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Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications

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A special issue of Foods (ISSN 2304-8158). This special issue belongs to the section "Food Security and Sustainability".

Deadline for manuscript submissions: closed (15 December 2023) | Viewed by 54916

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Dr. Epameinondas Xanthakis

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Guest Editor

Department of Food Science and Technology, School of Food Sciences, University of West Attica, Egaleo, Greece
Interests: food production side-stream valorization; alternative proteins; food freezing; food processing; food quality; food chemistry; microwaves; enzymology; phytochemicals
Special Issues, Collections and Topics in MDPI journals

Dr. Charilaos Xiros

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RISE Processum AB, Box 70, SE-891 22 Örnsköldsvik, Sweden
Interests: industrial microbiology; biotechnology; alternative proteins; single cell proteins

Special Issue Information

Dear Colleagues,

Traditional animal-based sources offer food proteins with high nutritional value. Despite their high nutritional and functional properties, their production has been characterized as unsustainable due to their increased environmental impact. Global human population rapid growth over the last century and the predictions for the coming years raise the need for increased food protein production. The continuous population growth in combination with climate change and the environmental risks that our planet face gave a nudge to policymakers and society to explore novel, alternative, and sustainable food proteins which may efficiently substitute the traditional animal-based ones.

Academia and industry have been working actively on this scope over the last few years, seeking novel protein sources with high nutritional value, low environmental impact, high production yields, cost efficiency, and desired technofunctional and sensory properties to be used as ingredients in food products. Ocean-based proteins and microalgae, fungi, plant-based proteins, cultured meat, and insects are the most developed solutions to date. This Special Issue will gather basic and applied research from academia and industry focused on a wide range of novel terrestrial, marine, and biotechnological sources for food applications, selecting relevant original research and review articles which will offer insights and solutions to the current scientific and societal challenges.

Dr. Epameinondas Xanthakis
Dr. Charilaos Xiros
Guest Editors

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Keywords

alternative proteins
single cell protein
plant-based protein
insect
food processing
food structure
food quality
digestibility
allergenicity

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Published Papers (11 papers)

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12 pages, 1816 KiB

Open AccessArticle

Effect of Crude Polysaccharides from Ecklonia cava Hydrolysate on Cell Proliferation and Differentiation of Hanwoo Muscle Stem Cells for Cultured Meat Production

by Jae-Hoon Lee, Tae-Kyung Kim, Min-Cheol Kang, Min-Kyung Park, Sang-Hun Park, Jung-Seok Choi and Yun-Sang Choi

Foods 2024, 13(4), 563; https://doi.org/10.3390/foods13040563 - 13 Feb 2024

Viewed by 1535

Abstract

Ecklonia cava, a brown seaweed native to the East Asian coast, is known for its unique composition, including polysaccharides, polyphenols, and phlorotannins. Fucoidan is a sulfated polysaccharide widely used as a functional ingredient in foods. This study obtained crude polysaccharides (ECC_CPS) from E. cava celluclast enzymatic hydrolysate using ethanol precipitation. ECC_CPS increased cell viability during the proliferation of Hanwoo muscle satellite cells (HMSCs). The effect of ECC_CPS on the expression of proliferation-related markers was confirmed as MYF5 and MYOD expression significantly increased, whereas PAX7 expression was maintained. The evaluation of cell migration activity has a major impact on cell proliferation and differentiation, and the cell migration index significantly increased with ECC_CPS treatment (p < 0.01). This was related to the HGF/MET pathway and FAK pathway. Treatment with ECC_CPS promoted differentiation at the cell differentiation stage, thereby increasing the expression of differentiation markers, such as MYH2, MYH7, and MYOG (p < 0.001 or p < 0.01). Therefore, our findings imply that crude polysaccharide obtained from E. cava can be an additive ingredient that enhances the proliferation and differentiation of muscle satellite cells used in the manufacture of cultured meat products. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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18 pages, 5627 KiB

Open AccessArticle

Future Proteins: Sustainable Diets for Tenebrio molitor Rearing Composed of Food By-Products

by Andrea Lienhard, René Rehorska, Barbara Pöllinger-Zierler, Chiara Mayer, Monika Grasser and Simon Berner

Foods 2023, 12(22), 4092; https://doi.org/10.3390/foods12224092 - 11 Nov 2023

Cited by 9 | Viewed by 2871

Abstract

Since the human population is continuously growing, sufficient food with low environmental impact is required. Especially, the challenge of providing proteins will deepen and insects can contribute to a more sustainable and efficient source of protein for human consumption. Tenebrio molitor larvae are highly nutritious and rearing mealworms is more environmentally friendly compared to the production of traditional livestock meat. To use T. molitor as a more sustainable alternative to conventional proteins, it is essential to apply diets from a local and sustainable source. Therefore, the objective of this study was to find local by-products or leftovers which can be used in mass production of larvae as a main substrate. Feeding trials investigating twenty-nine different substrates were conducted to evaluate larval growth performance and adult reproduction by determining development times, survival rates, biomass, and fecundity. Several suitable by-products were identified that can be used in high quantities as single component diet for T. molitor rearing, revealing a high survival rate, short development time, high mean total biomass, and successful breeding. The most successful substrate—malt residual pellets—was found to be an alternative to the most used substrate, wheat bran. Furthermore, corn germ meal, sweet chestnuts, bread remains, soybeans, sweet potatoes, and wheat germs have been discovered to be suitable diets for T. molitor. Moreover, the findings of this study contribute towards using several substrates as supplements. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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19 pages, 4225 KiB

Open AccessArticle

Characterisation of the Enzymatically Extracted Oat Protein Concentrate after Defatting and Its Applicability for Wet Extrusion

by Darius Sargautis, Tatjana Kince and Ilze Gramatina

Foods 2023, 12(12), 2333; https://doi.org/10.3390/foods12122333 - 10 Jun 2023

Cited by 3 | Viewed by 1843

Abstract

An oat protein concentrate (OC1) was isolated from oat flour through starch enzymatic hydrolysis, by subsequent defatting by ethanol and supercritical fluid extraction (SFE) reaching protein concentrations of 78% and 77% by weight in dry matter, respectively. The protein characterisation and functional properties of the defatted oat protein concentrates were evaluated, compared and discussed. The solubility of defatted oat protein was minor in all ranges of measured pH (3–9), and foamability reached up to 27%. Further, an oat protein concentrate defatted by ethanol (ODE1) was extruded by a single screw extruder. The obtained extrudate was evaluated by scanning electron microscope (SEM), texture and colour analysers. The extrudate’s surface was well formed, smooth, and lacking a tendency to form a fibrillar structure. Textural analysis revealed a non-unform structure (fracturability 8.8–20.9 kg, hardness 26.3–44.1 kg) of the oat protein extrudate. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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18 pages, 2721 KiB

Open AccessArticle

Mycoprotein Production by Submerged Fermentation of the Edible Mushroom Pleurotus ostreatus in a Batch Stirred Tank Bioreactor Using Agro-Industrial Hydrolysate

by Georgios Bakratsas, Angeliki Polydera, Oskar Nilson, Alexandra V. Chatzikonstantinou, Charilaos Xiros, Petros Katapodis and Haralambos Stamatis

Foods 2023, 12(12), 2295; https://doi.org/10.3390/foods12122295 - 7 Jun 2023

Cited by 16 | Viewed by 5079

Abstract

The demand for cheap, healthy, and sustainable alternative protein sources has turned research interest into microbial proteins. Mycoproteins prevail due to their quite balanced amino acid profile, low carbon footprint and high sustainability potential. The goal of this research was to investigate the capability of Pleurotus ostreatus to metabolize the main sugars of agro-industrial side streams, such as aspen wood chips hydrolysate, to produce high-value protein with low cost. Our results indicate that P. ostreatus LGAM 1123 could be cultivated both in a C-6 (glucose)- and C-5(xylose)-sugar-containing medium for mycoprotein production. A mixture of glucose and xylose was found to be ideal for biomass production with high protein content and rich amino acid profile. P. ostreatus LGAM 1123 cultivation in a 4 L stirred-tank bioreactor using aspen hydrolysate was achieved with 25.0 ± 3.4 g L⁻¹ biomass production, 1.8 ± 0.4 d⁻¹ specific growth rate and a protein yield of 54.5 ± 0.5% (g/100 g sugars). PCA analysis of the amino acids revealed a strong correlation between the amino acid composition of the protein produced and the ratios of glucose and xylose in the culture medium. The production of high-nutrient mycoprotein by submerged fermentation of the edible fungus P. ostreatus using agro-industrial hydrolysates is a promising bioprocess in the food and feed industry. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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13 pages, 1698 KiB

Open AccessArticle

Growth Optimization and Rearing of Mealworm (Tenebrio molitor L.) as a Sustainable Food Source

by Kanwal Riaz, Toheed Iqbal, Sarzamin Khan, Amjad Usman, Mariam S. Al-Ghamdi, Ashwag Shami, Rania Ali El Hadi Mohamed, Abdulrahman A. Almadiy, Fahd Mohammed Abd Al Galil, Nawal Abdulaziz Alfuhaid, Nazeer Ahmed and Pravej Alam

Foods 2023, 12(9), 1891; https://doi.org/10.3390/foods12091891 - 4 May 2023

Cited by 11 | Viewed by 5524

Abstract

As a sustainable food source for humans, mealworms (Tenebrio molitor) have a great deal of potential, due to the fact that they have a very favorable nutritional profile and a low environmental impact. For meal production, feed formulation and optimization are important. The mealworm Tenebrio molitor (Coleoptera: Tenebrionidae) is the most consumed insect in the world. Mealworms were given a variety of diets, including wheat bran as constant diet supplemented with different levels of Ospor (Bacillus clausii) at 0.002 g, 0.004 g, 0.006 g, and 0.008 g; imutec (Lacticaseibacillus rhamnosus) at 0.2 g. 0.4 g, 0.6 g, and 0.8 g; fungi (Calocybe indica) at 250 g, 500 g, and 750 g; yeast (Saccharomyces cerevisiae) at 50 g, 100 g, and 150 g; and wheat bran (standard diet) were examined in complete randomized design (CRD). Different parameters, i.e., the larval, pupal, and adult weight, size, life span, and nutritional profile of mealworm were studied. When compared with other insect growth promoters, only wheat bran was discovered to be the most efficient. It generated the heaviest and longest larvae at 65.03 mg and 18.32 mm, respectively, as well as pupae weighing 107.55 mg and 19.94 mm, respectively, and adults weighing 87.52 mg and 20.26 mm, respectively. It was also determined that fungi (C. indica) and ospor (B. clausii) promoted faster larval development than yeast (S. cerevisiae) and imutec (L. rhamnosus). Larval mortality was also greater in the imutec (L. rhamnosus) and yeast (S. cerevisiae) diets than the others. No pupal mortality was recorded in all diets. Furthermore, the protein content of Tenebrio. molitor raised on a diet including fungi (C. indica) was the highest at (375 g), with a content of 68.31%, followed by a concentration of (250 g) with a content of 67.84%, and wheat bran (1 kg) (normal diet) with the lowest content at 58.91%. T. molitor larvae fed a diet supplemented with bacterial and fungal had lower fat and ash content than bran-fed T. molitor larvae (standard diet). Wheat bran (normal diet) had the highest fat at 16.11%, and ash at 7.71%. Hence, it is concluded that wheat bran alone or diet containing fungi (C. indica) and ospor (B. clausii) performed better in terms of growth, and these diets and protein content are recommended for the mass rearing of mealworms. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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22 pages, 7072 KiB

Open AccessArticle

Improvements in Visual Aspects and Chemical, Techno-Functional and Rheological Characteristics of Cricket Powder (Gryllus bimaculatus) by Solvent Treatment for Food Utilization

by Barbara Catalano Damasceno, Mitsutoshi Nakajima, Noamane Taarji, Isao Kobayashi, Sosaku Ichikawa and Marcos A. Neves

Foods 2023, 12(7), 1422; https://doi.org/10.3390/foods12071422 - 27 Mar 2023

Cited by 3 | Viewed by 2942

Abstract

This study aimed to improve the visual aspects and chemical, techno-functional and rheological characteristics of Gryllus bimaculatus cricket powder through the use of different solvents, with the objective of using it as a protein source in food production. Four treatments (pH 5 aqueous solution, ethanol 20%, ethanol 99.5%, and hexane) were applied to the powder, and analyses were conducted to assess changes in the previously mentioned parameters. The results showed that the treatments led to an increase in protein concentration (from 55.4 to 72.5%) and a decrease in fat concentration (from 33.0 to 6.8%) in ethanol 99.5% treated powder, as well as a reduction in anti-nutritional compounds concentration, such as tannins (from 13.3 to 5.9 g/kg), in pH 5 treated powder, which is important for the nutritional value of the final product. The color of the powders was improved, being lighter after hexane and ethanol 99.5% treatments due to the removal of melanin with the defatting process. Flowability, water, and oil holding capacity were also improved in the defatted powders. All the results suggest that the main composition of the powder directly influences the analyzed parameters. These findings suggest that cricket powder treated with solvents can be used as a protein source in different food applications. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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19 pages, 2746 KiB

Open AccessArticle

Characterization of Tenebrio molitor Larvae Protein Preparations Obtained by Different Extraction Approaches

by Alkmini-Anna Gkinali, Anthia Matsakidou and Adamantini Paraskevopoulou

Foods 2022, 11(23), 3852; https://doi.org/10.3390/foods11233852 - 29 Nov 2022

Cited by 15 | Viewed by 3689

Abstract

Edible insects have recently attracted research attention due to their nutritional value and low environmental footprint. Tenebrio molitor larva was the first insect species to be classified by European Food Safety Authority (EFSA) as safe for human consumption. However, it is thought that the incorporation of edible insect as an ingredient in a food product would be more appealing to consumers than being visible. The aim of the present study was to determine the physicochemical properties of the larvae meal and protein concentrates. Different methods to extract and recover proteins from defatted (DF) Tenebrio molitor larvae were applied; i.e., alkaline extraction (DF-ASP); isoelectric precipitation after alkaline extraction (DF-AIP); and NaCl treatment (DF-SSP), and the obtained protein fractions were characterized. The DF-ASP exhibited the highest protein extraction/recovery efficiency (>60%), while it was the most effective in decreasing the interfacial tension at the oil/water (o/w) interface. The DF-AIP had the highest protein content (75.1%) and absolute values of ζ-potential and the best ability to retain water (10.54 g/g) and stabilize emulsions at pH 3.0. The DF-SSP protein preparation had the highest oil binding capacity (8.62%) and solubility (~88%) at acidic pHs and the highest emulsifying activity (~86 m²/g). Electrophoresis of the protein preparations revealed proteins with different molecular weights, while the protein secondary structure was dominated by β-structures and α-helix. Protein concentrates with different properties were able to be recovered from Tenebrio molitor larvae, that could affect their interactions with other food ingredients and their behavior during processing or storage. These findings would be valuable guidance for the technological exploitation of larvae protein preparations in the development of food formulations. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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11 pages, 1107 KiB

Open AccessArticle

Substituting Whole Wheat Flour with Pigeon Pea (Cajanus cajan) Flour in Chapati: Effect on Nutritional Characteristics, Color Profiles, and In Vitro Starch and Protein Digestion

by Sirin Sachanarula, Praew Chantarasinlapin and Sirichai Adisakwattana

Foods 2022, 11(20), 3157; https://doi.org/10.3390/foods11203157 - 11 Oct 2022

Cited by 3 | Viewed by 3768

Abstract

Pigeon pea (Cajanus cajan (L.) Millsp.), a potential legume as an economical source of protein, is commonly cultivated in tropical and subtropical regions of the world. Therefore, pigeon pea may be potentially used as a substitute to improve the nutritional profile of foods. In the present study, the effect of substitution of whole wheat flour (WWF) with 20% and 40% pigeon pea flour (PPF) on the nutritional properties, color profiles, and starch and protein digestibility of chapati was investigated. The results showed that PPF had higher protein content but less carbohydrate than WWF. The protein content of chapati substituted with 20% and 40% PPF increased by 1.18 and 1.34 times, respectively, compared to WWF chapati, along with a marked decrease in carbohydrate content. Analyses further revealed an increase in the lightness and yellowness and a decrease in the redness of the chapati. Furthermore, glucose release from chapati with 20% and 40% PPF under simulated digestion was attenuated, corresponding to decreased hydrolysis and a predicted glycemic index. In the 40% PPF chapati, a significant reduction in slowly digestible starch (SDS) with increased resistant starch (RS) proportions was achieved without altering the effect on rapidly digestible starch (RDS). In addition, the level of amino-group residues was markedly elevated in 20% and 40% PPF substituted chapati compared to WWF chapati. These findings suggest that PPF can serve as a promising plant-based alternative ingredient to improve the nutrient value of chapati by reducing starch and increasing protein digestibility. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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13 pages, 7569 KiB

Open AccessArticle

Assessing Edible Filamentous Fungal Carriers as Cell Supports for Growth of Yeast and Cultivated Meat

by Minami Ogawa, Jaime Moreno García, Nitin Nitin, Keith Baar and David E. Block

Foods 2022, 11(19), 3142; https://doi.org/10.3390/foods11193142 - 9 Oct 2022

Cited by 10 | Viewed by 4077

Abstract

The growth and activity of adherent cells can be enabled or enhanced through attachment to a solid surface. For food and beverage production processes, these solid supports should be food-grade, low-cost, and biocompatible with the cell of interest. Solid supports that are edible can be a part of the final product, thus simplifying downstream operations in the production of fermented beverages and lab grown meat. We provide proof of concept that edible filamentous fungal pellets can function as a solid support by assessing the attachment and growth of two model cell types: yeast, and myoblast cells. The filamentous fungus Aspergillus oryzae was cultured to produce pellets with 0.9 mm diameter. These fugal pellets were inactivated by heat or chemical methods and characterized physicochemically. Chemically inactivated pellets had the lowest dry mass and were the most hydrophobic. Scanning electron microscope images showed that both yeast and myoblast cells naturally adhered to the fungal pellets. Over 48 h of incubation, immobilized yeast increased five-fold on active pellets and six-fold on heat-inactivated pellets. Myoblast cells proliferated best on heat-treated pellets, where viable cell activity increased almost two-fold, whereas on chemically inactivated pellets myoblasts did not increase in the cell mass. These results support the use of filamentous fungi as a novel cell immobilization biomaterial for food technology applications. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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15 pages, 2410 KiB

Open AccessArticle

Effect of High Hydrostatic Pressure Intensity on Structural Modifications in Mealworm (Tenebrio molitor) Proteins

by Abir Boukil, Alice Marciniak, Samir Mezdour, Yves Pouliot and Alain Doyen

Foods 2022, 11(7), 956; https://doi.org/10.3390/foods11070956 - 25 Mar 2022

Cited by 19 | Viewed by 3512

Abstract

Processing edible insects into protein extracts may improve consumer acceptability. However, a better understanding of the effects of food processing on the proteins is needed to facilitate their incorporation into food matrices. In this study, soluble proteins from Tenebrio molitor (10% w/v) were pressurized using high hydrostatic pressure (HHP) at 70–600 MPa for 5 min and compared to a non-pressurized control (0.1 MPa). Protein structural modifications were evaluated using turbidity measurement, particle-size distribution, intrinsic fluorescence, surface hydrophobicity, gel electrophoresis coupled with mass spectrometry, and transmission electron microscopy (TEM). The observed decrease in fluorescence intensity, shift in the maximum emission wavelength, and increase in surface hydrophobicity reflected the unfolding of mealworm proteins. The formation of large protein aggregates consisting mainly of hexamerin 2 and ⍺-amylase were confirmed by protein profiles on gel electrophoresis, dynamic light scattering, and TEM analysis. The typical aggregate shape and network observed by TEM after pressurization indicated the potential involvement of myosin and actin in aggregate formation, and these were detected by mass spectrometry. For the first time, the identification of mealworm proteins involved in protein aggregation phenomena under HHP was documented. This work is the first step in understanding the mealworm protein–protein interactions necessary for the development of innovative insect-based ingredients in food formulations. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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Review

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27 pages, 5153 KiB

Open AccessReview

Plant-Based Meat Analogues from Alternative Protein: A Systematic Literature Review

by Izalin Zahari, Karolina Östbring, Jeanette K. Purhagen and Marilyn Rayner

Foods 2022, 11(18), 2870; https://doi.org/10.3390/foods11182870 - 16 Sep 2022

Cited by 57 | Viewed by 15795

Abstract

This study aimed to conduct a systematic literature review (SLR) of the research performed in the plant-based meat analogues area. Historical, current, and future tendencies are discussed. The paper offers a comprehensive SLR coupled with a bibliometric analysis of the publication from 1972 to January 2022. The articles were obtained using a research string and precise inclusion and exclusion criteria from two prominent databases, Scopus and Web of Science (WoS). The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) flow technique was used to describe the data screening and selection. In total, 84 publications were selected for further analysis after a thorough literature assessment. From this study, six main themes were identified: (1) objectives of the study; (2) type of plant protein; (3) product type; (4) added ingredients; (5) texturization technique; and (6) quality assessment considered in the studies. Recent trends in publication imply that meat analogue technology is gaining prominence. This review revealed significant research on improving meat analogues via texturization. Even though extrusion is used industrially, the technique is still in its infancy and needs improvement. Future studies should focus more on fiber and protein–protein interactions, macromolecule conformation and mechanisms, diversifying or improving current methods, sensory attributes, and gastrointestinal absorption rate of each novel protein ingredient. Full article

(This article belongs to the Special Issue Food Proteins from Alternative Sources: Processing, Functionality, Properties, and Applications)

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