In the present work, brewers’ spent grain (BSG), which represents the major by-product of the brewing industry, was recovered from a regional brewery and fractionated in order to obtain a complete valorization. In particular, the whole process was divided in two main parts. A first pretreatment with hot water in an autoclave allowed the separation of a solution containing the soluble proteins and sugars, which accounted for 25% of the total starting biomass. This first step allowed the preparation of a medium that was successfully employed as a valuable growing medium for different microbial fermentations, leading to valuable fungal biomass as well as triglycerides with a high content of linear or branched fatty acids, depending on the microorganism used. The solid water-insoluble residue was then submitted to a lignocellulose deep eutectic solvent-mediated fractionation, which allowed the recovery of two important main fractions: BSG cellulose and BSG lignin. The latter product was tested as potential precursor for the development of cement water reducers with encouraging results. This combination of treatments of the waste biomass appeared to be a promising sustainable strategy for the development of the full exploitation of BSG from a circular economy perspective.

Recently, we described the preparation of the recombinant oleate hydratase from Lactobacillus rhamnosus ATCC 53103. We observed that the purified C-terminal His-tagged enzyme was completely inactive and the catalytic activity was partially restored only in presence of a large amount of flavin adenine dinucleotide (FAD). In the present work, we assess that this hydratase in the presence of the reduced form of flavin adenine dinucleotide (FADH₂) is at least one hundred times as active as in the presence of the same concentration of FAD. By means of two different biochemical processes, we demonstrated unambiguously that oleate hydratase from Lactobacillus rhamnosus ATCC 53103 is a FADH₂-dependent enzyme. As a first relevant application of this discovery, we devised a preparative procedure for the stereoselective synthesis of (R)-10-hydroxystearic acid. Accordingly, the hydration of oleic acid (up to 50 g/L) is performed on a multigram scale using the recombinant hydratase and FADH₂ generated in situ as cofactor. The produced (R)-10-hydroxystearic acid (ee > 97%) precipitates from the reaction solvent (water/glycerol/ethanol) and is conveniently recovered by simple filtration (>90% yield).

The use of Reactive Deep Eutectic Solvents (RDESs) in the preparation of polar head modified phospholipids (PLs) with phospholipase D (PLD)-catalyzed biotransformations has been investigated. Natural phosphatidylcholine (PC) has been submitted to PLD-catalyzed transphosphatidylations using a new reaction medium composed by a mixture of RDES/buffer. Instead of exploiting deep eutectic solvents conventionally, just as the reaction media, these solvents have been designed here in order to contribute actively to the synthetic processes by participating as reagents. RDESs were prepared using choline chloride or trimethyl glycine as hydrogen-bond acceptors and glycerol or ethylene glycol, as hydrogen-bond donors as well as nucleophiles for choline substitution. Specifically designed RDES/buffer reaction media allowed the obtainment of PLs with optimized yields in the perspective of a sustainable process implementation.

Different microbial strains are able to transform oleic acid (OA) into 10-hydroxystearic acid (10-HSA) by means of the catalytic activity of the enzymes oleate hydratase (EC 4.2.1.53). Lactobacillus rhamnosus ATCC 53103 performs this biotransformation with very high stereoselectivity, affording enantiopure (R)-10-HSA. In this work, we cloned, in Escherichia coli, the oleate hydratase present in the above-mentioned probiotic strain. Our study demonstrated that the obtained recombinant hydratase retains the catalytic properties of the Lactobacillus strain but that its activity was greatly affected by the expression procedure. According to our findings, we devised a reliable procedure for the hydration of oleic acid using a recombinant E. coli whole-cell catalyst. We established that the optimal reaction conditions were pH 6.6 at 28 °C in phosphate buffer, using glycerol and ethanol as co-solvents. According to our experimental protocol, the biocatalyst does not show significant substrate inhibition as the hydration reaction can be performed at high oleic acid concentration (up to 50 g/L).

The acid degumming waste of the seeds oil refining industry is currently disposed of, but, instead, it could be exploited as an important source for the preparation of many products. In this work, the waste coming from the soybean oil refining step was first recovered and treated, allowing the isolation of a fraction enriched of phospholipids (PLs). Then, the latter was transformed, via an enzymatic reaction catalysed by phospholipase D (from S treptomyces netropsis ), into more valuable products: polar head modified PLs‐enriched mixtures containing phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidylethanolamine (PE) and phosphatidylhydroxybutyrate (PB). In the following we show how biocatalysis can be exploited for the production of value‐added PLs, to be used as functional food and nutraceutical ingredients, from a renewable feedstock. This alternative to the industrial usual disposal procedures should impart to the whole process a greater value in terms of carbon recycling, in agreement with the paradigms of bioeconomy for a wiser reuse of renewable resources in a circular economy perspective.

The biotransformation of the aromatic amino acids phenylalanine, tyrosine and tryptophan originates a number of bioactive compounds. Yeasts are the most used microorganisms for the transformation of (L)-phenylalanine into the flavour phenylethanol. Here, we reported a study on the biotransformation of the proteogenic aminoacids phenylalanine, tyrosine and tryptophan by yeast strains belonging to Yarrowia genus. We found that the latter microorganisms, in high aerobic conditions, metabolise the aromatic amino acids (L)-phenylalanine and (L)-tyrosine with the almost exclusive formation of phenylacetic acid and 4-hydroxyphenylacetic acid, respectively. Differently, the biotransformation of (L)-tryptophan with Y. lipolytica, gave anthranilic acid as the main product. As stated by the European and USA legislations concerning natural flavour production, phenylacetic acid obtained by microbial conversion of phenylalanine of natural origin can be commercialised as a natural flavour. Accordingly, our findings were exploited in a new process, based on the Yarrowia strains-mediated biotransformation of natural (L)-phenylalanine, that allows the large-scale preparation of the high-value, natural flavour, phenylacetic acid.