Alternative Protein Research Articles

A new protein source for pet food: cultivated meat

Cultivated meat is an alternative protein source developed to address the sustainability, public health and animal welfare concerns of conventional meat production. Hundreds of startups and academic institutions worldwide are working to make cultivated meat a cost-effective protein source for humans. However, cultivated meat could also be used to feed dogs and cats, contributing to solving the meat supply issues that the growing pet food market has been facing in recent years. The advantages of using cultivated meat as a protein source for pets would include a reduction of the environmental impact of pets' diets, decreased farm animal suffering and several benefits in the One Health framework, as cultivated meat-based pet food would significantly decrease the risk of spreading food safety pathogens, zoonotic diseases and resistant bacteria. The antibiotic-free manufacturing process and the aseptic conditions the cells require to grow in the bioreactors lead to these public health advantages. However, cultivated meat has never been produced at scale for human or pet consumption. Several technical challenges need to be overcome to make cultivated meat-based pet food prices accessible to consumers. As a novel ingredient, there is also no evidence of the effect of feeding cultivated meat to dogs and cats. In principle, cultivated meat can be both safe to be consumed long-term and nutritionally adequate – and with several possibilities for nutritional enhancement, potentially even superior to its conventional counterpart. However, the safety and nutritional soundness of cultivated meat-based products must be demonstrated by manufacturers to gain regulatory approval and favour consumer adoption. Veterinarians, veterinary nurses and technicians will play a critical role in the development of this new ingredient in many aspects, including product development, assessing safety and nutrition, conducting research and informing consumers. This review summarises the benefits and challenges of using cultivated meat as a pet food ingredient.

Environmental impacts of the filamentous fungi Paecilomyces variotii (PEKILO®) as a novel protein source in feeds for Atlantic salmon (Salmo salar)

The present study investigated environmental impacts associated with greenhouse gas (GHG) emission, economic Fish in Fish out ratio (eFIFO), waste outputs, and food-feed competition of feeding diets containing the filamentous fungus Paecilomyces variotii (PEKILO®) to Atlantic salmon (Salmo salar) pre-smolts. To achieve this, a feeding trial was conducted using four experimental diets. The control diet was based on fishmeal, soy protein concentrate, and wheat gluten meal. Diets 1, 2, and 3 were formulated so that P. variotii replaced 5, 10, and 20 % of the crude protein (CP) content of the control diet originating mainly from fishmeal, soy protein concentrate, and wheat gluten meal. Dietary inclusion of P. variotii affected simulated environmental indices such as GHG emission (P < 0.001); this index was P. variotii-dose-dependent, as illustrated by negative linear models (P < 0.001, adjusted R-squared = 0.96). Dietary inclusion of P. variotii was negatively correlated with eFIFO (P < 0.001, adjusted R-squared = 0.82). Dietary inclusion of P. variotii reduced food-competition feedstuff (FCF) in the diets (P < 0.001), as well as the amount of FCF to produce one kg of Atlantic salmon (P < 0.001). Relative to the control diet, solid nitrogen waste output, expressed as g kg−1 fish produced, was increased (P < 0.0001) in fish fed Diet 3. Conversely, linear decreases in solid phosphorous, magnesium and potassium (P < 0.0001) wastes expressed as g kg−1 fish produced were associated with increasing dietary inclusion of P. variotii. Likewise, linear decreases in dissolved nitrogen, magnesium, and potassium (P < 0.0001) wastes were observed with increasing replacement of the CP content of the control diet with P. variotii. Based on this assessment, P. variotii is a relevant alternative protein source with a low environmental footprint for use in Atlantic salmon feeds. Future novel feeds for Atlantic salmon should expand the use of alternative protein- and lipid-rich materials with a low ecological footprint that are produced according to a circular bioeconomy principle and utilize non-food-competing feedstuff (NFCF) such as P. variotii to move the industry toward sustainability.

Influence of pH and salt conditions on extraction efficiency and functional properties of Macrotermes nigeriensis protein concentrate for food applications

The search for alternative proteins from non-conventional sources for food processing has become important due to the shortage of conventional protein sources. Protein extractability from Macrotermes nigeriensis and its functional properties were investigated under various conditions of pH (2–10) and salt concentration (0.1–1.0 M). Extracted proteins were precipitated through pH adjustment and micellization, respectively. Results showed that maximum protein extractability was 68% and 62.1% at 0.5 M salt concentration and pH10.0, respectively. Salt-extracted protein-rich fraction (SP) had the highest protein composition (68.68 ± 0.41 g/100 g), seconded by alkaline-extracted protein-rich fraction (AP) (62.91 ± 0.53 g/100 g), compared with the raw and defatted fraction (34.36 ± 0.44 and 42.12 ± 0.15 g/100 g), respectively. The highest emulsion capacity (49%) and emulsion stability (35%) were recorded in alkaline pH 10.0. In comparison, the highest foaming capacity (24%) and foaming stability (16%) were recorded in the salt-extracted fraction at 6%. This information could be useful for further studies on the food application potential of proteins isolated from edible M. nigeriensis.Graphical

Effect on Growth Performance and Nutritive Value of Cultivated Azolla filiculoides As An Alternative Feedstuff for Ruminant.

Azolla filiculoides is a tiny, free-floating aquatic fern and has a potential alternative protein and fibre source for ruminants, was investigated for its cultivation optimisation and feedstuff suitability. Study 1 was conducted to investigate the influence of different fertiliser types (control, broiler manure, sheep manure, cow manure) and concentrations (0.25 g/L-1.25 g/L) on the growth performance (fresh weight, doubling time, relative growth rate) and nutrient composition (dry matter, ash, crude protein, crude fibre, crude fat) of A. filiculoides. The optimised type of fertiliser and concentration in Study 1 were further adopted in Study 2 to evaluate the effect of different fertiliser processing methods on the growth performance, nutritive value and in vitro rumen digestibility of A. filiculoides upon cultivation. The findings in Study 1 showed that cultivation of A. filiculoides using sheep manure at the concentration of 1.00 g/L is the best resulted in the shortest doubling time (3 to 5 days) and produced fresh weight (FW), relative growth rate (RGR), crude protein (CP) and crude fibre (CF) at 132.2 g/m2, 0.32 g/g/day, 21.2% DM-1 and 14.4% DM-1, respectively. Furthermore, unprocessed sheep manure (T3) exhibited superior (p < 0.05) fresh weight, relative growth rate, nutrient composition and fibre components compared to the burned manure treatment (T2). In vitro digestibility analysis discovered that T3 achieved a 24-hour accumulated gas production of 86.9 mL DM-1, with in vitro dry matter digestibility (IVDMD), in vitro organic matter digestibility (IVOMD) and metabolisable energy (ME) of 82.9%, 43.7% and 5.8 MJ/kg DM, respectively. These findings suggest that Azolla filiculoides cultivation can be economically optimised using 1.00 g/L unprocessed sheep manure (fresh manure), potentially serving as a self-produced, nutritious feedstuff for ruminants.

Sago palm weevil larvae: larval ontogeny and nutritional value assessed from enzymatic activity and in vitro digestibility

Abstract Sago palm weevil larvae (SPWL, Rhynchophorus vulneratus) have potential for use as food and feed. This study aimed to investigate their development, digestive enzyme activity, nutrient values, and digestible protein using an in vitro digestibility technique. Bioassays for SPWL at 21, 28, 35, 42, 49, and 56 days after hatching (DAH) were conducted in a completely randomized design (CRD). Analysis of variance (ANOVA) was performed, and means were compared at using Bonferroni (Dunn) t-tests and orthogonal polynomial contrasts. Trypsin activity significantly increased at 35 DAH (, quadratic effect ), with peak chymotrypsin activity at 28 DAH (). Lipase activity showed both linear and quadratic increases (), and cellulase activity also increased significantly (, linear and quadratic terms ). Meanwhile, the A:T ratio increased significantly (, quadratic effect ) with age-related rises in these digestive enzyme activities. The highest crude protein content in SPWL was observed at 42 DAH (), with peak carbohydrate at 28 DAH (). Phenylalanine and histidine levels increased linearly (), while lysine and total essential amino acids peaked at 42 DAH (, linear and quadratic terms ). Levels of linoleic acid, total polyunsaturated fatty acids, and the n-6:n-3 ratio peaked at 49 DAH (). In vitro protein digestibility assays were performed using enzymes extracted from quails, broilers, Thai native chickens, and ducks. Significant increases in digestibility were attributed to SPWL protein content and age, particularly beyond 42 DAH (, linear and quadratic terms ), except with duck enzymes. In conclusion, these findings suggest that SPWL cultivation, particularly between 42 and 56 DAH, provides a potential alternative protein source with substantial nutritional benefits for poultry feed. The promise of SPWL to contribute to more sustainable and efficient poultry feeding practices is evident, and further in vivo studies are warranted to validate these results.

Alternative protein sources in the European diets – integrating health risk‐benefit and sustainability ‘ALTERNATIVA’

Abstract The pressure global food systems exert on the environment has amplified over time, following the growth of the human population, changes in the food consumption patterns and intensification of the production systems. Without changes in food systems, an overpressure on agriculture and food production will be unavoidable, with inevitable consequences for the environment and human health. Animal‐derived protein production like meat and milk is among the most important drivers of this impact. Under this context, the evaluation of the health and sustainability impacts of alternatives for animal‐based food proteins are central aspects of this discussion. Health risk‐benefit assessment (RBA) of foods estimates the overall human health impact following exposure to a particular food or food component, integrating risks and benefits in comparable measures. Methods that simultaneously consider the environmental impact of the food production systems were needed to perform a sustainable holistic assessment. “ALTERNATIVA | Alternative protein sources in the European diets – integrating health risk‐benefit and sustainability” aimed to develop a holistic approach to assess the impact of alternative protein sources, integrating health and sustainability assessment. This project, considering five complementary work packages, developed activities to enhance the capacity‐building among members of the consortium in applying methodologies related to the health RBA and sustainability impact assessment; to develop, in collaboration with different stakeholders, a protocol to integrate the assessment of the impact of foods/diets on health and on sustainability; to apply the derived framework to a case study, to validate it with a real case scenario, estimating the health and sustainability impacts of replacing red meat consumption by pulses in two different countries; and to disseminate and promote the established method of integrated health and sustainability assessment to other potential EU users. This external scientific report summarizes the main outputs and results obtained in the context of the ALTERNATIVA project.

Silk worm pupal meal: A alternative protein source in aqua feed

Silk worm pupal meal: A alternative protein source in aqua feed

High dietary Mucuna pruriens utilis seed meal compromises growth performance, carcass traits, haemato-biochemistry, and meat quality of broilers.

Usage of soyabean meal (SBM) in broiler diets is economically and environmentally unsustainable thus necessitating investigation of alternative protein sources. Therefore, this study investigated effects of incremental inclusion levels of Mucuna pruriens utilis seed meal (MSM) for partial substitution of SBM in broiler diets. In a completely randomized design (CRD), 400day-old Ross 308 chicks were allotted to 5 iso-caloric-nitrogenous MSM-containing (0, 5, 10, 15, and 20%) dietary treatments. Each treatment was replicated 8 times, with each pen having 10 birds, during starter (d1 - 14), grower (d15 - 28), and finisher (d29 - 42) phases. Results showed that dietary MSM decreased feed intake (FI: quadratic: P < 0.001), body weight gain (BWG: linear: P < 0.001), and feed conversion efficiency (FCE: linear: P < 0.001) as it linearly decreased slaughter weight (SW: P < 0.001), hot carcass weight (HCW: P < 0.001), cold carcass weight (CCW: P < 0.001), dressing percentage (P < 0.001), and breast weight (P < 0.05). In contrast, dietary MSM linearly increased the weights of the liver (P < 0.01), proventriculus (P < 0.001), gizzard (P < 0.001), duodenum (P = 0.01), jejunum (P < 0.001), ileum (P < 0.001), caecum (P < 0.01), and colon (P < 0.01). Also, dietary MSM quadratically increased blood heterophils (P < 0.05) and alkaline phosphatase activity (P < 0.05) of the chickens whilst linearly increasing their serum amylase (P = 0.001) and lipase (P = 0.001) activities and linearly decreasing their serum symmetric dimethylarginine (SDMA: P = 0.001) and cholesterol (P < 0.05). Further, dietary MSM linearly decreased chicken breast meat ultimate pH (P < 0.05) whilst linearly increasing its cooking loss (P < 0.01), drip loss (P < 0.05) and shear force (P < 0.01). In conclusion, dietary MSM compromised growth performance, carcass characteristics, and meat quality of broilers as it increased the weights of their digestive-metabolic organs.

Knowledge of and Willingness to Consume Insects as an Alternative Protein Source Among College Students in the Midwest

Knowledge of and Willingness to Consume Insects as an Alternative Protein Source Among College Students in the Midwest

A study on safety evaluation system of cultured foods among alternative proteins

AbstractFood produced by cell culture has been approved in Singapore and the United States. Food safety evaluation systems of Singapore, the United States, Europe, and Korea were reviewed for producing cultured foods with new technologies. In Singapore, Europe, and Korea, safety evaluation of cultured meat is conducted by applying for such evaluation in the Novel Food system. In contrast, in the case of the United States, safety evaluation for cultured meat is conducted by the FDA and the USDA, considering it as an altered product of production methods, not a novel food. Required safety assessment data vary depending on whether the cultured meat is a novel food or a different food with a different production method. Accordingly, the current study presents differences between required documents according to the two distinct perspectives.

Lactational performance of dairy buffaloes affected by replacing soybean meal with an alternative microbial protein source

Thirty dairy buffaloes were used for 90 days to evaluate the impact of a novel protein source (HI-PRO®), a feed alternative rich in protein (made from Saccharomyces and Bacillus), on the productivity of nursing buffaloes. The nursing water buffaloes had an average weight of 550 ± 11.2 kg. The treatments consisted of diets containing two distinct protein sources: (1) soybean meal (44 % as the control group); and (2) HI-PRO® product. Throughout the trial, measurements of nutritional digestibility and blood metabolites (total protein, albumin, urea, and creatinine) were conducted. Quantity and composition of the milk were measured to determine the content of milk proteins, lipids, and lactose. The outcome of the results showed insignificant decrease in the buffaloes receiving HI-PRO® feed versus control group. Furthermore, there was enhancement in the fiber digestibility by approximately 2.5 % in the HI-PRO® group compared to the control group. The levels of protein, albumin, globulin, urea, and creatinine fall within the normal range for animals in good health. Using HI-PRO® resulted in a marginal improvement in milk production, increasing it by approximately 4.8 % compared to soybean meal. Furthermore, the buffaloes fed HI-PRO® showed a slightly elevated 4 % fat-corrected milk output and milk composition. To summarize, nursing buffaloes can utilize HI-PRO® as an efficient protein source in their diets, replacing soybean meal.

Deciphering the ghost proteome in ovarian cancer cells by deep proteogenomic characterization

Proteogenomics is becoming a powerful tool in personalized medicine by linking genomics, transcriptomics and mass spectrometry (MS)-based proteomics. Due to increasing evidence of alternative open reading frame-encoded proteins (AltProts), proteogenomics has a high potential to unravel the characteristics, variants, expression levels of the alternative proteome, in addition to already annotated proteins (RefProts). To obtain a broader view of the proteome of ovarian cancer cells compared to ovarian epithelial cells, cell-specific total RNA-sequencing profiles and customized protein databases were generated. In total, 128 RefProts and 30 AltProts were identified exclusively in SKOV-3 and PEO-4 cells. Among them, an AltProt variant of IP_715944, translated from DHX8, was found mutated (p.Leu44Pro). We show high variation in protein expression levels of RefProts and AltProts in different subcellular compartments. The presence of 117 RefProt and two AltProt variants was described, along with their possible implications in the different physiological/pathological characteristics. To identify the possible involvement of AltProts in cellular processes, cross-linking-MS (XL-MS) was performed in each cell line to identify AltProt-RefProt interactions. This approach revealed an interaction between POLD3 and the AltProt IP_183088, which after molecular docking, was placed between POLD3-POLD2 binding sites, highlighting its possibility of the involvement in DNA replication and repair.

Study on Rumen Degradability and Intestinal Digestibility of Mutton Sheep Diets with Different Concentrate-to-Forage Ratios and Nonfiber Carbohydrates/Neutral Detergent Fiber Ratios.

This study was conducted to investigate the rumen degradability and intestinal digestibility of mutton sheep diets different in concentrate-to-forage ratio, NFC/NDF, and ingredient combination, providing a guideline for the selection of a fattening diet for mutton sheep. Twenty-eight diets composed of four raw material combinations and seven concentrate-to-forage ratios and four three-year-old mutton sheep with permanent rumen fistulas were used in the experiments. The nutrient composition of the diets was first analyzed, and then an in situ method and in vitro three-step method were separately used to measure the rumen degradability and intestinal digestibility, mainly focusing on the effects of dietary concentrate-to-forage ratio and NFC/NDF as well as the effects of soybean meal and soybean meal replacement and peanut vine and peanut vine replacement. The results showed that a dietary concentrate-to-forage ratio of 70:30~80:20 and an NFC/NDF ratio of 1.5~2.0 are recommended for fattening mutton sheep, and low-cost cottonseed meal and rapeseed meal can be feasible alternative protein sources to soybean meal. In addition, the nutritional values of sunflower seed hulls and rice hulls for mutton sheep are lower than that of peanut vine. Such a study can provide practical guidelines for enterprises and farmers, being of important significance for the high-quality development of the mutton sheep industry.

Fermented but Not Irradiated Cottonseed Meal Has the Potential to Partially Substitute Soybean Meal in Broiler Chickens.

This study was conducted to investigate and compare the effects of substituting soybean meal (SBM) with untreated cottonseed meal (CSM), fermented CSM (FCSM), or electron beam-irradiated CSM (ICSM) on the growth performance, cecal microbiota, digestive enzyme activity, apparent ileal digestibility (AID), and excreta gas emission of broiler chickens. A total of 384 one-day-old male broiler chickens were randomly assigned to four experimental diets, with eight replicates per diet and 12 birds per replicate, for six weeks. The experimental diets consisted of a control diet based on corn-SBM and three other diets in which 50% of the SBM (control) was substituted with CSM in its raw, irradiated, and fermented forms. The results showed that throughout the entire rearing period, feeding broiler chickens with ICSM significantly increased average daily gain (ADG) and body weight (BW) compared to the CSM diet (p < 0.05). Replacing 50% of SBM with FCSM led to a significant improvement in BW, ADG, and feed conversion ratio (FCR) compared to the CSM and ICSM diets (p < 0.05). Interestingly, no significant differences in BW, ADG, or FCR were observed between birds fed FCSM and those on the control diet (p > 0.05). Birds fed FCSM diets exhibited the lowest pH value in the crop, ileum, and ceca. Substituting SBM with FCSM significantly reduced Escherichia coli and Clostridium spp. counts in the ceca, while enhancing the presence of Lactobacillus spp. (p < 0.05). The AID of protein and ether extract was higher in the FCSM group than in the CSM and ICSM groups (p < 0.05). Compared to the CSM diet, ICSM feeding improved protein digestibility (p < 0.05). Broiler chickens on the FCSM diet exhibited higher intestinal amylase and protease activity than those on the other diets (p < 0.05). Furthermore, feeding diets containing FCSM significantly reduced ammonia emissions compared to the other diets (p < 0.05). Overall, our results indicated that microbial fermentation of CSM is a more effective approach than irradiation for enhancing the nutritional value of CSM. Therefore, FCSM is recommended as a viable alternative protein source that can safely replace up to 50% of SBM in broiler chicken diets, particularly during times of fluctuating SBM prices and availability issues.

Additive effects of Bridelia ferruginea leaf meal as a partial replacement for soybean meal on nutrients digestibility, growth performance, and carcass characteristics of weaner rabbits

BackgroundProtein is a crucial nutrient for enhancing livestock performance and productivity, traditionally sourced from soybean and fish meal. However, issues like erratic availability and rising prices necessitate exploring alternative protein sources. Tropical plants, especially leaf meals, are being considered to replace or supplement conventional protein sources in rabbit diets. Bridelia ferruginea, a plant abundant in West Africa, is recognized for its medicinal properties, but its potential as an animal feed is largely unexplored. This study aims to evaluate the impact of graded levels of Bridelia ferruginea leaf meal (BFLM) as a partial substitute for soybean meal in the diets of weaner rabbits.Materials and methodsThe experiment was conducted at the Animal Science Department of Kwame Nkrumah University of Science and Technology in Ghana. BFLM was obtained by air-drying harvested leaves and milling them. Four diets were formulated, with BFLM partially replacing soybean meal at levels of 0%, 10%, 20%, and 30%. Representative samples of BFLM and all formulated diets were analyzed for their proximate composition and detergent fibre fractions. Twenty-four weaner rabbits were assigned to these diets in a Completely Randomized Design. The study covered a 10-week period, assessing growth performance, nutrient digestibility, carcass characteristics, and economics of production.ResultsThe chemical composition of BFLM revealed favorable attributes for rabbit feeding, with 20.5% crude protein and 92.6% dry matter. Growth performance declined at 30% BFLM inclusion, indicating a possible impact of anti-nutritional factors. Feed conversion ratio increased at higher BFLM levels. Nutrient digestibility showed a decline in dry matter digestibility at 30% BFLM. Carcass characteristics were affected at higher BFLM levels, with dressing percentage decreasing. Weights of internal organs were generally not negatively affected by the inclusion of BFLM. Economic analysis indicated a linear reduction in feed cost with increasing BFLM inclusion.ConclusionThe study suggests that BFLM can be incorporated up to 20% in weaner rabbit diets without compromising productivity, nutrient digestibility, or dressing percentage. Beyond 20%, adverse effects on growth performance and some carcass characteristics were observed. Economically, BFLM incorporation up to 20% lowered feed costs without compromising feed efficiency. Incorporation of BFLM up to 20% in weaner rabbit diets had no adverse effects on the weights and functions of the heart, liver, and kidneys. Overall, BFLM presents a promising alternative protein source for weaner rabbit diets, contributing to cost-effective and sustainable livestock production.

Blood Parameter Response in Growing Alpine Goat Kids Fed Diets Containing Extruded Flaxseed or Pumpkin Seed Cake

Blood parameters can provide information on the nutritional status of goat kids, which is related to both health and performance. The present study aimed to research whether feeding extruded flaxseed (FS) and pumpkin seed cake (PC), as an alternative protein source in diets, has an effect on the hematological and serum biochemical parameters of goat kids during growth. In the small-scale goat farm, 31 French Alpine goat kids aged 32 days were used for the study. The goat kids were subjected to three different feeding treatments: a mixture containing soybean meal and extruded soybeans (CON), a mixture containing 16% PC (PC-16), and a mixture containing 9% FS (FS-9). They were monitored during the suckling, weaning, and post-weaning growth periods. PC-16 and FS-9 in goat kids’ diets did not result in any changes regarding average daily weight gain. The WBC count was higher in goat kids fed FS-9 and PC-16 compared to CON (9.84 and 9.54 vs. 6.61 × 109 L) diets during the weaning period. GGT activity was lowest in the serum of goat kids fed PC-16 compared to CON post-weaning (38.65 vs. 48.40 U/L). In addition, FS-19 increased GPx compared to kids fed PC-16 post-weaning (809.7 vs. 600.8 U/L). Regarding blood parameters, PC-16 and FS-9 can be used in goat kids’ nutrition as alternative sources of proteins on a small-scale goat farm without compromising goat kids’ growth.

Multifunctional properties of peptides derived from black cricket (Gryllus assimilis) and effects of in vitro digestion simulation on their bioactivities

Insects are a rich source of proteins and are produced in systems that have lower environmental impact. As an alternative protein source, they can be consumed directly or used as an ingredient in other formulations. Recently, there has been growing interest in utilizing insect proteins as a substrate to obtain bioactive peptides as well as in investigating the maintenance of their biological properties under physiological conditions. This study aimed to evaluate the impact of simulated digestion on the bioactive properties of protein hydrolysates from black crickets (Gryllus assimilis). Following simulated digestion of the hydrolysate obtained through the application of Flavourzyme, the scavenging activities of ABTS and DPPH radicals, and ferric reducing antioxidant power (FRAP) increased by approximately 17 %, 246 %, and 173 %, respectively. For the hydrolysate obtained using the binary combination of Flavourzyme/Neutrase, the inhibitory activities of α-amylase and α-glucosidase after digestion were 47.87 % and 12.73 %, respectively, not significantly (p > 0.05) different from non-digested hydrolysates. The angiotensin-converting enzyme (ACE) inhibitory activity of the sample hydrolyzed with Flavourzyme/Alcalase proteases was 42.22 %, but this property was completely lost after in vitro digestion. Untargeted proteomic analysis allowed the identification of 22 peptides in the <3 kDa fraction of the digested black cricket protein. The LPPLP sequence was considered potentially bioactive for all activities tested in silico.

Comparative efficacy of amino acid availability and peptidomic analysis of alternative proteins from different sources under dynamic in vitro protein digestion

The increasing global demand for animal protein highlights the critical necessity of exploring alternative protein sources. Accessing the nutritional characteristics of emerging alternative proteins is vital for food processing and consumer acceptance. This study investigated the amino acid availability and peptidomics profiles of plant- (soy), fungal- (filamentous fungus Fusarium venenatum), microalgae- (Nannochloropsis oculata), and insect-based (black soldier fly) proteins using an advanced dynamic in vitro digestion system, with whey protein as a reference. After gastric digestion, soy and mycelial proteins, despite displaying distinct microstructures, exhibited slightly higher protein digestibility levels of 28.79% and 26.36%, respectively, compared to whey protein (22.39%). However, following small intestinal digestion, whey protein (61.31%) significantly outperformed alternative proteins. Mycelial protein (43.71%) exhibited digestibility similar to soy protein (46.12%), markedly surpassing both microalgae protein (33.35%) and insect protein (26.58%). Notably, insect protein achieved an amino acid availability nearly approaching that of whey protein, underscoring its substantial proteolysis capability. Furthermore, microalgae protein produced the highest percentage and number of bioactive peptides under simulated digestion, suggesting potential health benefits such as angiotensin I-converting enzyme (ACE) and dipeptidyl peptidase-IV (DPP-IV) inhibitor activities, which could mitigate the risk of hypertension and Type 2 diabetes. Alternative proteins from different sources demonstrate diverse digestive mechanisms and proteolysis profiles, thereby bolstering their potential applications as nutritious food ingredients in the future.

Predicting Mutation-Induced Allosteric Changes in Structures and Conformational Ensembles of the ABL Kinase Using AlphaFold2 Adaptations with Alanine Sequence Scanning.

Despite the success of AlphaFold2 approaches in predicting single protein structures, these methods showed intrinsic limitations in predicting multiple functional conformations of allosteric proteins and have been challenged to accurately capture the effects of single point mutations that induced significant structural changes. We examined several implementations of AlphaFold2 methods to predict conformational ensembles for state-switching mutants of the ABL kinase. The results revealed that a combination of randomized alanine sequence masking with shallow multiple sequence alignment subsampling can significantly expand the conformational diversity of the predicted structural ensembles and capture shifts in populations of the active and inactive ABL states. Consistent with the NMR experiments, the predicted conformational ensembles for M309L/L320I and M309L/H415P ABL mutants that perturb the regulatory spine networks featured the increased population of the fully closed inactive state. The proposed adaptation of AlphaFold can reproduce the experimentally observed mutation-induced redistributions in the relative populations of the active and inactive ABL states and capture the effects of regulatory mutations on allosteric structural rearrangements of the kinase domain. The ensemble-based network analysis complemented AlphaFold predictions by revealing allosteric hotspots that correspond to state-switching mutational sites which may explain the global effect of regulatory mutations on structural changes between the ABL states. This study suggested that attention-based learning of long-range dependencies between sequence positions in homologous folds and deciphering patterns of allosteric interactions may further augment the predictive abilities of AlphaFold methods for modeling of alternative protein sates, conformational ensembles and mutation-induced structural transformations.

Industrial processing of chickpeas (Cicer arietinum) for protein production

AbstractThe increasing global interest in plant‐based proteins stems from concerns about the environmental impact, sustainability, animal welfare, and health implications associated with consuming animal‐based proteins. In the frame of alternative protein sources, chickpea (Cicer arietinum) emerged as a rich source of dietary proteins besides containing good amount of carbohydrate, fat, and fiber. As a protein ingredient, chickpea is available in three forms, namely, flour, concentrate, and isolate. This chickpea protein can be extracted using both wet and dry fractionation methods where the former one includes wet extraction followed by isoelectric precipitation, while the later one indicates dry milling followed by air classification. However, different nonthermal emerging technologies have been seen to assist in extracting protein as well as modifying their functionalities. This review gives an outline of the recently available literature on composition, industrial processing and associated technological challenges, functionality, and application of chickpea protein ingredients. Furthermore, discussion on the modification/improvement of chickpea protein functionality with the assistance of emerging technologies and the potentiality of by‐products produced during chickpea protein processing are also included. Based on the available findings and discussion, it is seen that apart from being a comparable source of alternative animal‐based protein to extract, chickpea derived by‐products can also be a potential source of valued ingredients that might contribute to the circular economy.