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The present agrifood production system can not sustainably feed the present and the future world population, and it needs to be re-designed in a circular economy perspective. Insects, with their ability to grow on agrifood by-products and wastes, yielding highly nutritious, and safe, food and feed ingredients, can help the transition to a more sustainable agrifood production. In the European Union, since 2018 insects have been designed as novel food, and their use as feed ingredients has been regulated. Insects are slowly, but steadily, becoming an integrated part of our agrifood system., BioLaw Journal - Rivista di BioDiritto, N. 2 (2020)

Because of the continuous increase in the prevalence of gluten-related disorders, selection of wheat with a low content of immunogenic gluten epitopes could be an innovative alternative for prevention. In this review, the focus is on literature data concerning the deallergenization tools of wheat, which are mainly related to breeding approaches (classic and advanced) and processing operations (germination and fermentation). Until now, no safe wheat genotype has been identified, whereas decreasing wheat allergenicity is possible. On the other hand, the decrease of gluten or some of its epitopes can strongly affect technological properties. Thus, obtaining celiac-safe gluten without affecting the technological properties of wheat could be considered as a new challenge that scientists will be facing. Celiac-safe wheat-based product development could be a great revolution in the market of foods for special medical purposes. The present paper is aiming to: (a) review the strategies and the approaches used, or that can be used, for developing low allergenic wheat: their utilities and limits were also discussed and (b) screen the impact of gluten reduction or removal on the quality of wheat end-use products.

Introduction: Phlorotannins are complex mixtures of phloroglucinol oligomers connected via C-C (fucols) or C-O-C (phlorethols) linkages. Their uniformity in subunits and large molecular weight hamper their structural analysis. Despite its commercial relevance for alginate extraction, phlorotannins in Laminaria digitata have not been studied. Objective: To obtain quantitative and structural information on phlorotannins in a methanolic extract from L. digitata. Methodology: The combined use of 13C and 1H NMR spectroscopy allowed characterisation of linkage types and extract purity. The purity determined was used to calibrate the responses obtained with the colorimetric 2,4-dimethoxybenzaldehyde (DMBA) and Folin-Ciocalteu (FC) assays. Using NP-flash chromatography, phlorotannin fractions separated on oligomer size were obtained and enabled structural and molecular weight characterisation using ESI-MS and MALDI-TOF-MS. Results: The fucol-to-phlorethol linkage ratio was 1:26 and the extract was 60.1% pure, determined by NMR spectroscopy. For DMBA, the response of the extract was 12 times lower than that of phloroglucinol, whereas there was no difference for FC. By accounting for differences in response, the colorimetric assays were applicable for quantification using phloroglucinol as a standard. The phlorotannin content was around 4.5% DM. Fucol- and phlorethol-linkage types were annotated based on characteristic MSn fragmentations. Structural isomers of phlorotannins up to a degree of polymerisation of 18 (DP18) were annotated and identification of several isomers hinted at branched phloroglucinol oligomers. With MALDI-TOF-MS phlorotannins up to DP27 were annotated. Conclusion: By combining several analytical techniques, phlorotannins in L. digitata were quantified and characterised with respect to fucol-to-phlorethol linkage ratio, molecular weight (distribution), and occurrence of structural isomers.

The possibility to obtain allergenic proteins by means of total chemical synthesis would be a big step forward in the development of cures to food allergy and in the study of the mechanism of allergic reactions, because this would allow to achieve control at the molecular level over the structure of the product and to study its relationship with the allergenic activity in fine details. This is instead not possible by using allergens produced by extraction from natural sources or by recombinant DNA techniques. In this work, we aimed to test for the first time the feasibility of the total chemical synthesis of an allergenic protein. Pru p 3, the most studied member of the family of lipid transfer proteins, relevant plant food pan-allergens, was used as model target. Strategies for the convergent assembly of the target protein, starting from five peptide fragments to be bound by means of either native chemical ligation or peptide hydrazide ligation, followed by desulfurization, to achieve ligations at alanine, were developed and tested. All the reaction conditions were set up and optimized. Two large peptides covering the two halves of the protein sequence were synthesized and structurally characterized by means of circular dichroism, and their immunogenicity was proved by means of immunoblot, using antibodies against Pru p 3, and immunoCAP inhibition tests. Finally, the five peptides were bound together to produce the whole protein stretch. The obtained results demonstrate the feasibility of total chemical synthesis as a new way to obtain pure allergens. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd

‘Management of waste as a resource’ was described in a European roadmap (COM (2011/571) as a milestone to be reached by 2020. On the one hand, there are diverse under-spent side-streams situated at the farmers and post-harvesting level as well as in the associated retail sector. On the other hand, there is a need for new resources, for instance proteins as alternative for soy proteins. InDIRECT, a three-year EU research project started November 2016, aims to develop biorefinery processes as part of new value chains to convert under-spent side-streams from the agro-sector and processing sector into useful marketable products. Cascading processes (recovery of multiple compounds from the same feedstock) are envisioned to increase the conversion efficiency and maximise the values of the feedstock. Direct bio-refiney of the biomass will be compared with indirect biorefinery. The latter refers to the two-step process that can cope with the heterogeneity of side-streams. In a first step the heterogenic feedstock is converted to homogenous biomass using insects. Insects are able to convert a variety of feedstocks into a more homogenous biomass, being their own biomass. In a second step, the insect biomass is further fractionated into a lipid, protein and chitin fraction, that all three have potential towards marketable end prod-ucts. The direct approach (without the insect conversion step) which is being explored for green leaves, is expected to be more seasonally variable compared to the indirect biorefinery appraoch, but generates a larger variety of compounds. The project foresees to process the derived fractions further into multiple marketable products for the feed, food and chemical sector. The multidisciplinary InDIRECT consortium consists of 9 partners from four countries (Belgium, The Netherlands, France & Italy), comprising 2 research organisations (VITO & University of Parma), 5 industrial partners (Nutrition sciences NV specialized in feed additives, two insect farms Millibeter & Proti-Farm R&D, IMPROVE specialized in plant proteins, and CHEMSTREAM focusing on chemical application), a non-profit organisa-tion that works in close contact with farmers and processing industry (Innovatiesteun-punt) and a project supporting company (Temperio). The project (www.bbi-indirect.eu) is coordinated by VITO. Session 1 Evaluation of the use of insects Bornimer Agrartechnische Berichte I Heft 97 I ISSN 0947-7314 24 This project has received funding from the Bio Based Industries Joint Undertaking under the European Union’s Horizon 2020 research and innovation programme under grant agreement No 720715.

‘Management of waste as a resource’ was described in a European roadmap (COM (2011/571) as a milestone to be reached by 2020. On the one hand, there are diverse under-spent side-streams situated at the farmers and post-harvesting level as well as in the associated retail sector. On the other hand, there is a need for new resources, for instance proteins as alternative for soy proteins. InDIRECT, a three-year EU research project started November 2016, aims to develop biorefinery processes as part of new value chains to convert under-spent side-streams from the agro-sector and processing sector into useful marketable products. Cascading processes (recovery of multiple compounds from the same feedstock) are envisioned to increase the conversion efficiency and maximise the values of the feedstock. Direct bio-refiney of the biomass will be compared with indirect biorefinery. The latter refers to the two-step process that can cope with the heterogeneity of side-streams. In a first step the heterogenic feedstock is converted to homogenous biomass using insects. Insects are able to convert a variety of feedstocks into a more homogenous biomass, being their own biomass. In a second step, the insect biomass is further fractionated into a lipid, protein and chitin fraction, that all three have potential towards marketable end prod-ucts. The direct approach (without the insect conversion step) which is being explored for green leaves, is expected to be more seasonally variable compared to the indirect biorefinery appraoch, but generates a larger variety of compounds. The project foresees to process the derived fractions further into multiple marketable products for the feed, food and chemical sector. The multidisciplinary InDIRECT consortium consists of 9 partners from four countries (Belgium, The Netherlands, France & Italy), comprising 2 research organisations (VITO & University of Parma), 5 industrial partners (Nutrition sciences NV specialized in feed additives, two insect farms Millibeter & Proti-Farm R&D, IMPROVE specialized in plant proteins, and CHEMSTREAM focusing on chemical application), a non-profit organisa-tion that works in close contact with farmers and processing industry (Innovatiesteun-punt) and a project supporting company (Temperio). The project (www.bbi-indirect.eu) is coordinated by VITO. Session 1 Evaluation of the use of insects Bornimer Agrartechnische Berichte I Heft 97 I ISSN 0947-7314 24 This project has received funding from the Bio Based Industries Joint Undertaking under the European Union’s Horizon 2020 research and innovation programme under grant agreement No 720715.

ISBN:978-3-907234-75-4

Cheese microbiota contribute significantly to the final characteristics of cheeses due to the growth and interaction between cheese microorganisms during processing and ripening. For raw milk cheeses, such as Parmigiano Reggiano (PR), the microbiota derive from the raw milk itself, the dairy environment, and the starter. The process of cheese making and time of ripening shape this complex ecosystem through the selection of different species and biotypes that will drive the quality of the final product by performing functions of their metabolism such as proteolysis. The diversity in the final peptide and amino acid composition of the cheese is thus mostly linked to the diversity of this microbiota. The purpose of this study was to get more insight into the factors affecting PR cheese diversity and, more specifically, to evaluate whether the composition of the bacterial community of cheeses along with the specific peptide composition are more affected by the ripening times or by the cheese making process. To this end, the microbiota and the peptide fractions of 69 cheese samples (from curd to cheese ripened 24 months) were analyzed during 6 complete PR production cycles, which were performed in six different dairies located in the PR production area. The relation among microbial dynamics, peptide evolution, and ripening times were investigated in this unique and tightly controlled production and sampling set up. The study of microbial and peptide moieties in products from different dairies – from curd to at least 12 months, the earliest time from which the cheese can be sold, and up to a maximum of 24 months of ripening – highlighted the presence of differences between samples coming from different dairies, probably due to small differences in the cheese making process. Besides these differences, however, ripening time had by far the greatest impact on microbial dynamics and, consequently, on peptide composition.

While durum wheat is a major food source in Mediterranean countries, storage (i.e., gluten) proteins are however responsible for celiac disease (CD), a serious autoimmune disease that occurs in genetically predisposed subjects. Different gluten epitopes—defined as “immunogenic” (IP) and “toxic” (TP) peptides—are involved in the pathology and their content in wheat grain depends on environmental and genetic factors. Detection of IP and TP is not trivial, and no work has been conducted so far to identify the genomic regions associated with their accumulation in wheat. In the present study, a genome-wide association study was performed on a durum wheat collection to identify marker–trait associations (MTAs) between 5730 high quality SNPs and the accumulation of CD-related peptides and gluten protein composition measured in two consecutive cropping seasons (2015/2016 and 2016/2017). High-molecular-weight glutenin subunits (HMW-GS) were more stable between the two years, and differences in total gluten proteins were mainly due to low-molecular-weight glutenin subunits (LMW-GS) and accumulation of gliadins. In the first instance, association tests were conducted on yellow pigment content (YP), a highly inheritable trait with a well-known genetic basis, and several significant MTAs were found corresponding to loci already known for being related to YP. These findings showed that MTAs found for the rest of the measured traits were reliable. In total, 28 significant MTAs were found for gluten composition, while 14 were found to be associated with IP and TP. Noteworthy, neither significant (−log10p > 4.7) nor suggestive (−log10p > 3.3) MTAs for the accumulation of CD-triggering epitopes were found on Gli-A1/Glu-A3 and Gli-B1/Glu-B3 loci, thus suggesting regulatory rather than structural gene effect. A PBF transcription factor on chromosome 5B, known to be involved in the regulation of the expression of CD-related peptides, was identified among the positional candidate genes in the LD-decay range around significant SNPs. Results obtained in the present study provide useful insights and resources for the long-term objective of selecting low-toxic durum wheat varieties while maintaining satisfactory gluten quality. info:eu-repo/semantics/publishedVersion