LIGNOCELLULOSICS-DERIVED CARBOHYDRATES …...Italian stakeholders meeting on biorefineries, Sassari,...

LIGNOCELLULOSICS-DERIVED CARBOHYDRATES AS

PLATFORM MOLECULES FOR THE PRODUCTION OF

BIOFUELS AND BIOBASED PRODUCTS

Isabella De Bari

ENEA CR TRISAIA, ITALYItalian Representative Task IEA 42

Intertasks Workshop andItalian stakeholders meeting on biorefineries, Sassari, 5 May 2015

� Test fields of energy crops� Thermochemical processes� Advanced biofuels� Biorefineries and green

chemistry

Combustion technologies

� Microalgae growth, � Anaerobic processes

to CH4 and H2

� Energy crops

ENEA’s research centres working on bioenergy/biorefineries

LCA – Anaerobic digestion

ITALIAN BIOMASS AVAILABILITY

agricultural area was reduced by 28% in 40 years from 18 million hectares to 13 (abandonment and cementification)

The forest area has increased in 40 years of 100% from 5.5 to 11 million hectares

A recent analysis is detailed in the Sector Plan for Bioenergy (5 August 2014)

by the Permanent Conference for relations between the State, the Regions and

the Autonomous Provinces

The estimated annual potential of

residual biomass amounts to about

30 million tons (dry matter) ≈10

Mtoe.

The agricultural areas for dedicated

crops are less than 200,000 hectares

but could be increased by 5 times

without conflicting with food

production.

Dedicated crops for biofuels and biobased products

Giant reed(Arundo donax)

Lignocellulosic.residues fromCardoon

Glucan 34.75 35

Xylan 20.10 14

Galactan 0.27 1,7

Arabinan 2.12 2,2

Mannan 0 1,1

Lignin Klason 22.0 25

Ash 7.70 8,1

EtOH Extractives 10.22 4,1

BIOLYFE "Second BIOethanol process:

demonstration scale for the step of

LignocellulosichYdrolysis and Fermentation

SEEDS

VEGETABLE

OILS

METHODS TO DISSOLVE LIGNOCELLULOSICS

SUGARS PLATFORM� breakdown of the raw materials into sugars � fermentation,

dehydration, hydrogenation

METHODS to dissolve lignocellulosic feedstocks: acid hydrolysis, , and

liquefaction in ionic liquids.

IMPROVING THE PERFORMANCES

THE MAIN STEPS IN THE BIOCHEMICAL ROUTE

12

3

BIOMASS

HYDROLYZATE

1. Optimizazion of the pretreatment2. Development of high gravity bioprocesses3. Biomass derived sugars�fermentation, dehydration, hydrogenation….6

PRETREATMENT

ENZYMATIC

HYDROLYSIS

ENEA’S TECHNOLOGIES FOR THE BIOMASS PRETREATMENT

ENEA’ CONTINUOS PLANT (300 kg/hr)

ENEA’ BATCH PLANT

Few % of additives �catalyses the process at milder conditions / affects the biomass

fractionationsBIOMASS+ ACIDS Hemicellulose mostly

hydrolized to monomers

I.e. Monomer sugars for fermentation, or dehydration

BIOMASS + BASES Hemicellulose mostly

olygomeric

I.e. olygomeric sugars to

produce barrier films

BIOMASS PRETREATMENT AND FRACTIONATION

WOODWaste paper

STRAW

ARUNDO

DONAX

GRAPE STALKS

HEMICELLULOSE

LIGNIN (residual

sugars ≈ 2%)

CELLULOSE

(≈80%)

PRETREATMENT FRACTIONATIONATION

Plant size 300 kg/h

Lignocellulosic biomass

Process severity

CELLULOSE HYDROLYSIS YIELD

(LOW SOLIDS LOADING)

time

Temperature

PRETREATMENT OF WOODY BIOMASS (ASPEN CHIPS)

Before

After

ACID-CATALYZED STEAM PRETREATMENT OF ARUNDO

DONAX [1,4%WT]

C5

REC

OV

ER

Y:

70

%

T[°C], t(min)

Klason

lignin ARABINAN Galactan Glucan Xylan

% COMPOSITION OF THE WATER-INSOLUBLE FRACTION

196°C+acid 27.1 0.69 0.21 36.0 8.6

200°C+acid 25.7 0.50 0.20 35.1 6.0

205°C+acid 23.5 0.52 0.18 34.9 4.8

210°C+acid 23.0 0.40 0.120 33.7 3.5

% OF SOLUBLE CARBOHYDRATES

196°C+acid 0.55 0.31 1.70 6.0

200°C+acid 0.80 0.40 2.4 9.5

205°C+acid 0.63 0.36 2.5 5.2

210°C+acid 0.51 0.25 1.7 4.3

Level of microbial inhibitors

4-HBAa

(mg/kg)

SYRb

(mg/kg)

5-HMFb

(mg/kg)

2-FURd

(mg/kg)

CATe

(g/kg)

F. acidf

(g/kg)

A. acidg

(g/kg)

196°C+acid 64 77 86 538 2.9 4 8.5

200°C+acid 112 77 128 747 4.9 7 15.6

205°C+acid 98 47 86 700 5.3 7.1 18

210°C+acid 194 126 35 954 1.4 8.4 23.9

a 4-hydroxybenzaldehyde. b Syringaldehyde. c 5-(Hydroxymethyl)furfural. d 2-Furaldehyde. e catechol. f formic acid. g acetic acid

The effect of these degradation products on the hydrolizates

fermentability will depend on the microorganism and process

set-up(inoculum size, pH, and process strategy)

HIGH GRAVITY HYDROLYSIS

• Effect of the pretreatment on the biomass hydrolizability

• High gravity hydrolysis of biomass • Hybrid simultaneous saccharification and

fermentation

High gravity hydrolysis = process in which the solids content is above 20%

High gravity hydrolyis

30

50

70

90

0 10 20 30

s/L [%]

xylo

se y

ield

, %

%

20

40

60

80

100

0 10 20 30s/L [%]

glu

cose

yie

ld [

%] 144 hr 48 hr

The Challenges of high gravity hydrolysis: �High viscosities � mass transfer limitations� poor mixing

�Inhibition by end-products

The Challenges of high gravity hydrolysis: �High viscosities � mass transfer limitations� poor mixing

�Inhibition by end-products

The Challenges of high gravity fermentation: � High concentration of microbial inhibitors� Osmotic stress due to high solutes concentration� Toxic effect of ethanol (synergistic inhibition)

EFFECT OF HIGH SOLIDS CONTENT ON THE BIOMASS HYDROLYSIS

Research activities

1.Investigate bioreactor geometries suitable for the processing of concentrated slurries

2.Optimize the bioreactor feeding strategies (bach , fed-batch)

3.Optimize the use of enzymatic mixtures (dosage, effect of auxiliary components, effect of surfactants)

4.Find the optimal process strategy (SSF, SHF, hybrid process)

5.Reduce the hydrolyzates toxicity to the fermentation microrganisms and to grow resistant microrganisms

EFFECT OF THE MIXING

1 2

3

Mixing in shaken flasks and bioreactor (1 and 2)

Arundo fiber, 20% solids loading, 50° C.

pH 4.8 pH 5.5

MIXING GEOMETRY

0

20

40

60

80

100

120

0 25 50 75 100

Time [h]

Glu

cose

, [g

/L]

GRAVIMETRIC

MIXING STIRRED TANK

Gravimetric shaking in rotating drum system was much more effective (Arundo fiber)

Mixing in stirred bioreactor and gravimetric shaker (2 and 3)

HYBRID HYDROLYSIS AND FERMENTATION

SHF: separate hydrolysis and fermentation

SSF: simultaneous saccharification and fermentation

19

SSFbiomass

H-SSF : hybrid simultaneous

saccharification and fermentation

� Optimization depends on the subsequent

fermentation strategy (i.e. fermentation or

cofermentation, type of microrganism).

� the liquefaction time is an important variable.

Production of bioethanol through hybrid process

Enzyme dosage

[g/g biomass

DM] MicrorganismsProcess

TypeYeast

inoculation T [°C]

Ethanol

(%wt)

0,07 S. cerevisiae SHF B 32°C 3,5

0,07 S. cerevisiae SSF B 32°C 3,9

0,027 S. cerevisiae HSSF B 37°C 3,7

0,027 S. cerevisiae HSSF FB3 50-37°C 3,8


0,07 K. marxianus H SSF FB 2 50-32°C 4,7

0,07 K. marxianus H SSF B 32°C 4,4


0,07 S. cerevisiae HSSF B 37°C 4,2

0,07 S. cerevisiae H SSF B 32°C 4,6

0,07 S. cerevisiae H SSF FB 2 50-32°C 5,0

B=batch; FB= Fed-Batch

Inhibitor Conc.

(g/L)

Yeasts Strain

Reduction of

ethanol yield (%)

Reduction of of ethanol

productivity (%)

5-HMF 4 S.cerevisiaeTembec T1

12 45

5-HMF 4 S.cerevisiaeCBS 8066

40

5-HMF 4 S.cerevisiaeY-1528

11 40

Furfural 4 S.cerevisiaeCBS 8066

69

Furfural 1.6 S.cerevisiaeTembec T1

27

Furfural 1.6 S.cerevisiaeY-1528

25

Acetic acid

4.3 S.cerevisiae 50

Aceticacid

8 P.stipitis 98

Microorganisms inhibition thresholds

Value

added

product

Microrganism Inhibition

threshold

Butanol Clostridium

beijerinckii

Xylitol Candica

Tropicalis

Lactic acid Lactobacillus

acidophilus

Biosuccinic

acid

Actinobacillus

succinogenes

Tryacyl-

glycerides

Cryptococcus

curvatus

………………

…..

………………………

…….

………………

…

…………………….

Microorganisms resistance to the biomass degradation products is an important requirement

for the conversion of lign. cellulosic derived carbohydrates to biobased products

PRODUCTION OF CLEAN SUGARS

� ION-EXCHANGE RESINS

� OVERLIMING

� STEAM STRIPPING

� ……………….

Hydrolyzates detoxyfication is sometime necessary to increase the microbial conversion efficiency

ADAPTATION

Fermentation

Detoxification =rimoval of degradation by-products (i.e. organic acids, furan

compounds, phenols)

Pretreated biomass/hydrolyzates

22

� MICROBIAL ADAPTATION

� FERMENTATION AT HIGH CELLS CONCENTRATION (IMMOBILIZATION)

Microbial adaptation

23

FERMENTATION WITH CO-IMMOBILIZED CELLS OF S. Cerevisiae and P. stipitis

Yeast(s) conditions

Process scheme P/S

% xylose consumption at

maximum ethanol

% glucose consumption at ethanol maximum Y%

COF 4 16 100 68COF 4 16 100 66COF 0,25 16 100 64COF 0,25 16 100 63COF 20 97 68COF 17 100 60COF 17 93 56COF 33 100 61COF 31 100 53COF 34 100 51COF 67 100 49COF 4 34 100 72COF 4 42 100 80COF 0,25 41 100 64COF 0,25 33 100 71SEQ 10^9 [S] 1,36x10^9 [P] 4 73 100 61

SEQ 10^9 [S] 5x10^9[P] 4 39 100 69

P. and S. adapted +

EtOH

Beads uptake

1,25,E+09P. adapted up to 40%

WILD4,70E+08

6,80E+08

P. adapted up to 40% (repeated

cycles)

3,25E+06

3,70E+07

4

COF= cofermentation

SEQ= sequential fermentation 24

EUROPEAN PROJECTS ON 2G BIOETHANOL

Project Main Partners

5FP BIO-H2: Production of clean hydrogen for fuel

cells by reformation of bioethanol

CR FIAT, ECN, Un. Patrasso,

Queens, CNRS, Peugeot, Renault

BIOPAL “Algae as raw material for production of

bioplastics and biocomposites contributing to

sustainable development of european coastal

regions”

CEVA, FIAT, Un. Pisa,

DEMOKRITOS, OWS

TIME “Technological Improvement for ethanol

production from lignocellulose”

VTT, Un. Lund, Roal, Un. Budapest,

Nedalco,

7FP BIOLYFE "Second BIOethanol process:

demonstration scale for the step of

LignocellulosichYdrolysis and Fermentation"

Biochemtex, ENEA, Novozymes,

Lund University, IFEU, WIP

Running (7FP) GRAIL “Glycerol Biorefinery Approach for the

Production of High Quality Products of Industrial

Value”

IUCT, ENEA, In.Bio-Consorzio,

STUBA, Megara, Biozoon, CENTIV,

DBFZ, Universidad Valparaíso, PI,

SINTEF, Queen’s University Belfast,

Aalborg University

Italian projects on biofuels/biobased

productsFunding

Ministry

Project

acronymFULL TITLE

MIPAAF MULTISORGO Integrated production of bioethanol and biogas

from sweet sorghum: technological, economic, energy and

environmental aspects

FITOPROBIO Phytodepuration treatments using cellulosic biomass to obtain

second generation ethanol"

BIOSEGEN Innovative chain for the production of second generation

biofuels from agricultural and agro-industrial

residues and biomass crops.

MSE

INDUSTRIA

2015

PRIT Development of an Italian pretreatment technology for the

production of second generation bioethanol (COORDINATED BY

BIOChemtex)

MIUR BIT3G

REBIOCHEM

ALBE

research activities within the green chemistry cluster -spring

ENEA, University of Salerno, Department of Industrial Engin eering(A. Giuliano, D. Barletta)

Process Flowsheet Analysis and Synthesis

for a Lignocellulosic Biorefineryproducing ethanol and biobased

products

Lignocellulosic Biorefinery Flowsheet: base case

BIOMASS PROCESS PROCESS ENERGY PRODUCTS

BIOMASS STEAM

EXPLOSION

ENZYMATIC

HYDROLYSISETHANOL

FERMENTATION

ETHANOL

DISTILLATION AND

PURIFICATION

LIGNIN

COMBUSTION

WASTE WATER

TREATMENT

HEMICELLULOSE

UPGRADING AND

FERMENTATION

LIGNIN

WASTE

WATER BIOGASHEMICELLULOSE

RICH

0,00

0,20

0,40

0,60

0,80

1,00

1,20

1,40

1,60

1 2 3 4 5

*PSPEnzyme in situ

/PSPBaseProcess

Enzyme price (€/kg)

Sensitivity on enzyme price

Xylitol is a platform molecule

Scenario with xylitol co-production


BIOMASS STEAM

EXPLOSION

ENZYMATIC

HYDROLYSISETHANOL

FERMENTATION

ETHANOL

DISTILLATION AND

PURIFICATION

LIGNIN

COMBUSTION

WASTE WATER

TREATMENT

LIGNIN

WASTE


RICH WASTE

WATER

HEMICELLULOSE

UPGRADING AND

FERMENTATION

PRELIMINARY ESTIMATIONS FROM OUR EVALUATIONS

Furans are platform molecules

Source: ( Angew 2007)

O

O

H

O

OCH3

O

O

OH

O

HO

O

Polymer

Biofuel

Fine chemical

Furfural

O

O

HHO

5-Hydroxymethylfurfurfual

O

O

H

Furfural

Scenario with furfural co-production


BIOMASS STEAM

EXPLOSION

ENZYMATIC

HYDROLYSISETHANOL

FERMENTATION

ETHANOL

DISTILLATION AND

PURIFICATION

LIGNIN

COMBUSTION

WASTE WATER

TREATMENT

LIGNIN

WASTE


RICH WASTE

WATER


COMPARISONC5 XYLITOL/FURFURAL

SCENARIO 1 2

Main Product Ethanol Ethanol

Co-Product Xylitol Furfural

Payback selling price for the co-product through our simulation(€/kg)

2 3

Estimated market price (€/kg)* 3.5 0.9-1.3

Tot Yield (tPRODUCTS/tBIOMASS ) 25 % 24 %

Electricity balance(MWPROD – MWCONS)

8.1 -4.7 (heat integration is

necessary)

Required Steam (t/h) 84 122

Succinic Acid is a platform molecule

HEMICELLULOSE

UPGRADING AND

FERMENTATION

Scenario with succinic acid co-production


BIOMASS STEAM

EXPLOSION

ENZYMATIC

HYDROLYSISETHANOL

FERMENTATION

ETHANOL

DISTILLATION AND

PURIFICATION

LIGNIN

COMBUSTION

WASTE WATER

TREATMENT

LIGNIN

WASTE


RICH

SUCCINIC ACID

FERMENTATION

SUCCINIC ACID

PURIFICATION

SUCCINIC ACID

CO2


WASTE

WATER

Scenario with succinic acid and xylitol co-production


BIOMASS STEAM

EXPLOSION

ENZYMATIC

HYDROLYSISETHANOL

FERMENTATION

ETHANOL

DISTILLATION AND

PURIFICATION

LIGNIN

COMBUSTION

WASTE WATER

TREATMENT

LIGNIN

WASTE


RICH

SUCCINIC ACID

FERMENTATION

SUCCINIC ACID

PURIFICATION

SUCCINIC ACID

CO2

PRELIMINARY EVATUATIONS FROM OUR ESTIMATIONS

Scenario with succinic acid/furfural co-production


BIOMASS STEAM

EXPLOSION

ENZYMATIC

HYDROLYSISETHANOL

FERMENTATION

ETHANOL

DISTILLATION AND

PURIFICATION

LIGNIN

COMBUSTION

WASTE WATER

TREATMENT

LIGNIN

WASTE


RICH

SUCCINIC ACID

FERMENTATION

SUCCINIC ACID

PURIFICATION

SUCCINIC ACID

CO2

WASTE

WATER


Scenario with ethanol+succinic acid co-production:COMPARISON C5 ETHANOL/XYLITOL/FURFURAL

SCENARIO 1 2 3

Main Product Ethanol Ethanol Ethanol

Co-Products Succinic Acid Succinic Acid + Xylitol

Succinic Acid + Furfural

Payback selling price for the co-product (RED) through our simulation (€/kg)

5.4 2.6 3.5

Estimated market price (€/kg)*

2.7 3.5 0.9-1.3

Tot Yield (tPRODUCTS/tBIOMASS ) 23 % 24 % 24 %

Electricity balance(MWPROD – MWCONS)

-4.1 -5.2 -5.5

Required Steam (t/h) 129 129 159

The biorefinery is no longer self sufficient for the energy need. A detailed LCA is necessary

40

Concluding remarks

CHALLENGES

Capitalization of the knowledge developed in the sector of biofuels for

the maximum exploitation of the “biomass barrel”.

� Optimize multi-products biorefineries by improving the existing

pioneering technologies

� Develop new processes for the conversion of side streams from

lignocellulosic biorefineries

� Improve the process integration

ENEA’s FACILITIES FOR BIOREFINING

41

� Pretreatment and fractionation at pilot scale

(300 kg/h)

� Production of second generation sugars

� Process scale up/downstream processing

� Technological platforms for thermal valorization

of biomass residues (pyro-gasification

� Identification of new proteins and key enzymes

involved in specific substrate degradation

/biomass degradation (proteomics) RECENT ACTIVITIES

� Fully equipped analytical labs for materials

characterization and process analysis

THANKS FOR YOUR ATTENTION

[email protected]

+39 0835974313

LIGNOCELLULOSICS-DERIVED CARBOHYDRATES …...Italian stakeholders meeting on biorefineries, Sassari,...

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Transcript of LIGNOCELLULOSICS-DERIVED CARBOHYDRATES …...Italian stakeholders meeting on biorefineries, Sassari,...