Edible Insect Protein Research Articles

Edible insect protein concentrates: Optimized salt-assisted extraction methods evaluation

This study explores the extraction of proteins from edible insects such as Tenebrio molitor, Acheta domesticus, and Locusta migratoria using alkaline extraction and acid precipitation (AEAP) as a control method and evaluates the influence of salting-in (NaCl-assisted), salting-out ((NH₄)₂SO₄), and combined salting-in/out techniques on the resulting protein quality and functional properties. We hypothesized that salt-assisted methods would enhance protein extraction efficiency and functionality compared to AEAP. Molecular size distribution analysis confirmed that the salting-out method preferentially aggregated smaller proteins (<30 kDa). FT-IR spectroscopy revealed notable changes in protein secondary structure across extraction methods, while amino acid profiling identified 17 amino acids, with essential amino acids (EAAs) comprising 37.8–44.2 % of total amino acids. Salt-assisted methods significantly increased the zeta potential (up to −62.67 mV) and modulated particle size distribution (180–492 nm) compared to the control samples. Rheological properties varied with extraction techniques, with potential fluid-type transitions. Proteins extracted via salt-assisted methods demonstrated high purity (>70 %), enhanced solubility (>60 % at pH 7.4), improved oil- and water-holding capacities (1.40–8.09 g/g, 2.41–4.4 g/g), and superior emulsifying properties (EAI >47 m2 g−1, ESI >50 %). These findings highlight that salt-assisted extraction methods can improve the quality and functionality of insect protein concentrates, supporting their potential for food-grade applications.

Consumer sensory profiling and liking of Bolognese-type sauces: how do insect and plant foods really fare against red meat?

Abstract Meeting global targets for healthier and more sustainable diets calls for a substantial reduction of meat consumption in Western nations, especially red and processed meat. This requires a transition to the large-scale production, marketing, and adoption of alternative proteins. The current state of development of new plant- and insect-based foods holds good promise, but optimizing their sensory quality to the point where they can satisfactorily replace everyday meat-eating experiences remains a challenge, demanding a more consumer-oriented approach. This study investigated how Portuguese adults (, 18-40 years old, 50% female, regular meat eaters) perceived the sensory characteristics of seven Bolognese-style pasta sauces – made with 100% red meat (beef and pork), plant (soya beans or peas) or edible insect (house cricket or yellow mealworm larvae) protein – and how this affected their hedonic preferences for these products. Expectedly, the meat-based sauce was generally the most appreciated. Still, consumers liked all products slightly to moderately apart from mealworm mince. Sensory-mediated disgust affected the evaluations of some insect- and plant-based sauces, while others benefitted from formulations that enhanced savouriness and the prototypical attributes of a tomato pasta sauce. Moreover, the disconfirmation of positive sensory-affective judgements of ‘meatiness’ induced by mince-like texture cues penalized liking. High inter-individual variation in evaluations was observed. Distinct preferences for spiciness, for instance, moderated the effects of meat substitution on product liking, with over half of participants appreciating a mild or a spicy plant-based option more than the typical beef Bolognese. Sociodemographics, Beef Consumption, Healthy Eating Consciousness and Food Disgust Sensitivity were all important predictors of patterns of product liking, pointing out that the type of protein selected is just one of the many factors driving the acceptance of meat substitutes by consumers that can be leveraged by the food industry.

The Effect of Ultrasound Treatment on the Structural and Functional Properties of Tenebrio molitor Myofibrillar Protein.

The objective of this study was to explore the impacts of various ultrasonic powers (0, 300, 500, 700, and 900 W) on the structure and functional attributes of the myofibrillar protein (MP) of Tenebrio molitor. As the ultrasonic intensity escalated, the extraction efficiency and yield of the MP rose, while the particle size and turbidity decreased correspondingly. The reduction in sulfhydryl group content and the increase in carbonyl group content both suggested that ultrasonic treatment promoted the oxidation of the MP to a certain extent, which was conducive to the formation of a denser and more stable gel network structure. This was also affirmed by SEM images. Additionally, the findings of intrinsic fluorescence and FTIR indicated that high-intensity ultrasound significantly altered the secondary structure of the protein. The unfolding of the MP exposed more amino acid residues, the α-helix decreased, and the β-helix improved, thereby resulting in a looser and more flexible conformation. Along with the structural alteration, the surface hydrophobicity and emulsification properties were also significantly enhanced. Besides that, SDS-PAGE demonstrated that the MP of T. molitor was primarily composed of myosin heavy chain (MHC), actin, myosin light chain (MLC), paramyosin, and tropomyosin. The aforementioned results confirmed that ultrasonic treatment could, to a certain extent, enhance the structure and function of mealworm MP, thereby providing a theoretical reference for the utilization of edible insect proteins in the future, deep-processing proteins produced by T. molitor, and the development of new technologies.

Anti-inflammatory and anti-hyperglycemia effects of mealworm (Tenebrio molitor larvae) protein extracted by four methods: alkali, salt, enzyme, and screw press.

The use of edible insect protein in food products is contingent on their biological effects. Conventional protein extraction methods are not only time-consuming and costly but also energy-intensive. There is a need for alternative techniques that maintain the bioactivities of insect proteins and are environmentally sustainable. This study compares the health functionality of mealworm (Tenebrio molitor larvae) concentrates obtained by conventional methods-alkali and salt (MS) extraction-and nonconventional methods-enzyme (ME) and screw press (MP)-to enhance their applicability despite lower protein concentration. Overall, MP exhibited the highest essential amino acids content, whereas ME showed the highest in vitro digestibility, total phenolic contents, and antioxidant capacities among all the concentrates. ME also had a significant cell proliferative capacity at concentrations ≥500µg/mL. MS significantly inhibited tumor necrosis factor-alpha and interleukin-1beta secretion in lipopolysaccharide-treated Hep3B cells compared to other samples. As for anti-hyperglycemia effects, treatment with MS and ME for 2 and 5min significantly increased the p-Akt/Akt ratio (MS, 1.34- and 1.61-fold; ME, 2.26- and 2.70-fold, respectively). In conclusion, enzyme treatment enhanced nutritional value and antioxidant capacity, whereas salt treatment potentially contributed to anti-inflammatory and anti-hyperglycemia activities. Hybrid extraction techniques combining conventional and nonconventional methods are suggested based on target applications, considering health benefits, environmental impact, costs, and efficiencies. PRACTICAL APPLICATION: Four mealworm protein extraction methods (alkali/salt/enzyme/screw press) were compared for their nutritional and biological properties. Alkali extraction enhanced protein content, enzyme treatment improved nutritional value and antioxidant capacity, and salt-assisted extraction exhibited immunomodulatory effects in vitro. Notably, enzyme and salt treatments produced protein concentrates with significant antidiabetic and anti-hyperglycemic properties.

The Allergen Profile of Two Edible Insect Species-Acheta domesticus and Hermetia illucens.

Edible insect proteins are increasingly introduced as an alternative sustainable food source to address the world's need to feed the growing population. Tropomyosin is the main insect allergen; however, additional potential allergens are not well characterized and the impact of extraction procedures on immunological reactivity is unknown. Proteins from different commercial food products derived from cricket (Acheta domesticus) and black soldier fly (BSF) (Hermetia illucens) are extracted using five different extraction buffers. The proteins are analyzed by SDS-PAGE and immunoblotting using allergen-specific antibodies and crustacean allergic patient sera. IgE binding bands are analyzed by mass spectrometry as well as the complete allergen profile of all 30 extracts. Urea-based buffers are most efficient in extracting insect allergens. Shrimp-specific antibody cross-reactivity to tropomyosin from cricket and BSF indicates high sequence and structural similarity between shrimp and insects. Additional unique allergens are identified in both species, including hemocyanin, vitellogenin, HSP20, apolipophorin-III, and chitin-binding protein. Identifying potential allergenic proteins and their isoforms in cricket and BSF requires specific extraction approaches using urea-based methods. While tropomyosin is the most abundant and immunoreactive allergen, seven unique allergens are identified, highlighting the need for insect species-specific allergen detection in food products.

Cooking resistant edible crickets-specific peptides for authenticity testing of meat products

Abstract As consumer and manufacturer interests in edible insects and processed food with added insects are increasing, new possibilities for detecting edible insect proteins in processed foods have become increasingly important. In the present study, a proteomic strategy was applied to identify insect proteins and thermostable house cricket-specific (Acheta domesticus) peptide markers. To determine the limit of detection (LOD) for house cricket proteins, cooked meatballs containing house cricket protein powder (CP) as a partial pork substitute were investigated. The final concentration of CP ranged from 0.8% to 7.6%. The LODs for tropomyosin 1 and translational elongation factor-2 were 0.8% (w/w), whereas for apolipophorin-III it was 2.5% (w/w). Eight heat-resistant peptides unique to the family Grillidae (true crickets) and four peptides unique to the Acheta domesticus were identified. The results suggest that selected cricket-specific and processing-resistant peptide markers have potential utility in the authentication of the cricket formulations used in meat products. However, this has to be confirmed on different heavily processed meat products.

A review of edible insect proteins application: health benefits and safety

Abstract In recent years, the rapid growth of the global population has led to a serious shortage of food resources, necessitating the exploration and utilization of new resources. Edible insects have gained attention due to their high protein content, similar to that of traditional meat proteins, along with a host of beneficial nutritional and health properties. Besides their wonderful nutritional composition, edible insect proteins also have various health benefits like antioxidation, anti-hypertension, anti-inflammation, antimicrobial activity, and immunoregulation. However, considerations regarding their safety and hygienic characteristics are crucial for their widespread development as a new protein source. Herein, we systematically summarized the nutritional value and functional features of insect protein, additionally, their applications and health benefits were also reviewed. Finally, their safety aspects and prospect of further research were also discussed in order to provide beneficial references for future research.

A common edible insect (Antheraea assamensis) protein hydrolysate regulates LPS-induced oxidative stress and inflammation by modulating the TLR4/NF- κ β Signaling Pathway

Abstract Dietary intake of antioxidant and anti-inflammatory-rich foods is a growing approach for fighting inflammation and its associated disorders. Edible insects are gaining popularity as a food source; however, their therapeutic properties, anti-inflammatory activity and their mechanism of action remain largely unexplored. Herein, we evaluate the effectiveness of Antheraea assamensis pupae hydrolysates against oxidative stress and inflammation. The proteins isolated from the pupae were digested with pepsin or pancreatin, or pepsin + pancreatin and passed through a 30 kDa molecular cut-off membrane. The low molecular weight protein hydrolysates were characterised using RP-HPLC, HRMS, XRD, and FTIR-ATR. Hydrolysate from pepsin + pancreatin digestion showed significantly higher DPPH (77.13±2.57%), superoxide (72.08±1.15%) and hydroxyl (56.32±0.90%) radical scavenging activity in a cell-free system. Further, it reduced intercellular ROS production and suppressed the lipopolysaccharide-induced protein expression of TLR4, p-IKKβ, p-NF-, IL-1β, and COX2 in RAW 264.7 macrophages. Furthermore, it inhibited the secretion of pro-inflammatory mediators, including IL-6, IL-1β, and MIP-2. These findings suggest that A. assamensis pupae hydrolysates can be a potential source of bioactive peptides for managing oxidative stress and inflammation.

Immunomodulatory effect of earthworm protein autolysates on Cyclophosphamide(CTX)-Induced immunosuppressed mice

Earthworms are rich in protein and are considered a good source of edible insect protein. However, there has been limited research on the bioactivity of earthworm protein hydrolysates. The present study investigated the immunological impact of earthworm protein autolysate (EPA) on Cyclophosphamide (CTX)-induced immunosuppressed mice model. The results revealed that the Degree of Hydrolysis (DH) and percentage of soluble peptide content of the EPA were 22.38% and 77.92%, respectively. EPA was efficient in treating CTX-dropped mice's immunosuppression as well as intestinal inflammation. The peptide sequences of the EPA digestion products were identified, and ten top-scoring peptides were obtained using the PeptideRanker and HPepDOCK assessment, whose activity was validated in the macrophage model, with WNWLLPLMLG having the best immunomodulatory activity. Therefore, EPA may be a potential resource for immunomodulatory peptides, and autolysis can be a reliable technique for preparing peptides.

Effect of ethanol treatment on the structural, techno-functional, and antioxidant properties of edible insect protein obtained from Tenebrio molitor larvae

Edible insect-derived proteins have attracted considerable attention in the food industry owing to their excellent nutritional and bio-functional activities. Herein, ethanol (20, 40, 60, and 80 %)-treated Tenebrio molitor protein (ETMP) was prepared, and its structural, techno-functional, and antioxidant properties were assessed. As the ethanol concentration increased, the molecular weight of the ETMP decreased, and α-helix content decreased whereas that of β-sheet increased, affecting the secondary structure. Ethanol treatment also resulted in changes in the techno-functional properties of edible insect proteins. ETMP showed significant 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical-scavenging activity (p < 0.05), and its antioxidant activity effectively increased the viability of Vero cells damaged by oxidative stress; 20 % ethanol treatment induced the highest antioxidant activity. In conclusion, our results suggest that appropriate ethanol treatment (20 %) increases the antioxidant activity of edible insect proteins, suggesting their potential in food as an alternative protein resource and functional food with excellent antioxidant activity.

Effect of gamma-ray and electron-beam irradiation on the structural changes and functional properties of edible insect proteins from Protaetia brevitarsis larvae

Edible insects are regarded as future food sources to replace traditional livestock proteins. However, insect proteins have poor processing properties owing to various structural limitations. We investigated the structure of Protaetia brevitarsis larvae proteins modified by irradiation, and analyzed their resulting processing abilities. Following irradiation with gamma rays and electron beams, the ratio of parallel β-sheets to β-turns changed significantly, and changes in the tertiary protein structures were also confirmed. The polydispersity indices of the proteins remained relatively constant following irradiation, although the zeta potential and mean diameter changed. Furthermore, the pH, protein solubility, surface hydrophobicity, foaming capacity, and emulsion stability were higher than those of the control, whereas the viscosity and foaming stability were lower. Thus, gamma ray and electron beam irradiation clearly lead to changes in the structures of edible insect proteins and improves their processing properties, promoting the industrial utilization of such proteins in the food industry.

Structural, physicochemical, and immune-enhancing properties of edible insect protein isolates from Protaetia brevitarsis larvae.

Edible insects are promising future food resources globally. Herein, the structural, physicochemical, and bio-functional properties of edible insect protein isolates (EPIs) extracted from Protaetia brevitarsis larvae were investigated. The results showed that EPIs have a high total essential amino acid content; moreover, β-sheet is the major secondary protein structure. The EPI protein solution was highly soluble and electrically stable and did not aggregate easily. In addition, EPIs exhibited immune-enhancing properties; EPI treatment of macrophages induced the activation of macrophages and consequently promoted the production of pro-inflammatory mediators (NO, TNF-α, and IL-1β). Moreover, macrophage activation of EPIs was confirmed to occur through the MAPK and NF-κB pathways. In conclusion, our results suggest that the isolated P. brevitarsis protein can be fully utilized as a functional food material and alternative protein source in the future food industry.

Digestibility and quality of edible insect proteins: a systematic review ofin vivo studies

Edible insects are being considered as a potential sustainable food source to address global food security concerns and feed the world’s growing population. However, the quality and biological protein value of insects remain uncertain. This review analysedin vivo studies that evaluated edible insect consumption to clarify their potential as a high-quality protein source. Ten studies that evaluated protein quality in rats or mice were included, and most studies found weight gain similar to or greater than the control group. Food intake, net protein ratio, and biological value were also similar to those in the control group. However, true digestibility and protein efficiency ratio were generally lower in animals fed with insects, and higher nitrogen excretion was observed. The results of this systematic review were highly heterogeneous, and it cannot be concluded that insects have high protein quality, although positive results regarding weight gain and biological value were found.

One positive part of copper ion on biocatalysts: CNPs@Cu-Trypsin-assisted aqueous extraction technique for simultaneous exploration of edible insect oil and protein

Edible insects are rich in oil and protein resources, and has it become increasingly important to develop them reasonably when resources are increasingly scarce. However, due to the challenges in recovering enzymes in the enzyme-assisted aqueous extraction technique and the remaining organic solvents in the extraction process based on organic solvent, it is difficult to achieve simultaneous utilization of oil and protein from edible insects. Hence, an improved enzyme-assisted aqueous extraction process based on the assistance of chitosan cross-linked Fe3O4 nanoparticles (CNPs) immobilized enzymes and divalent copper ions chelation, CNPs@Cu-Trypsin-assisted aqueous extraction technique, was first proposed. The hydrolysis activity was significantly increased by 6.16 times, and the value of Kcat/Km was increased by 1.7 times for the results of the formation of the hydrophilic microenvironment. Furthermore, the extraction ratio of silkworm pupae oil extracted via CNPs@Cu-Trypsin-assisted aqueous extraction technique could be increased to 93%, and the oil extraction rate could be maintained above 65% after 10 times reused. Improvement of enzyme-assisted aqueous extraction under the addition of divalent copper ions could achieve an ideal effect of oil extracted from silkworm pupae with less time. Additionally, the quality of silkworm pupae oil was significantly improved and the silkworm pupae protein hydrolysate also showed improved biological activity. Therefore, the enzyme-assisted aqueous extraction enhancement by trypsin immobilization and divalent copper ion chelation is a promising strategy for the green and sustainable exploration of edible insect oil and protein without the use of organic solvents.

Alternative Protein Sources and Novel Foods: Benefits, Food Applications and Safety Issues.

The increasing size of the human population and the shortage of highly valuable proteinaceous ingredients has prompted the international community to scout for new, sustainable, and natural protein resources from invertebrates (e.g., insects) and underutilized legume crops, unexploited terrestrial and aquatic weeds, and fungi. Insect proteins are known for their nutritional value, being rich in proteins with a good balance of essential amino acids and being a valuable source of essential fatty acids and trace elements. Unconventional legume crops were found rich in nutritional, phytochemical, and therapeutic properties, showing excellent abilities to survive extreme environmental conditions. This review evaluates the recent state of underutilized legume crops, aquatic weeds, fungi, and insects intended as alternative protein sources, from ingredient production to their incorporation in food products, including their food formulations and the functional characteristics of alternative plant-based proteins and edible insect proteins as novel foods. Emphasis is also placed on safety issues due to the presence of anti-nutritional factors and allergenic proteins in insects and/or underutilized legumes. The functional and biological activities of protein hydrolysates from different protein sources are reviewed, along with bioactive peptides displaying antihypertensive, antioxidant, antidiabetic, and/or antimicrobial activity. Due to the healthy properties of these foods for the high abundance of bioactive peptides and phytochemicals, more consumers are expected to turn to vegetarianism or veganism in the future, and the increasing demand for such products will be a challenge for the future.

Ultrasound-assisted alkaline extraction of protein from Tenebrio molitor larvae: Extraction kinetics, physiochemical, and functional traits

Currently, as a promising alternative protein source, the interest of edible insect protein has been continuously increased. However, the extraction processing had distinct effects on the physicochemical properties and functionalities of this novel and sustainable protein. In this study, Tenebrio molitor larvae protein (TMLP) was extracted via ultrasound (US)-assisted alkaline extraction. The changes of extraction kinetics, physicochemical characteristics, and functional properties of TMLP as a function of US time (10, 20, 30, 40, 50 min) were investigated. The results showed that 30 min US treatment rendered the maximum protein yield (60.04 %) (P < 0.05). Meanwhile, Peleg's model was considered a suitable model to represent the extraction kinetics of TMLP, with a correlation coefficient of 0.9942. Moreover, the protein secondary structure, particle size, and amino acid profiles of TMLP were changed under the US-assisted alkaline extraction process. Additionally, a significant improvement of the functional properties of TMLP extracted with this method was observed compared to traditional alkaline extraction. In conclusion, the present work suggests that US-assisted alkaline extraction could be considered as a potential method to improve the protein yield, quality profiles, and functional properties of TMLP.

Protective effects of edible insect protein extracts from Protaetia brevitarsis against H2O2-induced oxidative stress in mouse C2C12 myoblast cells

Edible insects have been attracting significant attention not only for their nutritional superiority but also for their excellent bio-functional activity. In this study, the apoptotic protective effects of water-soluble protein (WPB) and salt-soluble protein (SPB) extracts derived from Protaetia brevitarsis against hydrogen peroxide (H2O2)-induced oxidative stress in C2C12 cells were investigated. WPB and SPB were confirmed to exhibit radical scavenging and ferric-ion reducing activities. In addition, WPB and SPB treatments effectively restored the decreased cell viability of the C2C12 cells induced by H2O2 treatment, and showcased protective effects against apoptosis and mitochondrial dysfunction by regulating the expressions of pro- and anti-apoptotic proteins. Therefore, our data indicated that protein extracts from Protaetia brevitarsis could be beneficial for the prevention and treatment of diseases associated with apoptosis induced by oxidative stress.

Current state of insect proteins: extraction technologies, bioactive peptides and allergenicity of edible insect proteins.

This review aims to provide an updated overview of edible insect proteins and the bioactivity of insect-derived peptides. The essential amino acid content of edible insects is compared with well-known protein sources to demonstrate that edible insects have the potential to cover the protein quality requirements for different groups of the population. Then the current methodologies for insect protein extraction are summarized including a comparison of the protein extraction yield and the final protein content of the resulting products for each method. Furthermore, in order to improve our understanding of insect proteins, their functional properties (such as solubility, foaming capacity, emulsifying, gelation, water holding capacity and oil holding capacity) are discussed. Bioactive peptides can be released according to various enzymatic hydrolysis protocols. In this context, the bioactive properties of insect peptides (antihypertensive, antidiabetic, antioxidant and anti-inflammatory properties) have been discussed. However, the allergens present in insect proteins are still a major concern and an unsolved issue for insect-based product consumption; thus, an analysis of cross reactivity and the different methods available to reduce allergenicity are proposed. Diverse studies of insect protein hydrolysates/peptides have been ultimately promoting the utilization of insect proteins for future perspectives and the emerging processing technologies to enhance the wider utilization of insect proteins for different purposes.

대체 단백질에 대한 소비자 인식 조사 분석: 배양육 및 식용곤충

The issue of protein alternatives to traditional livestock proteins is a policy that is in the spotlight as part of food security and carbon neutrality policies. This study was conducted to identify the degree of consumer awareness of alternative proteins through a consumer perception survey to support scientific progress of alternative proteins, and to suggest policies and technical directions accordingly. The survey was conducted on 500 adults over the age of 18 nationwide, centering on an online panel survey using a Structured Questionnaire. In this study, cultured meat and edible insect proteins were investigated among alternative proteins (cultured meat, edible insects, and plant-based proteins). Cultured meat was recognized by 64.6% and edible insects by 90.2%. Cultured meat showed higher interest than edible insects, and cultured meat showed high selection in terms of resource saving and environment protection. Edible insects received many choices in terms of economical value to secure the food. The opinion that it is necessary to develop cultured meat rather than edible insect protein was high in Korea. As a result of this study, it was confirmed that the recognition of the alternative protein was high, and in particular, it was confirmed that the need for cultured meat technology was quite high.

Recent Insight on Edible Insect Protein: Extraction, Functional Properties, Allergenicity, Bioactivity, and Applications.

Due to the recent increase in the human population and the associated shortage of protein resources, it is necessary to find new, sustainable, and natural protein resources from invertebrates (such as insects) and underutilized plants. In most cases, compared to plants (e.g., grains and legumes) and animals (e.g., fish, beef, chicken, lamb, and pork), insect proteins are high in quality in terms of their nutritional value, total protein content, and essential amino acid composition. This review evaluates the recent state of insects as an alternative protein source from production to application; more specifically, it introduces in detail the latest advances in the protein extraction process. As an alternative source of protein in food formulations, the functional characteristics of edible insect protein are comprehensively presented, and the risk of allergy associated with insect protein is also discussed. The biological activity of protein hydrolyzates from different species of insects (Bombyx mori, Hermetia illucens, Acheta domesticus, Tenebrio molitor) are also reviewed, and the hydrolysates (bioactive peptides) are found to have either antihypertensive, antioxidant, antidiabetic, and antimicrobial activity. Finally, the use of edible insect protein in various food applications is presented.