Non-animal proteins as cutting-edge ingredients to reformulate animal-free foodstuffs: Present status and future perspectives.

Consumer interest in protein rich diets is increasing, with more attention being paid to the protein source. Despite the occurrence of animal proteins in the human diet, non-animal proteins are gaining popularity around the world due to their health benefits, environmental sustainability, and ethical merit. These sources of protein qualify for vegan, vegetarian, and flexitarian diets. Non-animal proteins are versatile, derived mainly from cereals, vegetables, pulses, algae (seaweed and microalgae), fungi, and bacteria. This review's intent is to analyze the current and future direction of research and innovation in non-animal proteins, and to elucidate the extent (limitations and opportunities) of their applications in food and beverage industries. Prior knowledge provided relevant information on protein features (processing, structure, and techno-functionality) with particular focus on those derived from soy and wheat. In the current food landscape, beyond conventionally used plant sources, other plant proteins are gaining traction as alternative ingredients to formulate animal-free foodstuffs (e.g., meat alternatives, beverages, baked products, snack foods, and others). Microbial proteins derived from fungi and algae are also food ingredients of interest due to their high protein quantity and quality, however there is no commercial food application for bacterial protein yet. In the future, key points to consider are the importance of strain/variety selection, advances in extraction technologies, toxicity assessment, and how this source can be used to create food products for personalized nutrition.

Development and Biomass Composition of Ephestia kuehniella (Lepidoptera: Pyralidae), Tenebrio molitor (Coleoptera: Tenebrionidae), and Hermetia illucens (Diptera: Stratiomyidae) Reared on Different Byproducts of the Agri-Food Industry.

The aim of this study was to evaluate five agro-industrial byproducts (apricots, brewer's spent grains, brewer's spent yeast, feed mill byproducts including broken cereal grains, and hatchery waste including eggshell debris, fluff, infertile eggs, dead embryos, and egg fluids) or mixtures thereof as food diets of Ephestia kuehniella (Zeller), Tenebrio molitor (L.), and Hermetia illucens (L.). Eleven out of 26 tested combinations allowed the first instar larvae to reach the adult stage. Results showed that bioconversion parameters and biomass composition can vary depending on the diet composition, especially in the case of E. kuehniella and H. illucens, whose nutritional requirements seem more complex than those of T. molitor. Tenebrio molitor was able to develop in almost all byproducts. However, only when T. molitor was fed with suitable mixtures of byproducts the development parameters were similar to those obtained with the standard diet. The best results in terms of bioconversion parameters were obtained by feeding H. illucens with a diet including dried brewer's spent grain, feed mill byproducts and brewer's spent yeast. The larvae of these three species can be considered interesting from a nutritional point of view, because of their high protein and fat content. However, the fatty acids profile of H. illucens larvae, with high proportions of saturated fatty acids, seems less healthy for human consumption compared with those of E. kuehniella and T. molitor.

Nutritional properties of organic and conventional beef meat at retail.

BACKGROUND: Consumers perceive that organic meat has superior nutritional properties compared to conventional meat, although the available evidence from commercial samples is very scarce. The present study compared the nutritional composition of organic and conventional beef meat sold at retail, including, for the first time, the bioactive compounds coenzyme Q10 , carnosine, anserine, creatine and taurine. Sampling comprised two muscles: longissimus thoracis and supraspinatus. RESULTS: Organic beef had 17% less cholesterol, 32% less fat, 16% less fatty acids, 24% less monounsaturated fatty acids, 170% more α-linolenic acid, 24% more α-tocopherol, 53% more ß-carotene, 34% more coenzyme Q10 and 72% more taurine than conventional beef. Differences between organic and conventional samples were clearly dependent on the muscle because longissimus thoracis and supraspinatus showed different patterns of compound accumulation. CONCLUSION: To our knowledge, a higher amount of bioactive compounds in organic beef meat is reported for the first time in the present study. Retail organic beef had a higher nutritional value than retail conventional beef, which resulted from better-balanced lipid and bioactive compound contents. © 2019 Society of Chemical Industry.

Effect of pH and temperature on tropane alkaloids within a processing strategy to provide safe infant cereal-based food.

Tropane alkaloids (TAs) are secondary metabolites from weeds that can contaminate cereals and vegetables during harvest. Due to their toxicity, the Regulation (EC) 2023/915 sets maximum levels for atropine and scopolamine in cereal-based foods for infants containing millet, sorghum, buckwheat or their derived products. The aim of this study was to evaluate the effect of pH and temperature on the stability of TAs, as possible parameters in thermal processing to mitigate this chemical hazard in cereal-based infant food. The effect of pH (4 and 7) and temperature (80 °C and 100 °C) was assessed in buffer solutions. Also, treatment at 180 °C was performed in spiked and naturally incurred millet flour to assess the effect of high temperature, simulating cooking or drying, on the stability of TAs in the cereal matrix. The fate of 24 TAs was assessed by UHPLC-MS/MS. TAs showed high thermostability, although it was variable depending on the specific compound, pH, temperature and treatment time. In buffer solutions, higher degradation was found at 100 °C and pH 7. In spiked millet flour at 180 °C for 10 min, scopolamine and atropine contents decreased by 25 % and 22 %, similarly to other TAs which also showed a slow thermal degradation. Atropine, scopolamine, anisodamine, norscopolamine, scopine and scopoline were found in naturally contaminated millet flour. Interestingly, naturally incurred atropine was more thermostable than when spiked, showing a protective effect of the cereal matrix on TAs degradation. The present results highlight the need for an accurate monitorization of TAs in raw materials, as this chemical hazard may remain in infant cereal-based food even after intense thermal processing.

Pulse Spray Drying for Bovine Skimmed Milk Powder Production.

Pulse Spray Drying (PSD) has potential as a sustainable means of skimmed milk powder (SMP) production. In this study, powders were obtained from PSD using different drying outlet temperatures (70, 80, 90 and 100 °C), and their characteristics were compared to those of traditional Spray Drying (SD). Native whey proteins were well preserved and Solubility Indexes were over 98% in all cases, despite powders obtained by PSD displaying lower solubility than the SD ones. No visual difference was observable in the powders (ΔE < 2); however, PSD powders were found to be yellower with a higher Browning Index. The drying technology did not have a significant effect on powder moisture content and bulk density. Particle size distribution and scanning electron microscopy images confirmed the presence of fine particles (<10 µm) in all samples that might have provided poor flowability and wetting behavior (overall Carr Index and Hausner ratio were 33.86 ± 3.25% and 1.52 ± 0.07, respectively). Higher amounts of agglomerated particles were found at low temperatures in the products processed with both technologies, but PSD samples showed a narrower particle size distribution and hollow particles with more wrinkles on the surface (probably due to the fast evaporation rate in PSD). Overall, PSD provided SMP with comparable physicochemical characteristics to SD and, once optimized at the industrial level, could offer significant advantages in terms of thermal efficiency without significant modification of the final product quality.

Separation of α-Lactalbumin Enriched Fraction from Bovine Native Whey Concentrate by Combining Membrane and High-Pressure Processing.

Whey exhibits interesting nutritional properties, but its high ß-Lactoglobulin (ß-Lg) content could be a concern in infant food applications. In this study, high-pressure processing (HPP) was assessed as a ß-Lg removal strategy to generate an enriched α-Lactalbumin (α-La) fraction from bovine native whey concentrate. Different HPP treatment parameters were considered: initial pH (physiological and acidified), sample temperature (7-35 °C), pressure (0-600 MPa) and processing time (0-490 s). The conditions providing the best α-La yield and α-La purification degree balance (46.16% and 80.21%, respectively) were 4 min (600 MPa, 23 °C), despite the significant decrease of the surface hydrophobicity and the total thiol content indexes in the α-La-enriched fraction. Under our working conditions, the general effects of HPP on α-La and ß-Lg agreed with results reported in other studies of cow milk or whey. Notwithstanding, our results also indicated that the use of native whey concentrate could improve the ß-Lg precipitation degree and the α-La purification degree, in comparison to raw milk or whey. Future studies should include further characterization of the α-La-enriched fraction and the implementation of membrane concentration and HPP treatment to valorize cheese whey.

Separation of α-Lactalbumin-Enriched Fractions from Caprine and Ovine Native Whey Concentrate by Combining Membrane and High-Pressure Processing.

Whey from goat and sheep have been gaining attention in the last few years for their nutritional properties. Unfortunately, ß-Lg, not found in human milk, may trigger infant allergies if used in infant food formulations, so there is a growing interest in developing ingredients derived from whey with higher α-La/ß-Lg ratios. The objective of this work was to study the effect of high-pressure processing (HPP) on caprine and ovine native whey concentrates (NWC) in order to obtain α-Lactalbumin (α-La)-enriched fractions. NWCs were treated at 600 MPa (23 °C) for 2, 4, and 15 min and two pH conditions were studied (physiological pH and pH 4.60). The concentration of ß-Lg in supernatant fraction after HPP significantly decreased after 2 min of treatment, while the concentration of α-La was unchanged in both goat and sheep samples. Longer HPP processing times (up to 15 min) progressively increased α-La purification degree but also decreased the α-La yield. Caprine and ovine NWCs treated at physiological pH provided better α-La yield, α-La purification degree, and higher ß-Lg precipitation degrees than the corresponding acidified samples, while the corresponding NWC supernatant (NWCsup) showed lower values for both surface hydrophobicity and total free thiol indices, suggesting a higher extent of protein aggregation. Effects of sample acidification and the HPP treatment were opposite to those previously reported on bovine NWC, so further characterization of caprine and ovine ß-Lg should be carried out to understand their different behavior.

Fermentation for Designing Innovative Plant-Based Meat and Dairy Alternatives.

Fermentation was traditionally used all over the world, having the preservation of plant and animal foods as a primary role. Owing to the rise of dairy and meat alternatives, fermentation is booming as an effective technology to improve the sensory, nutritional, and functional profiles of the new generation of plant-based products. This article intends to review the market landscape of fermented plant-based products with a focus on dairy and meat alternatives. Fermentation contributes to improving the organoleptic properties and nutritional profile of dairy and meat alternatives. Precision fermentation provides more opportunities for plant-based meat and dairy manufacturers to deliver a meat/dairy-like experience. Seizing the opportunities that the progress of digitalization is offering would boost the production of high-value ingredients such as enzymes, fats, proteins, and vitamins. Innovative technologies such as 3D printing could be an effective post-processing solution following fermentation in order to mimic the structure and texture of conventional products.

Protein Quality and Protein Digestibility of Vegetable Creams Reformulated with Microalgae Inclusion.

Microalgae are considered a valuable source of proteins that are used to enhance the nutritional value of foods. In this study, a standard vegetable cream recipe was reformulated through the addition of single-cell ingredients from Arthrospira platensis (spirulina), Chlorella vulgaris, Tetraselmis chui, or Nannochloropsis oceanica at two levels of addition (1.5% and 3.0%). The impact of microalgae species and an addition level on the amino acid profile and protein in vitro digestibility of the vegetable creams was investigated. The addition of microalgae to vegetable creams improved the protein content and the amino acid nutritional profile of vegetable creams, whereas no significant differences were observed in protein digestibility, regardless of the species and level of addition, indicating a similar degree of protein digestibility in microalgae species despite differences in their protein content and amino acid profile. This study indicates that the incorporation of microalgae is a feasible strategy to increase the protein content and nutritional quality of foods.

Algae as Nutritional and Functional Food Sources.

Algae are a large and diverse group of autotrophic eukaryotic and photosynthetic aquatic organisms [...].

Levels of taurine, hypotaurine and homotaurine, and amino acids profiles in selected commercial seaweeds, microalgae, and algae-enriched food products.

Amino acids and sulfonic acid derivatives (Taurine-Tau; Hypotaurine-HypTau; Homotaurine-HTau) of 26 different species of commercial macroalgae, microalgae and 10 algae-enriched food products from the market were quantified in a single chromatographic run. Tau and analogues were predominantly distributed in red species followed by green and brown species. Palmaria palmata, Gracilaria longissima and Porphyra sp. were the species with the highest content of Tau and total sulfonic acid derivatives (TAD). Notwithstanding, relatively high concentrations of HTau were found in green algae Ulva lactuca and G. vermicullophyla as well as in the brown algae Undaria pinnatifida. HTau and HypTau were found at lower concentrations than Tau in all species, except in Ulva lactuca. The samples with the highest protein content were the green species Chlorella vulgaris, Nannochloropsis, and Afanizomenon-flos aquae, followed by the red algae Gracilaria longissima and Gracilaria vermicullophyla. Samples of pasta formulated with algae ingredients contained the highest levels of sulfonic acid derivatives, evidencing that these products can provide levels of TAD comparable to those found in foods of animal origin. This study provides, for the first time, quantitative information regarding the distribution of sulfonic acid derivatives and total amino acids in multiple algae species as well as the nutritional impact of the inclusion of algae ingredients in commercial food matrices.

Seafood alternatives: assessing the nutritional profile of products sold in the global market.

The global market for seafood alternatives is witnessing an exponential growth. Nevertheless, the nutritional quality of such products is scarcely studied. Thus, this study aimed to evaluate, for the first time, the nutritional quality of seafood alternatives launched in the global market from 2002 to 2021 and to compare them with the conventional seafood products. Using the Mintel Global New Products Database, the nutritional information of seafood alternatives (i.e., tuna, shrimps, calamari, fish fingers, fish sticks, salmon, caviar, and fillet) was retrieved, and compared with conventional products. A total of 149 seafood alternatives were identified, of which 83 items had complete mandatory nutritional labeling. Conventional products (n = 973) were also collected, from which 130 products have a complete nutritional labeling. Results revealed that tuna, shrimps, caviar and fillet alternatives contained significantly less protein than conventional products, while calamari, fish fingers, fish sticks and salmon alternatives had similar amounts to their conventional counterparts. Salt content was significantly higher in tuna, fish fingers and sticks substitutes, but lower in shrimps, calamari and caviar alternatives compared to conventional products. Overall, the commercially available seafood alternatives have nutritional strengths and some shortcomings to be further addressed in future research such as low protein content. Additionally, fortification of seafood alternatives with micronutrients, such as omega-3 fatty acids and vitamins (A, B, and D), should be considered to ensure a nutritional equivalence with the conventional products. Supplementary Information: The online version contains supplementary material available at 10.1007/s00217-022-04004-z.

Taurine, homotaurine, GABA and hydrophobic amino acids content influences "in vitro" antioxidant and SIRT1 modulation activities of enzymatic protein hydrolysates from algae.

Prevention and control of diseases and delaying the signs of ageing are nowadays one of the major goals of biomedicine. Sirtuins, a family of NAD+ dependent deacylase enzymes, could be pivotal targets of novel preventive and therapeutic strategies to achieve such aims. SIRT1 activating and inhibiting compounds, such as polyphenols and bioactive peptides, have been proposed to be involved in the development of many human diseases. The objective of this work was to assess and compare the antioxidant and SIRT1 modulation activities of enzymatic protein hydrolysates (EPHs) from a wide number of algae species (24 commercial samples and 12 samples harvested off the Atlantic coast of northern Spain). High antioxidant activities were observed in EPHs from red and green seaweed species. Moreover, 19 samples exhibited SIRT1 activation, while EPHs from the 16 samples were SIRT1 inhibitors. Pearson's correlation test and Principal Component Analysis revealed significant correlations between (1) total peptide and hydrophobic amino acid content in EPHs and their antioxidant activities, and (2) concentrations of taurine, homotaurine, and amino acid gamma aminobutyric acid in EPHs and their SIRT1 modulation activity.

Assessment of terahertz spectroscopy to detect antibiotic residues in food and feed matrices.

We report the use of terahertz (THz) spectroscopy to explore the spectral properties of eleven antibiotics commonly used in livestock production. Eight of the eleven antibiotics showed specific fingerprints in the frequency range between 0.1 and 2 THz. The main spectral features of two antibiotics (doxycycline and sulfapyridine) were still detectable when they were mixed with three food matrices (feed, milk, and egg powder). These preliminary results indicate that THz spectroscopy could be suitable for screening applications to detect the presence of antibiotic residues in the food industry, with the prospect to allow inspections directly on the production lines. THz spectroscopy is a non-destructive, non-contact, and real-time technique that requires very little sample preparation. Moreover, THz radiation can penetrate plastic and paper, which enables the detection of antibiotics in packaged food.

Analysis of eleven phenolic compounds including novel p-coumaroyl derivatives in lettuce (Lactuca sativa L.) by ultra-high-performance liquid chromatography with photodiode array and mass spectrometry detection.

INTRODUCTION: Lettuce is a widely consumed vegetable and a good source of phenolic compounds. Several factors (genetic, agronomical and environmental) can influence the lettuce composition; their effects are not completely defined and more studies are needed on this topic. OBJECTIVE: To develop an improved ultra-high-performance liquid chromatography (UHPLC) method to quantify the main target intact phenolic compounds in lettuce. METHODOLOGY: UHPLC identification of the compounds was supported by PAD spectra and MS(n) analyses. Quantification was carried out by PAD, by creating matrix-matched calibration curves at the specific wavelength for each compound. RESULTS: Sample pretreatment was simplified, with neither purification nor hydrolysis steps. Chromatographic conditions were chosen to minimise matrix interferences and to give a suitable separation of the major phenolic compounds within 27 min. The method allowed the quantification of 11 intact phenolic compounds in Romaine lettuces, including phenolic acids (caffeoyl and p-coumaroyl esters) and flavonoids (quercetin glycosides). Four p-coumaroyl esters were tentatively identified and quantified for the first time in lettuce. CONCLUSION: The main intact phenolic compounds, including four novel p-coumaroyl esters, were simultaneously quantified in lettuce with optimal performances and a reduced total time of analysis. These findings make headway in the understanding of the lettuce phytochemicals with potential nutritional relevance.

Food and Beverages Containing Algae and Derived Ingredients Launched in the Market from 2015 to 2019: A Front-of-Pack Labeling Perspective with a Special Focus on Spain.

Algae are a source of functional ingredients, with a large spectrum of healthy and functional compounds. Therefore, this study aimed to provide an overview on commercialized food and beverages made from algae and derived ingredients, with emphasis on the Spanish market, relying on the front-of-pack labeling. For this reason, the Mintel Global New Products Database was searched for foods and beverages containing "algae" ingredients, launched during the period 2015-2019. A total of 13,090 items were found worldwide, including 5720 items in Europe, in which 436 items were in Spain. Regardless of the market (global, European, and Spanish), a similar number of products categories (n = 20), dominant categories (dairy and desserts and ice cream) and dominant algal ingredient (carrageenans) were found. Nutritional information retrieved from Spanish products underlined that algae-based snacks had significantly lower energy, fat, and salt content compared to algae-free counterparts. On the contrary, spirulina- enriched ready to drink beverages had significantly higher energy and salt than algae-free. As such, reading the nutritional labeling is crucial to selecting products that suit consumer needs or/and expectations. Furthermore, only 8% of products reported the algal species and the level of inclusion, so this study emphasizes the importance of labeling legislation to provide complete product information to consumers.

Veggie burgers in the EU market: a nutritional challenge?

Nutritional information of burgers launched in the EU market during 2020 was retrieved from their labels. Products were initially classified into four types: i.e., veggie, red meat-, fish- and poultry-based. Gluten, wheat, and soy were the most declared allergens regardless of the burgers type. Veggie burgers showed levels of energy, fat, and saturate fatty acids (SFA) similar to fish- and poultry-based burgers, but lower than red meat burgers. Compared to conventional burgers, veggie had higher amounts of carbohydrates and sugars and a lower content of proteins, but no difference in salt. Due to the high compositional intra-variability in the veggie burgers, vegetarian and vegan burgers were further analyzed separately. Vegan burgers had higher levels of energy, fat, SFA and protein than vegetarian burgers, but lower carbohydrates and sugars. Once again large differences in the nutritional parameters were observed in both vegan and vegetarian burger categories due to the wide range of ingredients and formulations employed in these types of products. A clear and comprehensive informative labeling is especially needed for vegan and vegetarian burgers to allow the consumer to make a rational decision based on the nutritional facts of these products. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00217-021-03808-9.

Development of High-Protein Vegetable Creams by Using Single-Cell Ingredients from Some Microalgae Species.

The aim of this paper was to develop high-protein vegetable creams through the incorporation of microalgae. Single-cell ingredients from Arthrospiraplatensis (spirulina), Chlorella vulgaris, Tetraselmis chui, and Nannochloropsis oceanica were incorporated at two levels of addition (1.5% and 3.0%) to a standard vegetable cream (STD). Effects of incorporation were assessed in terms of physicochemical and rheological attributes as well as nutritional labeling facts. Creams formulated with 3% A. platensis, N. oceanica, or T. chui showed strong color differences (6 < ΔE < 12) compared to STD; creams formulated with 1.5% A. platensis, T. chui, or N. oceanica showed perceptible differences (3 < ΔE < 6); and those made with C. vulgaris at 1.5 and 3% exhibited small differences (ΔE < 2). Moisture content, water activity, pH, syneresis, and °Brix did not show significant changes. Adding microalgae increased Bostwick consistency and decreased the consistency coefficient (K) except in creams made with A. platensis, which showed comparable values to STD. Principal component analysis indicated that creams made with 1.5% C. vulgaris were the most similar to STD considering all evaluated parameters. Estimation of the nutritional labeling facts showed that the four formulations could be labeled as having "high protein content" following the present EU legislation.

Vegan Alternatives to Processed Cheese and Yogurt Launched in the European Market during 2020: A Nutritional Challenge?

Vegan alternatives to cheese (VAC) and yogurt (VAY) are fast-growing markets in Europe due to the increasing interest in plant-based alternatives to dairy products. This study aimed to take a closer look at the year 2020 and accordingly retrieved the nutritional information of dairy cheese and yogurt and their vegan counterparts for comparison. It was found that VAY (n = 182) provide more energy, total fats, and carbohydrates than dairy yogurt (n = 86), while saturated fatty acids (SFAs), sugars, and salt were not different between the two categories. Compared to dairy products (25.6%), 72.9% of the alternative products were declared low/no/reduced allergen, hence providing a larger spectrum of products to respond to consumers' requirements. VAC (n = 114) showed high versatility of form compared to dairy (n = 115). Nutritionally, VAC have higher total fats, SFAs, and carbohydrates, but lower protein, salt, and sugar than dairy cheese. Food developers will continue to look for clean label solutions to improve the nutritional values of vegan products through the incorporation of natural ingredients, besides enhancing their taste and texture to appeal to flexitarians.

Impact of Diets Including Agro-Industrial By-Products on the Fatty Acid and Sterol Profiles of Larvae Biomass from Ephestia kuehniella, Tenebrio molitor and Hermetia illucens.

Rearing insects on agro-industrial by-products is a sustainable strategy for the circular economy while producing valuable products for feed and foods. In this context, this study investigated the impact of larvae diet containing agro-industrial by-products on the contents of fatty acids and sterols of Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae), Tenebrio molitor (L.) (Coleoptera: Tenebrionidae), and Hermetia illucens (L.) (Diptera: Stratiomyidae). For each insect, selected diets were formulated using single or combined agro-industrial by-products (i.e., apricot, brewer's spent grain and yeast, and feed mill) and compared to a control diet. Fatty acid profiles showed differences depending on diet composition, but mostly depended on species: H. illucens was characterized by the abundance of C12:0, C16:0 and C18:2, whereas C:16, C18:1(n-9c), and C18:2(n-6c) were predominant in T. molitor and E. kuehniella. Sterols significantly varied as a function of diet composition and species. H. illucens showed low cholesterol levels and high campesterol and ß sitosterol levels (0.031, 0.554 and 1.035 mg/g, respectively), whereas T. molitor and E. kuehniella had high cholesterol and low campesterol contents (1.037 and 0.078 g/kg, respectively, for T. molitor; 0.873 and 0.132 g/kg, respectively, for E. kuehniella).