An overview of the main
research activities of the
“Biomass and Bioenergy Lab”
Speaker: Cristiano Varrone
Email: [email protected]
Giulio Izzo, Giulia Massini, Antonella Signorini, Fabrizio De Poli, Antonella Marone,
Floriana Fiocchetti, Andrea Aliboni, Chiara Patriarca, Silvia Rosa, Luciano Mentuccia,
Elena De Luca, Cristiano Varrone.
Laboratory of Biomass and Bioenergy
Head of Laboratory: Dott. Giulio Izzo / [email protected]
IDROBIO Project: coupling dark and light fermentation
- 2.4 L bioreactor operated at 80 °C and inoculated withThermotoga neapolitana ; a yield of 3.9 mol
H2/mol glucose was obtained, with a production rate of 51 mL/L/h
- 3.8 L bioreactor with 1.8 L working volume (BFM medium; 10g/L glucose, 37°C; pH 6.8),
inoculated with 1g of sediments from the Averno lake; a yield of 1.7 mol H2/mol glucose was
obtained, with a production rate of 1.8 L H2/L/day and 54.6% of H2 content
50 L Tubular Photobioreactor tested
under natural light conditions (in
cooperation with UO CNR-ISE) , using
Rp. palustris 42 OL grown on a synthetic
medium . Mean H2 production rate = 17
mL/L/h.
50 L bioreactor for the culture
of T. neapolitana , with 25 L
working volume.
1.2 L planar Photobioreactor (in
cooperation with UO ENEA) with a surface
of 0.0730 m2 containing Rp. palustris
AV33 isolated from Averno sediments.
Max H2 production rate : 1,4 L / L / day
Continuous Flow Reactor using a mixed pools
l H2/ l day mol H2/mol gluc. max rate (l H2/l h)
Continuum 1300 h
7-8
2-2,5
0.5- 0.6
H2 (%)
54-59
Dark Fermentation of waste products
Hydrogen/biogas production by dark anaerobic fermentation of organic wastes is a
promising strategy to obtain renewable and clean energy in a sustainable way
It can lead to the conversion of organic waste and feedstock
into a host of valuable chemicals and energy
The Approach:
One way to improve the efficiency of H2 production is to explore the potentials
offered by the microbial biodiversity, both in natural and artificial environment,
identify and to select bacterial strains with high H2 producing abilities from
different substrates, and to characterize the microbial metabolism, in order to
understand and optimize the whole process.
Synthetic Overview
HYDROGEN PRODUCTION FROM:
 Isolation of hydrogen producing bacteria from vegetable
waste for bioaugmentation: 2.1-2.4 L/L/d
 Selection and acclimatation of microbial mixed pools for
degradation of agri- and zootecnical waste: 2.8-3 L/L/d
 Bioconversion of crude glycerol into H2 and ethanol
from enriched activity sludge: 3 L/L/d
MSE-ENEA project: H2 production from vegetal waste
isolation and characterization of meso-philic bacterial strains
contained in the waste for bioaugmentation
Vegetable
waste
Isolation of bacterial strains
Selection of
H2-producers and
cellulolytic bacteria
1 – V (leaf shaped vegetable waste);
2 – VP (80% leaf shaped vegetable
waste + 20% potato peels);.
DNA
extraction
Test on cellulose and
hydrolysis products
glucose
cellobiose
Identification:
Sequencing of amplified
16S rDNA
arabinose
xylose
PCR amplification
16S rDNA gene
Improvement of the H2 production from self-fermentation of
Vegetal Waste performing Bioaugmentation
90
V
70
60
50
40
30
20
10
0
0
5
10
Buttiauxella sp.4
15
20
25
30
Fermentation time (h)
Rahnella sp. 10
Raoultella sp. 47
35
40
Consortium
45
50
Self-fermentation
90
VP
80
Cumulative H2 production (ml H2 /gVS)
Cumulative H2 production (ml H2 /gVS)
80
Effect of the artificial
consortium
on vegetal waste
70
60
50
40
30
20
10
0
0
5
Buttiauxella sp.4
10
15
20
25
30
Fermentation time (h)
Rahnella sp. 10
Raoultella sp. 47
35
40
Consortium
45
50
Self-fermentation
Marone et al., 2012 International
Journal of Hydrogen; 37(7): 56125622.
Substrati
Inoculo
H2 (%)
L H2/l/d
ml H2/g VS
m3 H2/t substrate
Vegetal waste
Self-fermentation
10-12
0.557-0.748
18-22
1-1.3
Vegetal waste
Bio-agumentation
26-28
2.1-2.4
67-86
4-4.8
 Bioconversion of agri- and zootechnical waste
Degradation of Organic Matter :
1°Phase
Hydrolysis of
macromolecules
2°Phase
Digestion of
hydrolisates
3°Phase
4°Phase
Acidogenesis Methanogenesis
Vegetables
Wood
BIOMASS
H2/CO2
Sugars
Aminoacids
Fatty acids
Manure Glycerol
•
•
•
•
Theoretic Yield from Glucose
C6H12O6 → 3CH 4+ 3CO2
C6H12O6 + 2 H2O → 2CH3COOH + 2CO2 + 4H2
2CH3COOH → 2CH 4 + 2CO2
C6H12O6 + 2 H2O → 2CO2 + 2CH 4 + 4H2
BIOGAS
CH4/CO2
Carboxylic
acids
Alcohols
Acetate
Energetic efficiency
83.2%
33.5%
89.0%
Marea project: coupling H2 and CH4 production
from agri- and zootecnical waste
Substrati
Cheese whey
Cow manure
Glycerol
Inoculo
consorzio
IDROBIO
consorzio
IDROBIO
consorzio
IDROBIO
H2 (%)
44-46
L H2/l/d
2.8-3.0
10-12 0.220-0.230
ml H2/g
VS
m3 H2/t
substrate
160-180
20-23
23-25
0.520-0.540
32-35 0.600-0.800 120-140
Al bruciatore
Al bruciatore
Idrogeno,CO2
Liquam i
Glicerolo
Serb. Alim.
Alim entazione
Enhanced production of methane by bioaugmentation
of H2 producing community
Bioreatt ore 2
Q
Bioreatt ore 1
Pompa 2
Acqua Risc./Raff
Pompa 1
Q
3.86 L/Ld
Brodo in
ferm entazione
Q
Mixture Design
(H2 = 34-40%)
Microorganism i
m etanogeni
Microorganism i
idrolitici
30-33
C.M = 66%,
C.W. = 33%
2.15-2.98 L/Ld
Metano, CO2
Al B ioreattore 2
Pompa 3
Digestato
Statistical Optimization of Glycerol Fermentation
In optimized conditions it was possible to obtain a max hydrogen production rate of more than
2.150 L H2/L/day (yield > 0.94 mol H2 / mol crude glycerol), while reaching a max EtOH
concentration of almost 8g/L (yield ~ 1), without adding any vitamins, minerals, triptone or
yeast extract.
H2 concentration in the biogas reached more than 50%, and the H2/medium ratio (mL/mL)
was found to be around 4 (max value obtained was 4,5 at 18g/L of glycerol).
Cristiano Varrone et al.,2012. International Journal of Hydrogen Energy
(in press, http://dx.doi.org/10.1016/j.ijhydene.2012.02.106)
Scale-up Tests
3 L BioFlo 115
Benchtop Fermentor
Pmax
Rmax
l
(mL)
(mL/h)
(h)
Novaol
3720.9
154.1
5.64
0.998
2997.7
0.91
Itabiol
3428.1
146.7
4.20
0.984
2983.4
0.89
Pure Gly
3808.1
138.8
6.34
0.998
2706.5
0.91
N.S. Novaol
3451.3
174.4
6.48
0.996
3152.4
0.92
R2
Rate
(mL/L/d)
Yield
(mol/mol)
Modified Gomperzt
equation
Rate=
Pmax/(l+Pmax/Rmax)
Future project: Pilot Plant
Alla "torcia"
We are now evaluating the possibility to set up
a 1000L pilot plant, which might produce up to:
Idrogeno
Acqua
Glicerolo
Microorganism i
Etanolo (95 %)
- 400-500Kg EtOH/t of glycerol
(with an estimated value of 350€/t glycerol)
Bioreattore
- 200-250 m3 H2/t of glycerol
(with an estimated value of 70€/t glycerol;
based on national subsidies of 0,28 cents/kWh)
Serb. Alim .
Pom pa 2
Acqua + ac. organici
Pom pa 1
Acqua+etanolo+acidi organici
Simulated industrial plant showed a production of 100 m3 H2/d and 25000 L EtOH/d,
correspoing to 80 GWh/anno from H2 and 540 GWh/anno from EtOH,
with an energy efficiency of 39%.
PATENT APPLICATION: Bioconversion of crude glycerol into hydrogen and ethanol
(Number: RM2011A000480)
Applying data: 13/09/2011. Inventor: Cristiano Varrone
Link: http://brevetti.enea.it/tabella.php (ENEA patent nr. 735)
ENTERPRISE EUROPE NETWORK:
http://portal.enterprise-europe-network.ec.europa.eu/ (Reference: 12 IT 56Z7 3PF3)
THANK YOU FOR YOUR ATTENTION