Single Cell Protein Production Research Articles

Towards a Biorefinery Processing Waste from Plantain Agro-Industry: Process Design and Techno-Economic Assessment of Single-Cell Protein, Natural Fibers, and Biomethane Production through Process Simulation

The plantain agro-industry generates different residues in the harvest and post-harvest stages. Therefore, the design of processes for valorization is required. The aim of this work was to design and techno-economically evaluate the processes for the production of single-cell protein, natural fibers, and biomethane from plantain residues by process simulation in the framework of the design of a future biorefinery for valorization of these residues. The processes were simulated using SuperPro Designer. The scale size was calculated at 1,267,071 tons for the processing of plantain lignocellulosic waste (pseudostems) and 3179 tons for the processing of starchy waste (rejected unripe plantain fruits). The results obtained suggest that the best alternative for the valorization of plantain residues corresponded to the production of natural fibers, with a net present value of $29,299,000. This work shows that waste from the plantain agro-industry exhibits high potential as a feedstock for the production of value-added products. In addition, the process flowsheets simulated in this work can be integrated into the basic design of a biorefinery processing plantain waste.

Recycling of acetate and ammonium from digestate for single cell protein production by a hybrid electrochemical-membrane fermentation process

Digestate is a potential substrate for yeast to produce single cell protein (SCP), but its complex matrix constrains applications. This study developed a hybrid electrochemical-membrane fermentation process to recover nutrients from synthetic digestate and produce yeast SCP. Electrodialysis (ED) was first employed to recover acetate and ammonium. More nutrients were recovered at a higher voltage but with lower current efficiency. At 2 V, 42.2% and 60.1% of acetate and ammonium were reclaimed, and further concentrated by 14- and 10-fold via a forward osmosis (FO) process. Due to the complexity of real digestate, the recovery efficiency of ED in practical applications should be further investigated. Saccharomyces cerevisiae fed on raw and concentrated recovery achieved similar SCP productions of 0.76–0.86 g/L, with amino acid concentrations above the FAO recommendation by 1.5–4.2-fold. The total production cost was estimated to be €20.4/kg-SCP by single ED, while the hybrid ED-FO process cost 40 times more.

Production of Single-Cell Protein from Fruit and Vegetable Waste Using an Isolated Strain of Lactobacillus sp.

Production of Single-Cell Protein from Fruit and Vegetable Waste Using an Isolated Strain of <i>Lactobacillus sp.</i>

Yeast community composition impacts on tequila industry waste treatment for pollution control and waste-to-product synthesis

The Tequila industry is a major producer of wastewater in the state of Jalisco, Mexico. Tequila vinasses (TV) are a residue from the distillation of fermented agave wort during tequila production. TV are difficult to treat due their high organic content, high nutrient loads, acidic pH and high discharge temperature. TV are frequently released into waterbodies or soil without any treatment, leading to environmental degradation of soil and water sources. To reduce the environmental impact of the tequila industry, cost-effective TV revalorization approaches must be developed. The goals of the present study were to assess the treatment and revalorization potential of TV using mono and mixed yeast cultures to produce single-cell protein (SCP) and to analyze yeast community composition using high-throughput sequencing during the mixed-culture fermentation of TV. The fermentation process was performed using a mixed culture of three fodder yeast species (Candida utilis, Rhodotorula mucilaginosa and Kluyveromyces marxianus) during 48 h at benchtop-scale. High-throughput sequencing was performed to assess the relative abundance of the yeast communities. Additionally, a redundancy analysis was performed to analyze the bidirectional influence between yeast communities and pollutant removal (COD, nitrogen, phosphorus, proteins, and sugars). Mixed yeast cultures displayed overall higher pollutant removal rates than monocultures, where C. utilis and K. marxianus contributed the most to pollutant removal and protein accumulation. The R. mucilaginosa population declined rapidly in mixed culture, presumably due to TV acidity and phenolic composition. However, the presence of The R. mucilaginosa in the mixed culture enhanced pollutant removal and amino acid contents. Accordingly, the protein and amino acid content within mixed cultures were significantly higher than those of monocultures, indicating that mixed cultures have a strong potential to produce protein rich biomass from TV, aiding in the transition of both the tequila and the livestock industries to a sustainable circular bioeconomy model by the reintegration of organic material flows into productive processes, reducing raw resource intake and waste generation. The present circular bioeconomy approach could represent a potential to produce 45,664 tons of protein feed yearly, based on the current tequila vinasses generated in the state of Jalisco.

Isolation, Molecular Identification and Amino Acid Profiling of Single-Cell-Protein-Producing Phototrophic Bacteria Isolated from Oil-Contaminated Soil Samples.

In the current study, soil samples were gathered from different places where petrol and diesel filling stations were located for isolation of photosynthetic bacteria under anaerobic conditions using the paraffin wax-overlay pour plate method with Biebl and Pfennig’s medium. The three isolated strains were named Rhodopseudomonas palustris SMR 001 (Mallapur), Rhodopseudomonas palustris NR MPPR (Nacahram) and Rhodopseudomonas faecalis N Raju MPPR (Karolbagh). The morphologies of the bacteria were examined with a scanning electron microscope (SEM). The phylogenetic relationship between R. palustris strains was examined by means of 16S rRNA gene sequence analysis using NCBI-BLAST search and a phylogenetic tree. The sequenced data for R. palustris were deposited with the National Centre for Biotechnology Research (NCBI). The total amino acids produced by the isolated bacteria were determined by HPLC. A total of 14 amino acids and their derivatives were produced by the R. palustris SMR 001 strain. Among these, carnosine was found in the highest concentration (8553.2 ng/mL), followed by isoleucine (1818.044 ng/mL) and anserine (109.5 ng/mL), while R. palustris NR MPPR was found to produce 12 amino acids. Thirteen amino acids and their derivatives were found to be produced from R. faecalis N Raju MPPR, for which the concentration of carnosine (21601.056 ng/mL) was found to be the highest, followed by isoleucine (2032.6 ng/mL) and anserine (227.4 ng/mL). These microbes can be explored for the scaling up of the process, along with biohydrogen and single cell protein production.

Optimizing medium composition with wastewater from Coffea arabica processing to produce single‐cell protein using Candida sorboxylosa

AbstractBackgroundCoffee production is an important agro‐industrial activity in several countries worldwide. However, during wet processing of coffee beans, substantial quantities of wastewater effluents are generated and discharged into nearby water systems, negatively impacting ecosystems. A compelling biotechnological alternative involves the use of nutrients contained in wastewater (e.g., sugars, proteins, and salts) to produce value‐added molecules–such as single‐cell protein (SCP) using the yeast Candida sorboxylosa–with high potential for use as animal feed.ResultsYeast isolated from the feces of ring‐tailed coatis showed a positive assimilation of reducing sugars (glucose, mannose, and fructose). Efficient reuse of coffee wastewater (CWW) for SCP production was achieved via the statistical design of experiments (DoE) technique. The bioprocess was standardized in a medium culture composed of 87.5% (v/v) CWW, and of 1.38 and 7.24 g/L yeast extract (YE) and ammonium sulfate [(NH4)2SO4], respectively, resulting in a high SCP yield of approximately 38%.ConclusionThe reuse of CWW shows potential as a low‐cost component for SCP production that can ultimately reduce the environmental impact of wastewater effluent by circumventing its release into river systems. The bioprocess developed in this study presented certain advantages, including high volumes of CWW with only YE and (NH4)2SO4 supplementation, and without requiring additional components to correct the pH of the medium. Therefore, it is highly feasible to apply this bioprocess in close proximity to coffee‐processing agrobusinesses, thus generating an alternative source of income for Peruvian coffee‐producing communities. © 2022 Society of Chemical Industry (SCI).

Single-Cell Proteins Obtained by Circular Economy Intended as a Feed Ingredient in Aquaculture.

The constant increment in the world’s population leads to a parallel increase in the demand for food. This situation gives place the need for urgent development of alternative and sustainable resources to satisfy this nutritional requirement. Human nutrition is currently based on fisheries, which accounts for 50% of the fish production for human consumption, but also on agriculture, livestock, and aquaculture. Among them, aquaculture has been pointed out as a promising source of animal protein that can provide the population with high-quality protein food. This productive model has also gained attention due to its fast development. However, several aquaculture species require considerable amounts of fish protein to reach optimal growth rates, which represents its main drawback. Aquaculture needs to become sustainable using renewable source of nutrients with high contents of proteins to ensure properly fed animals. To achieve this goal, different approaches have been considered. In this sense, single-cell protein (SCP) products are a promising solution to replace fish protein from fishmeal. SCP flours based on microbes or algae biomass can be sustainably obtained. These microorganisms can be cultured by using residues supplied by other industries such as agriculture, food, or urban areas. Hence, the application of SCP for developing innovative fish meal offers a double solution by reducing the management of residues and by providing a sustainable source of proteins to aquaculture. However, the use of SCP as aquaculture feed also has some limitations, such as problems of digestibility, presence of toxins, or difficulty to scale-up the production process. In this work, we review the potential sources of SCP, their respective production processes, and their implementation in circular economy strategies, through the revalorization and exploitation of different residues for aquaculture feeding purposes. The data analyzed show the positive effects of SCP inclusion in diets and point to SCP meals as a sustainable feed system. However, new processes need to be exploited to improve yield. In that direction, the circular economy is a potential alternative to produce SCP at any time of the year and from various cost-free substrates, almost without a negative impact.

Production of Single Cell Protein from Fruit Waste

Abstract: The fast growing population of the world has led to an increased demand for protein rich foods. The conversion of fruit waste into single cell protein is an alternative solution to the worldwide protein deficiency and environmental pollution concerns. Dried cells of bacteria, algae, yeast and fungi which are rich in protein and could be used as dietary supplements are called single cell protein. In the present study single cell protein represent microbial cells grown in mass culture and harvested for use as protein source in foods and animal feed. Fruit waste was used as sole carbon source for preparation of fermentation on used for the growth of bacteria, fungi, algae and yeast. The increased incubation time of different fruits hydrolaysate enhanced the biomass yield and the protein formation within the cell. The bioconversion of fruit waste by microbial fermentation into single cell protein has potential use for solving the worldwide protein deficiency in food. The finding in this study indicated that fruit waste could serve as a good substrate for single cell protein production.

Distributive and collaborative push‐and‐pull in an artificial microbial consortium for improved consolidated bioprocessing

AbstractThe consolidated bioprocessing (CBP) of lignocellulose by the synthetic microbial consortium of Trichoderma reesei and Saccharomyces cerevisiae is a promising way of biomanufacturing d‐glucaric acid. However, the hindrance factor to its industrial application is the low efficiency. Therefore, we engineered T. reesei and S. cerevisiae to improve the CBP for d‐glucaric acid production. T. reesei was engineered to produce more cellulase and release more fermentable sugars from lignocellulose, that is, pushing more sugars to S. cerevisiae. S. cerevisiae was engineered to metabolize cellobiose and siphon more sugars into d‐glucaric acid biosynthetic pathway, that is, pulling more sugars to S. cerevisiae. This is the strategy of distributive and collaborative push‐and‐pull we developed and proposed in this work, which was proven successful in improving efficiencies of the CBPs of steam‐exploded corn stover (SECS) for d‐glucaric acid production and distiller's grains for single cell protein (SCP) production. The titer, yield and productivity of d‐glucaric acid produced from 50 g/L SECS by the microbial consortium of T. reesei C10 and S. cerevisiae LGA‐1C3S2 were 6.42 g/L, 0.128 g/g SECS, and 0.917 g/L/d, respectively. The titer, yield, and productivity of SCP produced from 80 g/L distiller's grains were 50.5 g/L, 0.631 g/g distiller's grains, and 8.417 g/L/d, respectively. These were much higher than the initial microbial consortia of T. reesei Rut‐C30 and S. cerevisiae LGA‐1 or INVSc1. The results confirmed the applicability and generalizability of distributive and collaborative push‐and‐pull, which has profound meaning for science and engineering.

Single Cell Protein for Foods and Feeds: A Review of Trends

Introduction:Predictions on the world’s population in the next few decades suggest that the global demand for animal-derived proteins may not be met if current conventional agriculture approaches are used. One promising solution to this complex crisis lies in the use of single-cell proteins (SCP). SCP refers to the edible biomass of unicellular microorganisms and can be developed as animal feeds or human foods. This paper provides a detailed overview on research towards the production and utilisation of SCPs and trends within the field.Study Design:A bibliometric based study was conducted on 425 SCP research articles collected from the Web of Science database, analysing the most cited papers using VOSviewer software, and contributing authors, affiliations and country of origin. Research publications on SCP started in 1961 and has grown steadily over the years.Discussion:Emerging research topics within SCP production focused on the use of improved fungal strains, the composition and characteristics of SCPs based on the type of substrates used, industrial production processes and the use of waste for SCP production, which serves the dual purpose of mitigating the cost associated with waste disposal and production of a valuable product.

Single cell protein production by Candida robusta isolated from sugar cane (Saccharum sp.) for animal feed

The protein obtained from the microorganisms is not only cheap but can be used as additive to provide a balanced nutrition for many animals feeding. The aim of this work was the single cell protein (SCP) production by Candida robusta URM5293 using sugarcane bagasse as substrate. The yeast C. robusta URM5293 was isolated from root sugarcane and was identified based on morphological and biochemical characteristics. Biomass production was done into Erlenmayers flasks (250 mL) containing culture medium supplemented of sugarcane bagasse hydrolyzate. Fermentations were carried varying culture conditions through the study of four different variables: temperature (25, 30, and 35°C), agitation intensity (110, 140 or 170 rpm), pH (6.0, 7.0, and 8.0) and production time (72, 96, and 120h), according to a 24-1 fractional factorial design. Results demonstrated that this yeast was able to ensure the highest level of biomass (141 g/L) when cultivated at 25°C, pH 6.0, 170 rpm of agitation intensity after 72h of cultivation using sugarcane bagasse. The present results demonstrate the potential of sugarcane bagasse hemicellulosic hydrolyzate as a substrate for the production of microbial protein by C. robusta URM5293.

Upcycling waste organic acids and nitrogen into single cell protein via brewer's yeast

The food crisis due to the global population boom requires agriculture revolutions and a transition to sustainable, affordable sources of food proteins. Single cell protein (SCP) derived from microorganisms such as brewer's yeast is one of the best possible options to meet the protein challenges. Utilization of carbon and nitrogen recovered from waste streams, instead of sugar-rich substrate, for yeast-driving SCP production could improve the overall sustainability from both economic and environmental points of view. The lack of a comprehensive understanding of the process such as the interactions between different organic acids/ions and SCP production by yeast (e.g., Saccharomyces cerevisiae) has placed a major impediment to the valorization effort. This study tackles this challenge by exploring the impact of various organic acids, nitrogen and ionic species on SCP production by S. cerevisiae. It was found that the SCP production increased with acetate concentrations (2.6–20 g-COD/L). A maximal protein concentration of 0.75 g/L was harvested using 20 g-COD/L acetate. Intermittent acid addition for pH neutralization further enhanced SCP production by 67% at 0.94 g/L. Ammonia was the best inorganic nitrogen source for S. cerevisiae among nitrate and nitrite. S. cerevisiae was not able to utilize lactate, propionate and butyrate alone for growth, but it can catabolize lactate along with acetate. The addition of oleate and linoleate resulted in a significant increase in the protein content of biomass by 1.8- and 1.6-fold, respectively. S. cerevisiae using acetate required eight times fewer K+ than Na+. Typical anions including Cl−, NO3− and SO42− (20–500 mM) were not found harmful for SCP synthesis. The results help to fill the knowledge gap in yeast-driving waste-to-protein conversion process, bring high impacts and positive implications for waste valorization and future feed and food supply.

The Use of “Organic Protein” in Feed Formulation on Intestinal and Ammonia Levels of Broiler

The aim of this study is to determine the effect of Organic Protein as a feed ingredient on pH, viscosity and enzyme activity in the small intestine and levels of ammonia in broiler excreta. Organic Protein is a Single Cell Protein (SCP) product name from PT. Miwon Indonesia that has a protein content up to 40%. We conducted a field experiment and used DOC broilers and Organic Protein as treatment feed in a Completely Randomized Design (CRD) comprising of 5 treatments and 6 replicates, accounting for 30 experimental units. The feed treatments were basal feed T0, T1, T2, T3 and T4 added with Organic Protein 0%, 1.5%, 3%, 4.5%, and 6%, respectively. The observed variables were pH, viscosity, enzyme activity (amylase and protease) of small intestine and excreta ammonia level of broiler. The data were analyzed with ANOVA followed by Duncan’s Multiple Range Test (DMRT). The results showed the treatments have very significantly different effects (P&lt;0.01) on the amylase and protease activities, but significantly different effects (P&gt;0.05) on pH, viscosity of small intestine, and ammonia levels of excreta. The use of Organic Protein at a level of 4.5% gave the best results to increase enzyme activity of amylase and protease.

Valorization of food waste fermentation liquid into single cell protein by photosynthetic bacteria via stimulating carbon metabolic pathway and environmental behaviour

Single cell protein (SCP) production by photosynthetic bacteria (PSB) is dependent on the bioavailability of carbon source, while sufficient volatile fatty acids (VFAs) in food waste fermentation liquid might be a potential alternative. It is unclear how the fermentation liquid affects the SCP biosynthesis and the related metabolic mechanism. This work demonstrated that the SCP production could be improved effectively (2088.4 mg/L) with high conversion capacity of carbon source (0.99 mg-biomass/mg-COD) by regulating carbon source level. PSB preferred to utilize the VFAs in food waste fermentation liquid. The carbon metabolic pathways (e.g., the transformation of VFAs to acetyl-CoA, and tricarboxylic acid cycle) involved in the SCP production were enhanced under optimal condition. Moreover, optimal carbon source regulation could significantly stimulate the environmental behaviour of PSB (e.g., two-component system, quorum sensing, and ATP-binding cassette transporter) involved in adaptation to external stimulus and maintaining high bacterial activity, resulting in SCP yield promotion.

Production of Single-Cell Protein from Fruit Peel Wastes Using Palmyrah Toddy Yeast

Single-cell protein (SCP) from agro-waste material has gained increased attention in the recent past as a relatively cheap and alternative protein source to meet the nutritional demand generated by the fast-growing population. Furthermore, bioconversion of these wastes into SCP such as value-added products reduce the environmental-related issues. In this study, locally available pineapple (Ananas comosus), watermelon (Citrullus lanatus), papaya (Carica papaya), sour orange (Citrus medica), banana (Musa acuminata) and mango (Mangifera indica) peel wastes were investigated for their suitability to produce SCP using palmyrah (Borassus flabellifer) toddy carrying natural mixed yeast and bacteria culture under liquid state fermentation system. Moreover, this study attempted to select the best substrate and the optimized process condition for SCP production to increase the protein yield. The physicochemical properties of selected fruit peels were analyzed. The sterilized peel extracts (10%, v/v) were inoculated with 5 mL of palmyrah toddy and allowed to ferment in a shaking incubator at 100 rpm for 48 h in triplicate At the end of fermentation, the sediments were collected by centrifugation at 1252× g, oven-dried, and the dry weight was taken to determine the protein content. The biomass yield ranged from 5.3 ± 0.6 to 11.7 ± 0.8 g/L, with the least biomass yield being observed with watermelon peels while the maximum yield was observed with papaya peels. Papaya peel generated a significantly higher (p &lt; 0.05) amount of protein (52.4 ± 0.4%) followed by pineapple (49.7 ± 1.3%), watermelon (45.2 ± 0.7%), banana (30.4 ± 0.6%), sour orange (29.5 ± 1.2%) and mango (24.6 ± 0.2%) peels. The optimum condition for the fermentation of papaya waste was pH 5.0, 25 °C, and 24 h. Nucleic acid reduction treatment significantly reduces dry weight and protein content of biomass. It can be concluded that papaya peel waste is a suitable substrate for protein-rich cell biomass production using the natural toddy mixed culture of palmyrah.

Two-stage fermentation enhanced single-cell protein production by Yarrowia lipolytica from food waste

The resource utilization of food waste is crucial, and single-cell protein (SCP) is attracting much attention due to its high value. This study aimed to convert food waste to SCP by Yarrowia lipolytica. It was found the chemical oxygen demand (COD) removal rate 77 ± 1.70% was achieved at 30 g COD/L with the protein content of biomass only 24.1 ± 0.4% w/w biomass dry weight (BDW) in one-stage fermentation system. However, the protein content was significantly increased to 38.8 ± 0.2% w/w BDW with the COD removal rate 85.5 ± 0.7% by a two-stage fermentation process, where the food waste was firstly anaerobically fermented to volatile fatty acids and then converted to SCP with Yarrowia lipolytica. Transcriptomic analysis showed that the expression of SCP-producing genes including ATP citrate (pro-S)-lyase and fumarate hydratase class II were up-regulated in the two-stage transformation, resulting in more organic degradation for SCP synthesis.

Single cell protein production by Candida robusta isolated from sugar cane (Saccharum sp.) for animal feed / Produção de Biomassa por Candida robusta isolada da Cana-de-açúcar (Saccharum sp.) para alimentação animal

The protein obtained from the microorganisms is not only cheap but can be used as additive to provide a balanced nutrition for many animals feeding. The aim of this work was the single cell protein (SCP) production by Candida robusta URM5293 using sugarcane bagasse as substrate. The yeast C. robusta URM5293 was isolated from root sugarcane and was identified based on morphological and biochemical characteristics. Biomass production was done into Erlenmayers flasks (250 mL) containing culture medium supplemented of sugarcane bagasse hydrolyzate. Fermentations were carried varying culture conditions through the study of four different variables: temperature (25, 30, and 35°C), agitation intensity (110, 140 or 170 rpm), pH (6.0, 7.0, and 8.0) and production time (72, 96, and 120h), according to a 24-1 fractional factorial design. Results demonstrated that this yeast was able to ensure the highest level of biomass (141 g/L) when cultivated at 25°C, pH 6.0, 170 rpm of agitation intensity after 72h of cultivation using sugarcane bagasse. The present results demonstrate the potential of sugarcane bagasse hemicellulosic hydrolyzate as a substrate for the production of microbial protein by C. robusta URM5293.

Single Cell Protein Production Using Different Fruit Waste: A Review

The single cell protein (SCP) technique has become a popular technology in recent days, which addresses two major issues: increasing world protein deficiency with increasing world population and the generation of substantial industrial wastes with an increased production rate. Global fruit production has increased over the decades. The non-edible parts of fruits are discarded as wastes into the environment, which may result in severe environmental issues. These fruit wastes are rich in fermentable sugars and other essential nutrients, which can be effectively utilized by microorganisms as an energy source to produce microbial protein. Taking this into consideration, this review explores the use of fruit wastes as a substrate for SCP production. Many studies reported that the wastes from various fruits such as orange, sweet orange, mango, banana, pomegranate, pineapple, grapes, watermelon, papaya, and many others are potential substrates for SCP production. These SCPs can be used as a protein supplement in human foods or animal feeds. This paper discusses various aspects in regard to the potential of fruit wastes as a substrate for SCP production.

Microbial conversion of syngas to single cell protein: The role of carbon monoxide

Syngas has been widely utilized as substrate in microbial processes to produce various high-value products. Nevertheless, its applicability for single cell protein (SCP) production by hydrogen-oxidizing bacteria (HOB) has never been explored. Especially, the role of carbon monoxide in SCP synthesis is unknown. Thus, this study first investigated the effect of different CO and H2 ratios in syngas on Cupriavidus necator H16 growth and SCP production. The growth of C. necator H16 was significantly restrained with the increase of CO content, and it almost ceased once the CO:H2 ratios were above 1. In addition, the growth rate of C. necator H16 increased with the increase of shaking frequency and inoculum volume. Furthermore, larger gas–liquid interfacial areas and gas input amount resulted in a higher growth rate and OD value. The raw protein contents were around 50–60% regardless of CO concentration, and the amino acid profiles showed no apparent differences. The results showed that although the presence of CO led to the reduction of the growth rate of C. necator H16, it had a limited effect on the SCP quality. H2 and CO2 in syngas were the primary substrates for SCP production, while CO served more as a toxic inhibitor. The successful demonstration of SCP synthesis from syngas may provide a superior route for the valorization of waste biomass by turning it into food or feed.

Protein Sources Alternative to Meat: State of the Art and Involvement of Fermentation.

Meat represents an important protein source, even in developing countries, but its production is scarcely sustainable, and its excessive consumption poses health issues. An increasing number of Western consumers would replace, at least partially, meat with alternative protein sources. This review aims at: (i) depicting nutritional, functional, sensory traits, and critical issues of single-cell proteins (SCP), filamentous fungi, microalgae, vegetables (alone or mixed with milk), and insects and (ii) displaying how fermentation could improve their quality, to facilitate their use as food items/ingredients/supplements. Production of SCP (yeasts, filamentous fungi, microalgae) does not need arable land and potable water and can run continuously, also using wastes and byproducts. Some filamentous fungi are also consumed as edible mushrooms, and others are involved in the fermentation of traditional vegetable-based foods. Cereals, pseudocereals, and legumes may be combined to offer an almost complete amino acid profile. Fermentation of such vegetables, even in combination with milk-based products (e.g., tarhana), could increase nutrient concentrations, including essential amino acids, and improve sensory traits. Different insects could be used, as such or, to increase their acceptability, as ingredient of foods (e.g., pasta). However, insects as a protein source face with safety concerns, cultural constraints, and a lack of international regulatory framework.