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Introduction: Fermentation-produced protein (FPP) is gaining global interest as a means of protein production with potentially lower cost and environmental footprint than conventionally-produced animal-sourced proteins. However, estimates on the potential performance of FPP vary substantially, limiting assessment of its scalability and utility. Methods: We integrate life cycle analysis data with nutritional and economic data in an interactive online tool, simulating the requirements and consequences of fermentation at a globally-relevant scale. Results: The tool demonstrates that production of an additional 18 million tons of protein annually via fermentation (~10% of 2020 global consumption) would necessitate 10-25 million hectares of feedstock cropland expansion/reallocation, utilize up to 1% of global electricity generation, produce 159 million tons CO2 equivalents, and have a total process input cost of 53.77 billion USD, with a negligible impact on nutrient supply beyond protein. Discussion: This tool should be used to inform the debate on the future use of fermentation in the food system. [ABSTRACT FROM AUTHOR]

The aquaculture sector faces sustainability challenges due to unsustainable feedstuffs. To address this, there’s a continuous search for alternatives to traditional aquafeed protein sources such as fishmeal. Clostridium autoethanogenum is a novel aquafeed protein source with a long-standing application in the bio-energy industry. With 80–89% crude protein content on a dry matter basis, Clostridium autoethanogenum protein (CAP) has gained attention in nutrition research for aquaculture species. This study aimed to review CAP utilization in aquafeed, examining its nutritional value and impacts on aquatic animals’ growth, feed efficiency, digestion, absorption, and immune response. Research suggests promising potential for CAP in aquafeed, showing potential for achieving complete fishmeal replacement without compromising growth performance and immune response in select aquaculture species. Nonetheless, higher inclusion levels show adverse effects on the health status of certain fish species, indicating variability among species to utilize CAP-based diets. Further investigations are required to understand this variability in outcomes reported among species. [ABSTRACT FROM AUTHOR]

IntroductionFermentation-produced protein (FPP) is gaining global interest as a means of protein production with potentially lower cost and environmental footprint than conventionally-produced animal-sourced proteins. However, estimates on the potential performance of FPP vary substantially, limiting assessment of its scalability and utility.MethodsWe integrate life cycle analysis data with nutritional and economic data in an interactive online tool, simulating the requirements and consequences of fermentation at a globally-relevant scale.ResultsThe tool demonstrates that production of an additional 18 million tons of protein annually via fermentation (~10% of 2020 global consumption) would necessitate 10–25 million hectares of feedstock cropland expansion/reallocation, utilize up to 1% of global electricity generation, produce 159 million tons CO2 equivalents, and have a total process input cost of 53.77 billion USD, with a negligible impact on nutrient supply beyond protein.DiscussionThis tool should be used to inform the debate on the future use of fermentation in the food system.

The search for novel plant proteins presents a major challenge to the global food industry. Beyond the sustainability and nutritional value of seed proteins, there is a growing focus on exploring their potential health benefits for human consumption. However, the endeavour to evaluate the allergenic risks associated with plant-based proteins will require substantial efforts in the coming years. One key strategy to address this concern is the reduction of allergenicity in seeds through the process of germination, which triggers the strong proteolysis of storage proteins. In this study, Lupinus luteus L. seeds were germinated at 25°C for 0, 3, 6 and 9 days under a constant humidity of 80%. Peptide extracts derived from sprouted lupin seeds were subsequently separated under reducing conditions by SDS-PAGE. The resulting protein bands were digested and subjected to mass spectrometry analysis (SWATH-MS) for accurate identification of allergenic proteins. This study's findings revealed the great impact of lupin seed germination on the protein patterns of hydrolysates, particularly noticeable after a germination period of 6 days. The main protein bands (90, 70, 64, 52, 46 and 37 kDa) exhibited a notable decrease in relative intensity as a result of germination. Furthermore, SWATH-MS analysis successfully identified two distinct isoforms (B0YJF7 and B8Q5G0) of the major allergen, β-conglutin, in all tested bands, both prior to and following germination. This study provides evidence for the crucial role of germination in the degradation of this allergen, leading to the elimination of epitopes and subsequently reducing its allergenicity. [ABSTRACT FROM AUTHOR]

Simple Summary: Food proteins from new sources such as vegetable origin (pulses, legumes, cereals), fungi, bacteria and insects are being introduced into the market. However, these novel foods, which had often not been consumed by humans, pose an important risk to public health. The biggest challenge is to ensure food safety by analysing in detail their compositional, nutritional, toxicological and allergenic properties. As a massive preliminary screening, proteomic methods should be employed to search for potential allergens. This review focuses on proteomic and bioinformatic tools for food researchers to identify allergens in novel foods. There is a multitude of highly valuable online tools and protein databases based on sequence alignment, motif identification or 3-D structure predictions. Thus, plant and animal food allergens, including lipid transfer proteins, profilins, seed storage proteins, lactoglobulins, caseins, tropomyosins, parvalbumins and other similar proteins, could be detected in novel food matrices. Furthermore, novel potential allergens could be found for further analysis. This would imply a major simplification. In recent years, novel food is becoming an emerging trend increasingly more demanding in developed countries. Food proteins from vegetables (pulses, legumes, cereals), fungi, bacteria and insects are being researched to introduce them in meat alternatives, beverages, baked products and others. One of the most complex challenges for introducing novel foods on the market is to ensure food safety. New alimentary scenarios drive the detection of novel allergens that need to be identified and quantified with the aim of appropriate labelling. Allergenic reactions are mostly caused by proteins of great abundance in foods, most frequently of small molecular mass, glycosylated, water-soluble and with high stability to proteolysis. The most relevant plant and animal food allergens, such as lipid transfer proteins, profilins, seed storage proteins, lactoglobulins, caseins, tropomyosins and parvalbumins from fruits, vegetables, nuts, milk, eggs, shellfish and fish, have been investigated. New methods for massive screening in search of potential allergens must be developed, particularly concerning protein databases and other online tools. Moreover, several bioinformatic tools based on sequence alignment, motif identification or 3-D structure predictions should be implemented as well. Finally, targeted proteomics will become a powerful technology for the quantification of these hazardous proteins. The ultimate objective is to build an effective and resilient surveillance network with this cutting-edge technology. [ABSTRACT FROM AUTHOR]

New types of protein sources will enter our diet in a near future, reinforcing the need for a straightforward in vitro (cell‐based) screening model to test and predict the safety of these novel proteins, in particular their potential risk for de novo allergic sensitization. The Adverse Outcome Pathway (AOP) for allergen sensitization describes the current knowledge of key events underlying the complex cellular interactions that proceed allergic food sensitization. Currently, there is no consensus on the in vitro model to study the intestinal translocation of proteins as well as the epithelial activation, which comprise the first molecular initiation events (ME1‐3) and the first key event of the AOP, respectively. As members of INFOGEST, we have highlighted several critical features that should be considered for any proposed in vitro model to study epithelial protein transport in the context of allergic sensitization. In addition, we defined which intestinal cell types are indispensable in a consensus model of the first steps of the AOP, and which cell types are optional or desired when there is the possibility to create a more complex cell model. A model of these first key aspects of the AOP can be used to study the gut epithelial translocation behavior of known hypo‐ and hyperallergens, juxtaposed to the transport behavior of novel proteins as a first screen for risk management of dietary proteins. Indeed, this disquisition forms a basis for the development of a future consensus model of the allergic sensitization cascade, comprising also the other key events (KE2‐5). [ABSTRACT FROM AUTHOR]

Due to the increasing global population and climate change, new sustainable food sources are being intensively sought to replace less favorable livestock production. Especially new protein sources and their food applications are being focused on. In this paper, several selected protein sources that may have potential application in future functional foods, such as fermented foods, were examined and compared. These sources include single cell protein (SCP), Arthrospira platensis (Algae), Acheta domesticus (edible insect), potato, and rice protein. The above sources were compared to whey proteins. The parameters studied were total nutritional value, amino acid profile, fatty acid profile, the content of some elements, and the presence of toxins. [ABSTRACT FROM AUTHOR]

Plant-based dairy alternatives have many benefits in terms of sustainability, animal welfare, and health, but they can only be successful in the market if consumers perceive them as suitable substitutes for conventional dairy. Consumers' expectations for new products are strongly influenced by the food categories into which they place these products. The present study aims to reveal consumers' categorizations of plant-based dairy products to gain insights into their potential as dairy substitutes. In a free sorting task, 100 participants from the German-speaking part of Switzerland sorted a variety of plant-based and conventional animal-based dairy products into groups, indicating their spontaneous similarity perceptions. Additionally, we assessed the participants' characteristics and attitudes toward plant-based dairy products to test potential differences in categorization strategies among consumer groups. Multidimensional scaling and cluster analysis showed that consumers' mental representations of plant-based dairy and conventional animal-based dairy were clearly separated across a wide range of product types. This pattern was even observed among consumers who ate less meat, had higher exposure to vegan dietary styles, and had less negative attitudes toward dairy alternatives. The results suggest that taxonomic distinctions based on plant or animal origin dominate consumers' perceptions and are likely to hinder the substitution of dairy with plant-based dairy. Nevertheless, they also imply that plant-based products that manage to emphasize shared goals and functional properties akin to conventional dairy products are more likely to form a common goal-derived category in consumers' minds and thus have better prospects as substitutes., Competing Interests: Declaration of competing interest None., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In recent years, novel food is becoming an emerging trend increasingly more demanding in developed countries. Food proteins from vegetables (pulses, legumes, cereals), fungi, bacteria and insects are being researched to introduce them in meat alternatives, beverages, baked products and others. One of the most complex challenges for introducing novel foods on the market is to ensure food safety. New alimentary scenarios drive the detection of novel allergens that need to be identified and quantified with the aim of appropriate labelling. Allergenic reactions are mostly caused by proteins of great abundance in foods, most frequently of small molecular mass, glycosylated, water-soluble and with high stability to proteolysis. The most relevant plant and animal food allergens, such as lipid transfer proteins, profilins, seed storage proteins, lactoglobulins, caseins, tropomyosins and parvalbumins from fruits, vegetables, nuts, milk, eggs, shellfish and fish, have been investigated. New methods for massive screening in search of potential allergens must be developed, particularly concerning protein databases and other online tools. Moreover, several bioinformatic tools based on sequence alignment, motif identification or 3-D structure predictions should be implemented as well. Finally, targeted proteomics will become a powerful technology for the quantification of these hazardous proteins. The ultimate objective is to build an effective and resilient surveillance network with this cutting-edge technology.

Finding alternative sources of protein is a solution to the shortage of protein caused by societal, environmental, and health concerns. Due to its environmental friendliness, sustainability, and high protein content characteristics, yeast protein (YP) could be a possible alternative protein source. In this paper, the processing properties of YP are examined and contrasted with those of soy protein isolate (SPI), pea protein isolate (PPI), and wheat gluten (WG). The YP was made by fermentation of Saccharomyces cerevisiae , centrifugation to remove nucleic acids, evaporation and concentration, and spray drying. In terms of nutrition, YP had the highest quantity of amino acids (75.91 g/100 g) and included all essential amino acids. It has excellent food processing properties, including foaming (42.5%), emulsification (14.24 m2/g), and thermal stability, but low solubility and fat absorption (0.92 g/g). The current findings highlight that YP could be a promising source of protein for various food applications. [Display omitted] • Functional properties of plant proteins and YP were compared for the first time. • YP contained all essential amino acids and was rich in methionine and lysine. • YP had better foaming and emulsification properties than plant proteins. • There is potential for using YP as a replacement protein. • The study provides theoretical guidance for the application of YP processing. [ABSTRACT FROM AUTHOR]

Simple Summary: Insects are increasingly being used in Europe as a new or alternative source of protein for both direct human consumption and ingredients for feed and food production. Upscaling edible insect production and processing to a sustainable industrial sector is critical to supply the market and meet the foreseen future demands. In a market where transition to more sustainable protein sources is one of the primary challenges, sustainable insect products can contribute to a circular and sustainable economy as well as food security. SUSINCHAIN (SUStainable INsect CHAIN) is a European Horizon 2020 project that aims to contribute to overcome technical and market barriers to enable sustainable large-scale production and consumption of insect proteins in Europe by generating and sharing knowledge, as well as testing, piloting, and demonstrating newly developed insect chain innovations and increasing societal engagement. This article provides an outline of the various obstacles to upscaling of the insect sector and the project's contributions to overcome these. The project covers the topics of: market opportunities, consumer perception, optimization of insect rearing conditions and substrates, insect transportation and processing techniques, application of insect products in food and feed, food safety issues in insect production and processing, together with economic and environmental sustainability. The project's outcomes will provide tools for scaling up and commercializing the European insect sector. The expected global population growth to 9.7 billion people in 2050 and the significant change in global dietary patterns require an increase in global food production by about 60%. The protein supply for feed and food is most critical and requires an extension in protein sources. Edible insects can upgrade low-grade side streams of food production into high-quality protein, amino acids and vitamins in a very efficient way. Insects are considered to be the "missing link" in the food chain of a circular and sustainable economy. Insects and insect-derived products have entered the European market since first being acknowledged as a valuable protein source for feed and food production in around 2010. However, today, scaling up the insect value chain in Europe is progressing at a relatively slow pace. The mission of SUSINCHAIN (SUStainable INsect CHAIN)—a four-year project which has received funding from the European Commission—is to contribute to novel protein provision for feed and food in Europe by overcoming the remaining barriers for increasing the economic viability of the insect value chain and opening markets by combining forces in a comprehensive multi-actor consortium. The overall project objective is to test, pilot and demonstrate recently developed technologies, products and processes, to realize a shift up to Technology Readiness Level 6 or higher. In addition to these crucial activities, the project engages with stakeholders in the insect protein supply chain for feed and food by living labs and workshops. These actions provide the necessary knowledge and data for actors in the insect value chain to decrease the cost price of insect products, process insects more efficiently and market insect protein applications in animal feed and regular human diets that are safe and sustainable. This paves the way for further upscaling and commercialization of the European insect sector. [ABSTRACT FROM AUTHOR]

Protein transition, i.e. the transition from high levels of traditional meat consumption towards consuming less meat or more plant-based or alternative animal-based proteins, is highly dependent on consumer behaviour. This position paper adds to the literature by integrating the research streams on behavioural sciences and meat reducing strategies, thereby contributing to the use of behavioural science insights in developing meat reducing interventions towards a more plant-based food transition. Meat-reducing strategies involve substituting meat with novel proteins, consumption of less meat or consuming meat less often, and becoming a vegetarian or vegan. Based on previous literature four systematic steps for effective interventions towards behaviour change are described in view of the current literature in the specific context of meat reduction. Finally, emergent strands of future research are identified. The four described steps compromise: (a) identifying the problem and desired behaviour change, (b) examining the main drivers of behaviour change, (c) select fitting interventions, and (d) impact assessment. Based on the meat-reducing literature the key strands for future interventions in the context of protein transition are identified. Moreover, literature gaps are defined. Resulting in an overview of systematic steps for interventions to support behaviour change in the protein transition. • Reaching behaviour change towards less traditional meat consumption is difficult. • Social sciences can support development of interventions towards meat reducing behaviour. • Four steps: behaviour identification, understanding of drivers, intervention selection, and assessing impact. • Findings can be used to select, design and implement interventions to support protein transition. [ABSTRACT FROM AUTHOR]

Due to the increasing global population and climate change, new sustainable food sources are being intensively sought to replace less favorable livestock production. Especially new protein sources and their food applications are being focused on. In this paper, several selected protein sources that may have potential application in future functional foods, such as fermented foods, were examined and compared. These sources include single cell protein (SCP), Arthrospira platensis (Algae), Acheta domesticus (edible insect), potato, and rice protein. The above sources were compared to whey proteins. The parameters studied were total nutritional value, amino acid profile, fatty acid profile, the content of some elements, and the presence of toxins.

One of the technological responses to concern about the healthiness and sustainability of red meat consumption as well as growing global food insecurity has been the development of plant-based meats. While still niche industries, plant-based meats are increasing in diversity and visibility and are often accompanied by scientific claims about their potential contribution to sustainability, health and nutrition. In this paper we analyse these claims from the perspective of critical metrology, looking not only at the state of knowledge about plant- and animal-based beef but also how knowledge is being produced. Such research is vital in understanding not only what is known about a topic but also the limitations of that knowledge in terms of influences and gaps, to shape how research can be done better. The research is based on a scoping review of scientific papers that compare the sustainability and nutritional properties of plant- and animal-based beef, with a particular focus on the Australian context. Our analysis shows that there are some points of consensus, such as plant-based beef having lower greenhouse gas emissions than animal-based beef and that plant-based burgers have lower total fat and saturated fat than animal-based burgers. However, we also find points of difference due to variability in production systems, products and approaches to measurement. We discuss how a metrological regime is forming that bears the imprint and interests of the industries involved, influencing the metrics and geographies that are studied and creating critical absences in understanding. [ABSTRACT FROM AUTHOR]

The expected global population growth to 9.7 billion people in 2050 and the significant change in global dietary patterns require an increase in global food production by about 60%. The protein supply for feed and food is most critical and requires an extension in protein sources. Edible insects can upgrade low-grade side streams of food production into high-quality protein, amino acids and vitamins in a very efficient way. Insects are considered to be the “missing link” in the food chain of a circular and sustainable economy. Insects and insect-derived products have entered the European market since first being acknowledged as a valuable protein source for feed and food production in around 2010. However, today, scaling up the insect value chain in Europe is progressing at a relatively slow pace. The mission of SUSINCHAIN (SUStainable INsect CHAIN)—a four-year project which has received funding from the European Commission—is to contribute to novel protein provision for feed and food in Europe by overcoming the remaining barriers for increasing the economic viability of the insect value chain and opening markets by combining forces in a comprehensive multi-actor consortium. The overall project objective is to test, pilot and demonstrate recently developed technologies, products and processes, to realize a shift up to Technology Readiness Level 6 or higher. In addition to these crucial activities, the project engages with stakeholders in the insect protein supply chain for feed and food by living labs and workshops. These actions provide the necessary knowledge and data for actors in the insect value chain to decrease the cost price of insect products, process insects more efficiently and market insect protein applications in animal feed and regular human diets that are safe and sustainable. This paves the way for further upscaling and commercialization of the European insect sector.

In recent years, proteogenomics and ribosome profiling studies have identified a large number of proteins encoded by noncoding regions in the human genome. They are encoded by small open reading frames (sORFs) in the untranslated regions (UTRs) of mRNAs and long non-coding RNAs (lncRNAs). These sORF encoded proteins (SEPs) are often <150AA and show poor evolutionary conservation. A subset of them have been functionally characterized and shown to play an important role in fundamental biological processes including cardiac and muscle function, DNA repair, embryonic development and various human diseases. How many novel protein-coding regions exist in the human genome and what fraction of them are functionally important remains a mystery. In this review, we discuss current progress in unraveling SEPs, approaches used for their identification, their limitations and reliability of these identifications. We also discuss functionally characterized SEPs and their involvement in various biological processes and diseases. Lastly, we provide insights into their distinctive features compared to canonical proteins and challenges associated with annotating these in protein reference databases. • Comprehensive review of sORF encoded proteins (SEPs) encoded by the human genome. • Experimentally verified SEPs, their potential function and disease association. • Difficulties in identifying and functionally characterizing SEPs. • Some SEPs could be translational noise. [ABSTRACT FROM AUTHOR]

Current international guidelines for the risk assessment of biotechnology-derived foods date back to 2003. We present new strategies and directions for assessing immune adverse reactions to novel food proteins. Understanding genetic factors involved in food allergy and the role of the gastrointestinal tract will streamline risk assessment strategies. [ABSTRACT FROM AUTHOR]

Proteins are among the most important constituents of biological systems. Because all protein-coding genes have a noncoding ancestral form, the properties of noncoding sequences and how they shape the birth of novel proteins may influence the structure and function of all proteins. Differences between the properties of young proteins and random expectations from noncoding sequences have previously been interpreted as the result of natural selection. However, interpreting such deviations requires a yetunattained understanding of the raw material of de novo gene birth and its relation to novel functional proteins. We mathematically show that the average properties and selective filtering of the "junk" polypeptides of which this raw material is composed are not the only factors influencing the properties of novel functional proteins. We find that in some biological scenarios, they also depend on the variance of the properties of junk polypeptides and their correlation with the rate of allelic turnover, which may itself depend on mutational biases. This suggests for instance that any property of polypeptides that accelerates their exploration of the sequence space could be overrepresented in novel functional proteins, even if it has a limited effect on adaptive value. To exemplify the use of our general theoretical results, we build a simple model that predicts the mean length and mean intrinsic disorder of novel functional proteins from the genomic GC content and a single evolutionary parameter. This work provides a theoretical framework that can guide the prediction and interpretation of results when studying the de novo emergence of protein-coding genes. [ABSTRACT FROM AUTHOR]

The current allergenicity assessment of novel proteins is based on the EFSA GMO guidance. Recently, EFSA launched a new guidance document on allergenicity assessment of GM plants (2017). This document describes, amongst other topics, the new scientific and regulatory developments on in vitro protein digestibility tests. The EFSA GMO Panel stated that for in vitro protein digestibility tests, additional investigations are needed before any additional recommendation in the form of guidance can be provided. To this end, an interim phase is considered necessary to evaluate the revisions to the in vitro gastrointestinal digestion test, proposed by EFSA. This prompted the establishment of a joint workshop through two COST Action networks: COST Action ImpARAS and COST Acton INFOGEST. In 2017, a workshop was organised to discuss the relevance of digestion in allergenicity risk assessment and how to potentially improve the current methods and readouts. The outcome of the workshop is that there is no rationale for a clear readout that is predictive for allergenicity and we suggest to omit the digestion test from the allergenicity assessment strategy for now, and put an effort into filling the knowledge gaps as summarized in this paper first. • Protein digestion is relevant for allergenicity of some proteins, but not for all. • It is not clear yet how different factors exactly influence digestion and how these factors can be included. • Including all factors will complicate the elaboration of a simple and predictive digestion assay. • There is no rationale for a clear readout that is predictive for allergenicity. • We suggest to omit the digestion test from the allergenicity assessment for now and put effort into filling the knowledge gaps. [ABSTRACT FROM AUTHOR]

Protein hydrolysates from lesser mealworm (Alphitobius diaperinus, LM) were obtained by enzymatic hydrolysis with protease from Bacillus licheniformis. A preliminary test performed for five hours of hydrolysis generated an insect protein hydrolysate with 15% of degree of hydrolysis (DH), optimum solubility property and oil holding capacity, but emulsifying and foaming ability were completely impaired. In order to investigate the potential implication of DH on techno-functional properties, a set of protein hydrolysates with a different DH was obtained by sub-sampling at different time points during three hours of enzymatic hydrolysis process. An increase in DH% had positive effects on the solubility property and oil holding ability, while a reduced emulsifying ability was observed up to five hours of hydrolysis. These results demonstrated that the enzymatic hydrolysis, if performed under controlled conditions and not for a long period, represents a valid method to extract high quality protein from insects with tailored techno-functionality, in order to produce tailored ingredients for feed and food purpose.

The development and introduction of new dietary protein sources has the potential to improve food supply sustainability. Understanding the potential allergenicity of these new or modified proteins is crucial to ensure protection of public health. Exposure to new proteins may result in de novo sensitization, with or without clinical allergy, or clinical reactions through cross-reactivity. In this paper we review the potential of current methodologies ( in silico , in vitro degradation, in vitro IgE binding, animal models and clinical studies) to address these outcomes for risk assessment purposes for new proteins, and especially to identify and characterise the risk of sensitization for IgE mediated allergy from oral exposure. Existing tools and tests are capable of assessing potential crossreactivity. However, there are few possibilities to assess the hazard due to de novo sensitization. The only methods available are in vivo models, but many limitations exist to use them for assessing risk. We conclude that there is a need to understand which criteria adequately define allergenicity for risk assessment purposes, and from these criteria develop a more suitable battery of tests to distinguish between proteins of high and low allergenicity, which can then be applied to assess new proteins with unknown risks. [ABSTRACT FROM AUTHOR]

Currently, risk assessment of the allergenic potential of novel proteins relies heavily on evaluating protein digestibility under normal conditions based on the theory that allergens are more resistant to gastrointestinal digestion than non-allergens. There is also proposed guidance for expanded in vitro digestibility assay conditions to include vulnerable sub-populations. One of the underlying rationales for the expanded guidance is that current in vitro assays do not accurately replicate the range of physiological conditions. Animal scientists have long sought to predict protein and amino acid digestibility for precision nutrition. Monogastric production animals, especially swine, have gastrointestinal systems similar to humans, and evaluating potential allergen digestibility in this context may be beneficial. Currently, there is no compelling evidence that the mechanisms sometimes postulated to be associated with allergenic sensitization, e.g. antacid modification of stomach pH, are valid among production animals. Furthermore, examples are provided where non-biologically representative assays are better at predicting protein and amino acid digestibility compared with those designed to mimic in vivo conditions. Greater emphasis should be made to align in vitro assessments with in vivo data. [ABSTRACT FROM AUTHOR]

Replacement of current protein resources for novel or alternative sources may be one of the solutions to abolish the expected scarcity of dietary protein for animal feeds. However, little is known about the nutritional, protein composition, and potential functional value of such novel or alternative protein sources. In the present study, we used advanced proteomic and bioinformatic approaches to characterize the protein component of 6 different protein sources and predicted the biofunctionalities of their in vivo digestion products. We used casein (CAS), partially delactosed whey powder (DWP), spray-dried porcine plasma (SDPP), soybean meal (SBM), wheat gluten meal (WGM), and yellow meal worm (YMW). We characterized and semiquantifed the individual proteins present in these resources by nano-liquid chromatography-LTQ Orbitrap-mass spectrometry (nLCMS). Based on the data obtained, we calculated the AA composition of the proteins that constitute 90% of the total calculated protein content and compared this with the chemically determined AA compositions of the corresponding protein sources. By the use of bioinformatic procedures, we predicted the bioactive properties of these protein sources after in silico digestion with monogastric proteolytic enzymes. We detected and semiquantifed 37, 58, 85, 188, 113, and 33 different individual proteins in CAS, DWP, SDPP, SBM, WGM, and YMW, respectively. The calculated AA composition of the various protein sources was almost identical to the chemically determined composition, with correlation values ranging from 0.85 to 0.94. Furthermore, we revealed that the selected protein sources are potentially rich in bioactive peptides, in particular of angiotensin-converting enzyme inhibitors and peptides with antioxidative properties. We discuss the results in terms of the beneft of the applied nLCMSbased approach for analyzing protein feed ingredients and the use of these alternative sources of protein in animal feeds for monogastrics. Furthermore, we discuss new potential applications of this method in the area of (animal) nutrition. [ABSTRACT FROM AUTHOR]

Alternative protein sources are being incorporated in companion animal diets as a response to the market demand for high quality products. Chicken is a commonly used protein in pet food and was used as a comparison for emerging alternatives goose, quail, duck, and emu which have been evaluated in the current study. All protein sources were analyzed for chemical composition, and amino acid (AA) standardized digestibility and calculation of digestible indispensable amino acid scores (DIAAS-like) were determined by using the precision fed cecectomized rooster assay. Dry matter values ranged from 95% to 98% and were similar for all proteins. Crude protein was least for goose meat and greatest in emu meat (42% and 95%, respectively). Emu meat produced the least fat content while quail produced the most (4.7% and 42%, respectively). Standardized AA digestibility values for the novel proteins were above 60% for indispensable AA. The DIAAS-like calculation applied reference proteins based on Association of American Feed Control officials (AAFCO) recommended nutrient profile and National Research Council (NRC) recommended allowances for adult dogs and cats at maintenance. The DIAAS-like values for cats for AAFCO and NRC were >100%. Tryptophan was identified to be the firstlimiting AA in every protein source except for goose meat where methionine was first-limiting using NRC recommended values for adult dogs. [ABSTRACT FROM AUTHOR]

There has been a quest for novel and alternative protein sources in pet nutrition due to product diversification, need for novel and single protein diets, and concerns with sustainability. However, limited information on chemical composition and amino acid (AA) digestibility of these ingredients are available. The novel protein sources tested are of aquatic and reptile species, and include fish slurry, white fish, spirulina, eel, salmon, and rattlesnake. We determined the chemical composition and applied the cecectomized rooster model to estimate standardized AA digestibility of these protein sources. The ingredients were further assessed by calculation of digestible indispensable amino acid scores (DIAAS-like). Dry matter values were similar across all proteins and ranged from 91.3% to 99.4%. Crude protein was least in salmon (43.5%) and greatest in fish slurry and spirulina (72.5% and 72.8%, respectively). Rattlesnake produced the least amount of total fat at 1.8% and eel produced the most at 32.3%. Standardized AA values were >80% for both dispensable and indispensable AA in all protein sources. For DIAAS-like calculations, reference proteins were based on nutrient profiles of the Association of American Feed Control officials (AAFCO) nutrient profile and the National Research Council (NRC) recommended allowances for adult dogs and cats at maintenance. Using these references, rattlesnake was low quality for both dogs and cats. Fish slurry and spirulina were observed to be high quality for dogs and cats using AAFCO references and high quality in only cats using NRC references. Using the NRC reference for dogs, fish slurry and spirulina were determined to be moderate quality because the first-limiting AA in fish slurry (tryptophan) was 78% and first-limiting AA in spirulina (methionine) was 63%. Scores < 100% but >50% are considered moderate quality according to the DIAAS method. [ABSTRACT FROM AUTHOR]

Ribosome profiling has revealed translation outside canonical coding sequences, including translation of short upstream ORFs, long noncoding RNAs, overlapping ORFs, ORFs in UTRs, or ORFs in alternative reading frames. Studies combining mass spectrometry, ribosome profiling, and CRISPR-based screens showed that hundreds of ORFs derived from noncoding transcripts produce (micro)proteins, whereas other studies failed to find evidence for such types of noncanonical translation products. Here, we attempted to discover translation products from noncoding regions by strongly reducing the complexity of the sample prior to mass spectrometric analysis. We used an extended database as the search space and applied stringent filtering of the identified peptides to find evidence for novel translation events. We show that, theoretically our strategy facilitates the detection of translation events of transcripts from noncoding regions but experimentally only find 19 peptides that might originate from such translation events. Finally, Virotrap-based interactome analysis of two N-terminal proteoforms originating from noncoding regions showed the functional potential of these novel proteins., Competing Interests: Conflict of interest The authors declare no competing interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Development of genetically modified (GM) crops is on increase to improve food quality, increase harvest yields, and reduce the dependency on chemical pesticides. Before their release in marketplace, they should be scrutinized for their safety. Several guidelines of different regulatory agencies like ILSI, WHO Codex, OECD, and so on for allergenicity evaluation of transgenics are available and sequence homology analysis is the first test to determine the allergenic potential of inserted proteins. Therefore, to test and validate, 312 allergenic, 100 non-allergenic, and 48 inserted proteins were assessed for sequence similarity using 8-mer, 80-mer, and full FASTA search. On performing sequence homology studies, ~94% the allergenic proteins gave exact matches for 8-mer and 80-mer homology. However, 20 allergenic proteins showed non-allergenic behavior. Out of 100 non-allergenic proteins, seven qualified as allergens. None of the inserted proteins demonstrated allergenic behavior. In order to improve the predictability, proteins showing anomalous behavior were tested by Algpred and ADFS separately. Use of Algpred and ADFS softwares reduced the tendency of false prediction to a great extent (74--78%). In conclusion, routine sequence homology needs to be coupled with some other bioinformatic method like ADFS/Algpred to reduce false allergenicity prediction of novel proteins. [ABSTRACT FROM AUTHOR]

Background/Aims: It is still unclear what happens in the glomerulus when proteinuria starts. Using puromycin aminonucleoside nephrosis (PAN) rats, we studied early ultrastructural and permeability changes in relation to the expression of the podocyte-associated molecules nephrin, α-actinin, dendrin, and plekhh2, the last two of which were only recently discovered in podocytes. Methods: Using immune stainings, semiquantitative measurement was performed under the electron microscope. Permeability was assessed using isolated kidney perfusion with tracers. Possible effects of ACE inhibition were tested. Results: By day 2, some patchy foot process effacement, but no proteinuria, appeared. The amount of nephrin was reduced in both diseased and normal areas. The other proteins showed few changes, which were limited to diseased areas. By day 4, foot process effacement was complete and proteinuria appeared in parallel with signs of size barrier damage. Nephrin decreased further, while dendrin and plekhh2 also decreased but α-actinin remained unchanged. ACE inhibition had no significant protective effect. Conclusions: PAN glomeruli already showed significant pathology by day 4, despite relatively mild proteinuria. This was preceded by altered nephrin expression, supporting its pivotal role in podocyte morphology. The novel proteins dendrin and plekhh2 were both reduced, suggesting roles in PAN, whereas α-actinin was unchanged. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Abstract: Comparative proteomics analysis of the cytosolic proteins of Salmonella Gallinarum (SG) and Salmonella Enteritidis (SE) isolated from poultry was performed. The constantly detected spots of serovar SG with concomitant absence in SE serovar as well as those markedly over expressed in serovar SE were selected for MALDI-TOF-MS identification. The NCBI-matched proteins that show overregulation were then further confirmed on the mRNA level by quantitative real time PCR. Identified proteins were representing diverse functional activities including energy production, metabolism, and nucleic acid synthesis. Interestingly, some recognized proteins have some relevance to bacterial virulence e.g. Salmonella pathogenicity island 1 effector protein, T-cell inhibitor protein, response regulator protein, paratose synthetase protein (RfbS) and heat shock protein 90. The study revealed the presence of some proteins of unknown function, which raise the speculation for their importance in either host adaptation or pathogenicity among SG serovars. [Copyright &y& Elsevier]

Industrial valorization of proteins from traditional edible insects requires knowledge of their nutritional, chemical, technological characteristics. Therefore, these characteristics in two Mexican edible insects (A. cordifera and B. mellifica) were studied and compared to commercial insect species (H. illucens). The nutritional attributes of insect protein concentrates (PCs) were determined by their protein content, amino acid and polyphenol compositions. The chemical structure of the proteins was determined by Fourier-Transform Infrared spectroscopy (FTIR) and Differential Scanning Calorimetry (DSC). Moreover, the surfactant properties of insect-PC (emulsions) were evaluated. B. mellifica PC showed the highest content of protein content among the studied insects (57.1%). In contrast, H. illucens showed the highest polyphenols (12.7 mg/g). The emulsions prepared with insect-PC from A. cordifera and B. mellifica showed higher stability than H. illucens , where a bigger droplet size and a rapid phase separation occurred. The decrease in emulsification properties of black soldier fly larvae was attributed to a possible interaction between polyphenols and proteins that hinders emulsification. This study increases the understanding of the structural and techno-functional characteristics of insect-PC of commercial and local insect species. • Protein concentrates of jumil and wasp have higher emulsion capacity. • BSF protein concentrate is abundant in polyphenols, mainly epicatechin gallate. • Polyphenols in BSF protein concentrate could hinder its emulsion capacity. [ABSTRACT FROM AUTHOR]

Abstract: There is a need for novel protein sources. Insects are a possible interesting source of protein. They are nutritious in terms of protein (40–75 g/100g dry weight) and minerals. Insect protein is of high quality and has a high digestibility (77–98%) and concentration of essential amino acids (46–96% of the nutritional profile). Also insect cells may be a promising novel source of protein. Choice of cell line, growth conditions and use of the baculovirus expression system opens up possibilities to engineer the nutritional value of the biomass. The technological limits as well as consumer acceptance of insect cell based food remains to be investigated. [Copyright &y& Elsevier]

Glue proteins are tissue-specific proteins synthesized by larval salivary gland cells of Drosophila. In Drosophila nasuta nasuta and D. n. albomicans of the nasuta subgroup, the genes that encode the major glue protein fractions are X-linked. In the present study, these X-linked markers have been employed to trace the pattern of introgression of D. n. nasuta and D. n. albomicans genomes with respect to the major glue protein fractions in their interracial hybrids, called cytoraces. These cytoraces have inherited the chromosomes of both parents and have been maintained in the laboratory for over 400-550 generations. The analysis has revealed that cytoraces with D. n. albomicans X chromosome show either D. n. nasuta pattern or a completely novel pattern of glue protein fractions. Further, quantitative analysis also shows lack of correlation between the chromosomal pattern of inheritance and overall quantity of the major glue protein fractions in the cytoraces. Thus, in cytoraces the parental chromosomes are not just differentially represented but there is evidence for introgression even at the gene level. [ABSTRACT FROM AUTHOR]

Abstract Mitochondria of cytoplasmic male sterile crop plants contain novel, chimeric open reading frames. In addition, a number of crops carry endogenous double-stranded ribonucleic acid (dsRNA). In this study, the novel proteins encoded by these genetic components were screened for the presence of potential binding sites (epitopes) of allergy-associated IgE antibodies, as was previously done with transgenic proteins from genetically modified crops. The procedure entails the identification of stretches of at least six contiguous amino acids that are shared by novel proteins and known allergenic proteins. These stretches are further checked for potential linear IgE-binding epitopes. Of the 16 novel protein sequences screened in this study, nine contained stretches of six or seven amino acids that were also present in allergenic proteins. Four cases of similarity are of special interest, given the predicted antigenicity of the identical stretch within the allergenic and novel protein, the IgE-binding by a peptide containing an identical stretch reported in literature, or the multiple incidence of identical stretches of the same allergen within a novel protein. These selected stretches are present in novel proteins derived from oilseed rape and radish (ORF138), rice (dsRNA), and fava bean (dsRNA), and warrant further clinical testing. The frequency of positive outcomes and the sizes of the identical stretches were comparable to those previously found for transgenic proteins in genetically modified crops. It is discussed whether novel proteins from conventional crops should be subject to an assessment of potential allergenicity, a procedure which is currently mandatory for transgenic proteins from genetically modified crops. [ABSTRACT FROM AUTHOR]