Agro-industry vegetable residues valorisation :
a waste biorefinery
Paola Di Donato
Associate Researcher
(paola.didonato@icb.cnr.it)
Institute of Biomolecular Chemistry
Via Campi Flegrei 34, (80078) Napoli - ITALY
November 20th, 2012 Città della Scienza, Naples, Italy
 Padova
31 institutes
Institute of Biomolecular Chemistry
Pozzuoli, Napoli
45 institutes
Rome 
Sassari 
Naples 
31 institutes
 Catania
November 20th, 2012 Città della Scienza, Naples, Italy
•Born in 1967 as “Istituto per la Chimica e Fisica delle Molecole di
Interesse Biologico” (ICMIB), “Institute for the Chemistry and Physics of
Biologically Interesting Molecules”.
• In 2002 it moved to the new building in Pozzuoli (NA) and took the
name of “Istituto di Chimica Biomolecolare (ICB-CNR)”. Institute of
Biomolecular Chemistry that today includes also the Roma, Catania,
Sassari and Padova branches.
• Main research activities in ICB are in the fields of organic chemistry,
biochemistry and microbiology with main topics including
environmental, pharmaceutical, food and biotechnological applications.
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry residues: a waste biomass
According the Directive 2001/77/EC of the European Community
biomass is “the biodegradable fraction of products, waste and
residues from agriculture (including vegetal and animal
substances), forestry and related industries, as well as the
biodegradable fraction of industrial and municipal waste”
Source: http://www.ecomena.org/biomass-resources-in-middle-east/
November 20th, 2012 Città della Scienza, Naples, Italy
Residual biomass: yearly production in Italy
BIOMASS
QUANTITY
Animal residues
150.000.000 t/y
Agro-industrial wastes
12.000.000 t/y
Slaughter wastes
1.000.000 t/y
Sewage sludges
3.000.000 t/y
Municipal wastes (organic fraction)
9.000.000 t/y
Forestry and crop residues
10.000.000 t/y
(source: C.R.P.A., Animal Production Research Centre, Italy)
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry residual biomass in Italy: yearly amounts
Biomass available
(thousand tons/year)
Actual re-use strategy
Wood transformation
4.400
Energy production
Wood recycling
8.000
Energy production
Paper Industry
270
Energy production
Food Oil
450
Energy, composting
Winery
450
Energy, composting
Canning and market packaging
340
Energy, composting
Rice transformation
330
Energy, composting
Industry
(source: Itabia )
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry residual biomass: a resource that can be valorised
Environmental problem
Economical issue
…. but a significant fraction of biomolecules is lost after
industrial processing, being discarded into the residual matter…..
proteins, fibres, polysaccharides, sugars, polyphenols, carotenoids
drugs
nutraceuticals
cosmetics
biopolymers
biofuels
 by-products that can represent the starting material for other processes
 raw materials not in competition with food chain
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry residual biomass: a resource that can be valorised
The biorefinery approach
• Food industry wastes
BIOMASS
Transformation
technologies
Energy and
chemicals
(vegetables processing residues)
• Agriculture and crop residues
•Bioprocesses (bacteria, enzymes)
•Chemical Processes
•Physical Processes
•Biofuels
•Antioxidants
•Biopolymers
Adapted from Kamm, B.; Kamm, M. (2004). Principles of Biorefineries. Appl. Microbiol. Biotechnol. (AMB), 64: 137-145.
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry wastes biorefinery: ongoing research
at the Institute of Biomolecular Chemistry
Crop production
Industry
Waste fraction
Tomato
(Lycopersicon esculentum)
5.125.000 t/y
Canning for juice and paste
1,8-2,3 %
(peels, seeds, immature and damaged fruits)
Lemon
(Citrus limon)
488.000 t/y
Liquor production
60 %
(peels, pulps)
Carrot
(Daucus carota)
630.000 t/y
Packing for whole sale market;
juices production
30-40 %
(immature or damaged fruits)
Fennel
(Foeniculum vulgare var.
Dulce)
600.000 t/y
Packing for whole sale market
40 %
(leaves, damaged fruits)
dry waste gross composition
Tomato
Lemon
Carrot
Fennel
Data from INRAN (Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione)
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry wastes biorefinery: ongoing research
at the Institute of Biomolecular Chemistry
Polysaccharides
recovery
II generation
bioethanol
Antioxidants
recovery
November 20th, 2012 Città della Scienza, Naples, Italy
Isolation of tomato’s wastes polysaccharides
Tomato waste peels
Chemical and physical
characterization of isolated
polysaccharides
Alkali treatment at r.t.
MW (Da)>1.000.000; High viscosity;
High thermal resistance; [a]D25-0,189.0
Polysaccharide
Glc/Xil/Gal/GalN/GlcN/Fuc (1:0.9:0.5:0.4:0.2:tr)
•Collaboration with Dr. Mario Malinconico and Dr. Barbara Immirzi (ICTP – CNR)
Container and object construction
Innovative material for containers or
for green house applications
November 20th, 2012 Città della Scienza, Naples, Italy
Isolation of tomato’s wastes polysaccharides
 biotechnological applications
Tomato waste peels
Alkali treatment at r.t.
Biodegradable film for solarization or
mulching applications
•Collaboration with Dr. Mario Malinconico
and Dr. Barbara Immirzi (ICTP – CNR)
Polysaccharide
 biological properties
Inhibition of NF-kB/DNA binding activity
Incorrect regulation of NF-κB has been linked to cancer, inflammatory and autoimmune diseases, et cetera
November 20th, 2012 Città della Scienza, Naples, Italy
vegetable wastes polysaccharides extraction: ongoing research
fennel residues
lemon wastes
(pomace)
alkali/acid extraction
•Structural analysis (MW; monomer
composition; spectroscopic analysis)
•Thermogravimetrical analysis
carrot residues
•Maceration
•Ultrasound assisted
•Microwaves assisted
POLYSACCHARIDES  Biotechnological applications
 Biological properties
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry wastes biorefinery: ongoing research
at the Institute of Biomolecular Chemistry
Polysaccharides
recovery
II generation
bioethanol
Antioxidants
recovery
November 20th, 2012 Città della Scienza, Naples, Italy
vegetable wastes polyphenols extraction: ongoing research
fennel residues
lemon wastes
(pomace)
MeOH; EtOH;
MeOH/H2O; EtOH/H2O
•Structural analysis
•Antioxidant and radical
scavenging activities
•Biological activity assay
carrot residues
tomato residues
•Maceration
•Ultrasound assisted
•Microwaves assisted
POLYPHENOLS
 Biotechnological applications
 Biological properties
November 20th, 2012 Città della Scienza, Naples, Italy
Vegetable wastes polyphenols extraction
work in progress…..
Project PON01_01966
Integrated agro-industrial chains with high energy efficiency for the development of
eco-compatible processes of energy and biochemicals production from renewable
sources and for the land valorisation (EnerbioChem)
Naples University
“Federico II”
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry wastes biorefinery: ongoing research
at the Institute of Biomolecular Chemistry
Polysaccharides
recovery
II generation
bioethanol
Antioxidants
recovery
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry wastes: production of enzymes for second
generation bioethanol from extremophiles
Bilateral Project Italy-Turkey
“Extremophiles for Next-Generation Biofuels ”
Dr. Barbara Nicolaus
Prof. Ebru Toksoy Oner
•Vegetable wastes under investigation: potato peels, carrot, tomato, sugar beet
pulp, starch and sugar beet molasses
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry wastes: production of second generation bioethanol
I generation bioethanol
Starch
(energy crops)
T = 90100°C
II generation bioethanol
Waste Biomass*
liquefaction by thermostable
amylases (starch cooking)
Simultaneous Saccharification
and Fermentation
dextrins
T=
65°C
saccharification by
thermostable amylases
glucose
Starch degradation and sugar
Fermentation by engineered yeasts:
expression of raw starch degrading
enzymes in Saccharomyces spp
glucose
T=
45°C
Yeasts Fermentation
(Saccharomyces spp)
CH3CH2OH
CH3CH2OH
* Not in competition with food chain
November 20th, 2012 Città della Scienza, Naples, Italy
Agro-industry wastes: production of enzymes for second
generation bioethanol from extremophiles
Amylolytic thermophile
Agro-industry wastes as culture media for
extremophiles’ fermentation
Geobacillus thermoleovorans
(strain PIZZOT)
dry cell weight (mg)
500
synthetic media
lemon
400
tomato
300
200
100
(starch digestion to glucose
without cooking pre-treatment)
Optical microscopy observation of raw starch before (a)
and after (b) digestion by G. thermoleovorans amylase
tomato
fennel
F
F1
B
D
F
D
F
F
F1
D
B
D
F1
F
D
B
F
D
F
F1
B
lemon
D
F
D
M
+G
a
D
M l
+T
D re
M
+M
al
raw starch degrading amylase
D
C
M
0
carrot
wastes as culture media for amylase production
Complex medium
Lemon medium
16.25 U/mg protein
6.375 U/mg protein
Di Donato P., G. Fiorentino G., Anzelmo G., Tommonaro G.,
Nicolaus B., Poli A. 2011 “Waste Biomass Valorization” 2: 103-111
November 20th, 2012 Città della Scienza, Naples, Italy
In conclusion…..
by means of an integrated biorefinery approach waste biomass
from agro-industry can be valorised to give
 Fibers and polysaccharides for biodegradable polymers
production
Biomolecules for several applications: antioxidants and
bioactive molecules for pharmaceutical, cosmetic or food
applications
Microorganisms and enzymes for second generation biofuels
production
November 20th, 2012 Città della Scienza, Naples, Italy
Thanks to….
Dott. Barbara Nicolaus
Dott. Annarita Poli
Dott. Licia Lama
Dott. Giuseppina Tommonaro
Prof. Ebru Toksoy Oner
“If you want to go fast, go alone. If you want to go far, go together”
African proverb
November 20th, 2012 Città della Scienza, Naples, Italy
Thanks for your
attention!
Dott. Paola Di Donato
paola.didonato@icb.cnr.it
paola.didonato@uniparthenope.it
Grönlund-Nisunen - Wave of Matter 2012