APPLICATION OF NON-CONVENTIONAL YEASTS IN BIOPROCESSES

Sustainability is one of the most pressing challenge of our century, this term is becoming a main keyword of political agendas and more in general of mass media. To increase the “greenness of bioprocesses”, academia and industry, especially in the biotechnological and chemical fields, are focusing their studies with the scope to shift from traditional organic synthesis to new processes with reduced ecological foot-print. A good way to increase sustainability could be set up bioprocesses exploiting microorganisms. Nowadays, companies are searching new organisms that, differently from the well characterized Saccharomyces cerevisiae, show to be more resistant to the harsh conditions commonly occurring in industrial fermentations (high salt concentration, temperature and pressure). Due to their peculiar features, non-conventional yeasts (NCYs) seem to be a promising solution. On the other hand, the disadvantage to use these new organisms is related to the few studies and literature data available, especially compared to S. cervisiae. To fill this gap researchers have started to characterize these new species. My PhD work had dual aim: • First to identify good candidates, with specific physiological properties, that could be exploited in bioprocesses. • Second to characterize new promising enzymatic activities useful for industrial applications. In the first studies, I focused my attention on marine yeasts. I chose yeasts isolated from this environment, because their use gives the possibility to perform a seawater-based bioprocess saving large amount of fresh waters, reducing both cost and environmental impact. From our laboratory yeasts collection, I selected, for their halotolerance, two different Debaryomyces hansenii strains. Hence mechanisms involved in osmotic stress response have been investigated employing flow cytometry. I showed that hyper-osmotic stress elicits membrane depolarization and decreases membrane permeability to cationic compounds. This phenomenon reduces ions permeability and can negatively affect the uptake of charged substrate during bioprocesses. My research proceeded with the set up of new fermentation protocols in seawater-based media composed by a mixture of hexose and pentose sugar and cheap nitrogen sources. In these conditions we obtained high biomass yield (0.627) in 40 h of bioprocess. In the second part of my PhD project, I studied NCYs as sources of enzymes. With this aim I identified a nitrilase of marine strain of Meyerozyma guilliermondii, that displayed high activities on aromatic substrate, but also on arylaliphatic and aliphatic ones. These activities were maintained also in presence of high salts concentration. In particular M. guilliermondii nitrilase was able to perform complete dynamic resolution of mandelonitrile in seawaters within in 8 h. In the last part of my PhD, I identified a novel extracellular and cell-bound phytase activity in Cyberlindnera jadinii. This enzyme is suitable as feed additive, indeed activities at pH 4.5 and 37°C (animals gastric pH and temperature) were 26.25 mU/mgd.w. and 58.36 mU/mgd.w., detected as extracellular and cell-bound respectively. Phytase activities had their optimum at 50°C, reaching 37.2 mU/mgd.w. (extracellular) and 146 mU/mgd.w. (cell-bound). Data reported in my PhD work suggest that could be interest to proceed with further characterization on NCYs. New “green” bioprocesses characterized by high productivity could be a key for reach sustainability reducing the ecological impact of industrial production.