATION

ETHANOL

SOLUBLE SUGARS

solids (cellulignin)

SEPARATE HYDROLYSIS AND FERMENTATION

SHF

However, cellulases are inhibited by their final hydrolysis products

(CELOBIOSE and GLUCOSE)

PRETREATMENT (HEMICELLULOSE

HYDROLYSIS)

ENZYMATIC HYDROLYSIS

OF CELLULOSE

CELLULASEPRODUCTION

C6FERMENTATION

DISTILLATION

ETHANOL

SOLUBLE SUGARS

solids (cellulignin)

SIMULTANEOUS SACCHARIFICATION AND FERMENTATION

SSFENZYMATIC HYDROLYSIS

OF CELLULOSE &C6 FERMENTATION

Two Stream Fermentation

Model

C5FERMENTATION

PRETREATMENT (HEMICELLULOSE

HYDROLYSIS)

HIDRÓLISEENZIMÁTICADE CELULOSE

CELLULASEPRODUCTION

FERMENTAÇÃOC6

DISTILLATION

ETHANOL

solids (cellulignin)

SIMULTANEOUS SACCHARIFICATION AND COFERMENTATION

ENZYMATIC HYDROLYSISOF CELULOSE

&C6 FERMENTATION &C5 FERMENTATIONSOLUBLE SUGARS

SSCF

MolecularBiologyIntegrated

FermentationModel

PREATRETMENT(HEMICELLULOSE

HYDROLYSIS)

DISTILLATION

ETHANOL

CONSOLIDATED BIOPROCESSING

SOLUBLE SUGARS

CELLULASE PRODUCTION&

CELLULOSE HYDROLYSIS&

C6 FERMENTATION&

C5 FERMENTATION

CBP

solids (cellulignin)

Molecular Biology Consolidated

Bioprocess Model

Empresa País de origem Características de processo Localização Capacidade (m3/ano)

AE Biofuels EUA Hidrólise enzimática Montana 567

Blue Fire Ethanol EUA/Japão Hidrólise com ácido concentrado

CalifórniaIzumi

12.110n.d.

Chempolis Ou Finlândia Hidrólise com ácido diluído Oulu n.d.

Iogen Canadá Hidrólise enzimática Ontario 4.000

KL Energy EUA Hidrólise enzimática Wyoming 5.680

Lignol Energy Canadá Pré-tratamento Organosolv Vancouver 2.500

Mascoma EUA n.d. Nova Iorque 1.890

Poet EUA n.d. Dakota do Sul 75

Sekab Suécia Hidrólise enzimática n.d. n.d.

ST1 Finlândia n.d. LappeenrantaHaminaNärpiö

1.0001.0001.000

St. PetersburgState Forest-Technical Academy

Rússia Hidrólise c/ácido diluído 13 unidades nopaís

n.d

Sun Opta Canadá Hidrólise enzimática China n.d.

Universidade da Flórida

EUA Hidrólise enzimática com E. coli recombinantes (modelo integrado)

Flórida 7.570

Verenium EUA Hidrólise enzimática (modelo de duas correntes)

LouisianaJapão

5.3004.920

Empresas que empregam tecnologias para a produção de etanol de segunda geração (escala demonstrativa)

e características de processo

P-2

Utilidade Fria

Utilidade Quente

Água

Utilidade fria

Utilidade quente

CaO

BAGAÇO

VINHOTO

ETANOLCaSO

4

Pretr

eatm

en

t

HEMICELLULOSE HYDROLYSATE

SSF (Saccharomyces cerevisiae)

Alcohol Fermentation Pichia stipitis

Cellulase Production

CELLULIGNIN

PETROBRAS PILOT PLANT

UFRJ

Patent PI0505299-8 (11/2005)Patent PI0605017-4 (12/2006) (PATENT nb. 1000)

26/10/2007

Patent PI0200801-58 (12/2008)

2nd Generation Ethanol Pilot Plant

CENPES-PETROBRAS

2006 – 2009

Building of the pilot plant;

1st in Latin America;

More than 80 runs with bagasse;

Process reproductibity (bench scale pilot plant);

New projects in partnership to implement this technology in an industrial level.

Estágios

Desenvolvimento do tema “biorrefinarias: rota bioquímica”

no mundo

2010 - 2015 2016 - 2025 2026 - 2030

Comercialização

Produção/

processo

Inovação/

implantação

Scale-up

Fase demonstração

Fase piloto

Pesquisa em bancada

Estágios

Desenvolvimento do tema “biorrefinarias: rota bioquímica”

no mundo

2010 - 2015 2016 - 2025 2026 - 2030

Comercialização

Produção/

processo

Inovação/

implantação

Scale-up

Fase demonstração

Fase piloto

Pesquisa em bancada

T1c

T1cT1d

T1cT1d

T1dT1c

T1c

T1c

T1c

T1b

T1a T1d

T1eT1b

T1a

T1d

T1b

T1a T1d

T1e

T1a

T1a

T1a

Estágios

Desenvolvimento do tema “biorrefinarias: rota bioquímica”

no Brasil

2010 - 2015 2016 - 2025 2026 - 2030

Comercialização

Produção/

processo

Inovação/

implantação

Scale-up

Fase demonstração

Fase piloto

Pesquisa em bancada

Estágios

Desenvolvimento do tema “biorrefinarias: rota bioquímica”

no Brasil

2010 - 2015 2016 - 2025 2026 - 2030

Comercialização

Produção/

processo

Inovação/

implantação

Scale-up

Fase demonstração

Fase piloto

Pesquisa em bancada

T1e

T1e

T1d

T1d

T1d

T1b T1c

T1c

T1d T1c

T1b

T1d

T1c

T1b

T1d

T1c

T1bT1d

T1cT1a

T1a

T1a

T1b

T1b

T1a

T1b

T1cT1a

T1a

T1a

Mapa tecnológico do tema “Biorrefinarias: Rota Bioquímica” no Mundo e no Brasil

(2010 – 2030)

Notação: T1a – Pré-tratamento; T1b – Produção de celulases; T1c – Biologia molecular; T1d –Produção de biocombustíveis de segunda geração e de outras moléculas; T1e – Produção de energia e Integração energética de processo.

Challenges for efficient ethanol production from lignocellulosic biomass

Cost-effective pretreatment technology, with minimumgeneration of toxic substances (fermentation inhibitors);

Dedicated (in plant) cellulase production;

Enzyme engineering for efficient biomass hydrolysis;

Genetically modified microorganisms and processoptimization for efficient fermentation of C5 and C6;

Process energy integration (valorization of lignin).

Thanks to my “Army”, and Thanks for you attention !

Laboratories of Bioprocess Development

Federal University of Rio de Janeiro