UNIVERSITA’ DEGLI STUDI DI MILANO:
The Department of agricultural and environmental science of the University of Milan develops its activies in view of a development, through scientific research, of the knowledge regarding agricultural, forestry,
farming, environmental and energetic systems, based on a multi-disciplinal approach, dedicated to the
promotion of efficient management of complex agricultural systems. The ‘’Agriculture and Environment’’
and ‘’Biomass and Agro-energy’’ laboratories, are equipped with analytical sophisticated instruments like
Solid State NMR, DRIFT, GCMS, GC, GC MS-MS, HPLC, nanoscale porosimeter, elemental analyzer and
ICP/MS. The ‘’Biomass and Agro-energy’’ laboratory is equipped with lab-scale reactors for bioprocess
simulation and for microalgae growth. The research group counts also on a genetic and molecular biology
section. The research team is formed by 1 full professor, 3 researchers and 2 post-docs. They have
developed in the last 25 years, knowledge in the field of biomass use and re-use, with particular attention
to energy production. More in detail, the research group has developed, in joint with private firms,
innovation in the field of biogas production. Recently, the first full-scale plant for bio-hydrogen and biomethane integrated production has been developed, unique in Italy and EU. Besides, another full-scale
application has been developed, based on chemical-physical separations, for renewable nutrients
production from waste biomass. The research team results integrated with different activities about the
production of 2nd and 3rd generation biomass (Arundo Donax and microalgae), activities on full-scale
plants for bio-energy production, analytical activities, genetic characterization and applications of
molecular techniques and engineering. The contribution of UNIMI research group is fundamental. First of all the project relies on the availability of 2nd and 3rd generation biomass, which will be produced by the
UNIMI group.

Project DANCE, funded by Cariplo Foundation
Microalgae have taken recently importance as they are able to produce high added value products (antioxidant, proteins and poly-unsatured fatty acids) to be used in different productive sectors, i.e. food, feed, pharmaceutical, nutraceutical and cosmetic industries. Microalgae production till now has been developed under autotrophic condition that shows limitation in terms of biomass (and products) yield and economic feasibility. On the other hand, many interesting microalgae are able to growth under heterotrophic condition by using organic C sources. Again, this condition does not allow to produce light induced high added value molecules. In this way, mixotrophic condition, i.e. autotrophy plus heterotrophy represents the good solution allowing both high biomass and high value molecules production. The market of high valuable molecules from microalgae is continuously increasing (worth between 3.5 and 5 billion €), as they represents the base for many products made by food, feed, pharmaceutical, cosmetic and nutraceutical industries.
Agro-food system in Italy is one of the most developed in EU and it has been estimate that about 200 million cubic meters per year of polluted wastewaters are produced (EUROSTAT, 2014). Agro-industrial waste streams need to be disposed safety and economically, and they represent and additional cost for industries. On the other hand, because of their origin waste streams from agroindustry are interesting because they represent safe renewable C-sources and safe renewable nutrients (e.g. N, P and K plus meso- and microelements), that could be usefully recovered to develop a third generation biorefinery, i.e. no- food biomasses (waste) from a productive process becomes a feedstock for a successive productive process.
The proposed project (DANCE) has the aim to recover renewable C and nutrient sources from Lombardy agro-industry to be used to produce microalgal biomass rich in high added-value molecules by developing mixotrophic bio-technology. Added-value products will be isolated from algae biomass by using green technologies, i.e. supercritical CO2 extraction, allowing safe residual to be recovered to produce plant bio-stimulants and bioenergy, developing a biorefinery concept. All project goals well match with Cariplo foundation requirements.
Chosen algae have been selected according to their ability to growth under mixotrophic condition. Basing on literature data Chlorella sp., Nannochloropsis sp., Phaeodactylum tricornutum and Haematococcus pluvialis had been chosen for their ability to produce strongly marketable compounds, i.e. proteins, essential aminoacids, total lipids, fatty acids with particular emphasis on ω3, ω6, and carotenoids (astaxanthin above all). In order to perform mixotrophic condition, different waste streams from agri-food activities will be considered. Dairy, winery and animal farming waste streams, will be used as they are largely and widely available in Lombardy Region. In addition glycerol, a C-rich renewable-waste coming from biodiesel production, will be used when waste requires available-C addition (e.g. animal farming). Wastes will be characterized before their use and adjusted, in order to meet the algae trophic needs. Waste streams pre-treatment will be performed, also, when required, in order to get microalgae needs in terms of C and nutrients availability, e.g. polymers and/or oligomers hydrolysis and or fermentation to get “ready to be used” monomers. After chemical composition optimisation, algae will be grown in batch conditions under mixotrophic and autotrophic conditions. Microalgae will be cultivated in 500 mL cylindrical photobioreactors and illuminated with fluorescent day-light lamps (6400 K) with a mean irradiance of 80 µmol photons m-2 s-1. Air bubbling will provide the necessary culture mixing. Optimal microalgae pH will be controlled via chemical buffers.
Algae growing parameters and molecules production will be measured using UV spectrophotometer (optical density) and HPLC-GC/MS approaches.
The most promising algae vs stream will be up scaled to semi-continuous cultivation conditions to simulate industrial production for each molecule class. Sustainable extracting technology (supercritical CO2) will be applied. After molecules extraction, residual algal biomass will be considered as feedstock to produce bio-stimulating complexes for plant, because of the presence of high amount of hormones and hormone like molecules that have been showed by scientific literature to stimulate plant growth. To do this different concentration of algal biomasses will be tested on germination phase of hormone-less mutant plant. When hormone-like activity will be found a chemical characterisation of the biomass will be performed using HPLC and GC/MS. On the other hand when no hormone like activity will be demonstrated, residual biomass will be used to produce bioenergy (anaerobic digestion) giving a correct hierarchy of products value (from high value to low value products) coming from the EU biorefiney concept.
Environmental impacts and C footprint of the entire biorefinery will be measured by Life Cycle Assessment (LCA) comparing 1) state of the art of waste streams disposal, 2) autotrophic algae cultivation conditions, 3) mixotrophic algae cultivation conditions, focusing on the positive effect of recovering renewable resources by biorefinery. Results of DANCE will be publically available to stakeholders and public administration through an online data suite and dissemination activities.

Country: Italy

Organization Type: University

Email: barbara.scaglia@unimi.it

City: Milano ,Via Celoria, 2 Google map