Narrative Review of the Current and Future Perspectives of Phycobiliproteins' Applications in the Food Industry: From Natural Colors to Alternative Proteins.

Vivid-colored phycobiliproteins (PBPs) have emerging potential as food colors and alternative proteins in the food industry. However, enhancing their application potential requires increasing stability, cost-effective purification processes, and consumer acceptance. This narrative review aimed to highlight information regarding the critical aspects of PBP research that is needed to improve their food industry potential, such as stability, food fortification, development of new PBP-based food products, and cost-effective production. The main results of the literature review show that polysaccharide and protein-based encapsulations significantly improve PBPs' stability. Additionally, while many studies have investigated the ability of PBPs to enhance the techno-functional properties, like viscosity, emulsifying and stabilizing activity, texture, rheology, etc., of widely used food products, highly concentrated PBP food products are still rare. Therefore, much effort should be invested in improving the stability, yield, and sensory characteristics of the PBP-fortified food due to the resulting unpleasant sensory characteristics. Considering that most studies focus on the C-phycocyanin from Spirulina, future studies should concentrate on less explored PBPs from red macroalgae due to their much higher production potential, a critical factor for positioning PBPs as alternative proteins.

Recent advances in engineering microorganisms for the production of natural food colorants.

Food colorants are frequently added to processed foods since color is an important tool in the marketing of food products, influencing consumer perceptions, preferences, and purchasing behavior. While synthetic dyes currently dominate the food colorant market, consumer concern regarding their safety and sustainability is driving a demand for their replacement with naturally derived alternatives. However, natural colorants are costly compared to their synthetic counterparts as the pigment content in the native sources is usually very low and extraction can be challenging. Recent advances in the engineering of microbial metabolism have sparked interest in the development of cell factories capable of producing natural colorants from renewable resources. This review summarizes major developments within metabolic engineering for the production of nature-identical food colorants by fermentation. Additionally, it highlights common applications, formulations, and physicochemical characteristics of prevalent pigment classes. Lastly, it outlines a workflow for accelerating the optimization of cell factories for the production or derivatization of nature-identical food colorants.

Anthocyanins as Natural Food Colorings: The Chemistry Behind and Challenges Still Ahead.

Anthocyanins are polyphenolic O-glycosides widely responsible for the bright red, purple, and blue colors in the plant kingdom, including a great variety of fruits and vegetables. Hence, they have attracted considerable scientific and industrial interest as potential natural food colorings. However, individual anthocyanins are intrinsically reactive molecules combining electrophilic, nucleophilic, and electron-donating properties. This reactivity may be not only a source of color diversity with, for instance, the formation of new pigments upon winemaking and storage but also a cause of great color instability involving a combination of reversible and irreversible mechanisms (e.g., water addition, autoxidation) leading to colorless products. Hence, using anthocyanin-rich plant extracts as food colorings requires a deep understanding of these color-damaging mechanisms and, no less importantly, of the color-stabilizing mechanisms developed by plants, including π-stacking interactions (self-association, copigmentation), metal binding, and a combination of both. The potential of anthocyanins from deeply colored vegetables, typically acylated by hydroxycinnamic acid residues, will be emphasized in that respect. Moreover, food-grade biopolymers (proteins, polysaccharides) may provide suitable matrices for ready-to-use formulations of anthocyanins as food colorings. In this short review, the mechanisms of color loss and color stabilization are discussed as a function of anthocyanin structure and environment, and some challenges still ahead are outlined.

Electrochemical detection of 4(5)-methylimidazole in aqueous solutions.

4(5)-methylimidazole (4-MeI) is a potential carcinogen widely used in food colours. EU regulations specify a maximum allowable concentration of 200 ppm for 4-MeI in caramel colours. This study reports an electrochemical determination technique for 4-MeI in caramel colours for the first time. The effect of pH and interference from air were studied to optimize the detection conditions on a glassy carbon electrode in aqueous alkaline solutions using square wave voltammetry (SWV) technique. The concentration of 4-MeI was quantitatively measured down to 10 µM (∼0.8 ppm). Traditional methods such as HPLC, GC, spectrometry and immunoassays involve either expensive instrumentation and reagents or time consuming preparation and detection processes. This study demonstrates the possibility of rapid and simple electrochemical determination of (4-MeI) in food colours with minimum workup using a portable potentiostat.

Removal of food dyes using biological materials via adsorption: A review.

It is alarming that synthetic food dyes (FD) are widely used in various industries and that these facilities discharge their wastewater into the environment without treating it. FDs mixed into industrial wastewater pose a threat to the environment and human health. Therefore, removing FDs from wastewater is very important. This review explores the burgeoning field of FD removal from wastewater through adsorption using biological materials (BMs). By synthesizing a wealth of research findings, this comprehensive review elucidates the diverse array of BMs employed, ranging from algae and fungi to agricultural residues and microbial biomass. Furthermore, this review investigates challenges in practical applications, such as process optimization and scalability, offering insights into bridging the gap between laboratory successes and real-world implementations. Harnessing the remarkable adsorptive potential of BMs, this review presents a roadmap toward transformative solutions for FD removal, promising cleaner and safer production practices in the food and beverage industry.

Stability mechanism of Monascus pigment-soy protein isolate-maltodextrin complex.

BACKGROUND: Monascus pigment (MP) is a natural food coloring with vital physiological functions but prone to degradation and color fading under light conditions. RESULTS: This study investigated the effect of complex formation of soybean protein isolate (SPI), maltodextrin (MD), and MP on the photostability of MP. Light stability was assessed through retention rate and color difference. Fluorescence spectroscopy (FS), circular dichroism (CD), Fourier-transform infrared (FTIR) spectroscopy, and X-ray diffraction (XRD) explored MP, SPI, and MD interactions, clarifying the MP-SPI-MD complex mechanism on the light stability of MP. Microstructure and differential scanning calorimetry (DSC) analyzed the morphology and thermal properties. The retention rate of MP increased to approximately 80%, and minimal color difference was observed when adding SPI and MD simultaneously. FS revealed hydrophobic interaction between MP and SPI. FTIR analysis showed intensity changes and peak shifts in amide I band and amide II band, which proved the hydrophobic interaction. CD showed a decrease in α-helix content and an increase in ß-sheet content after complex formation, indicating strengthened hydrogen bonding interactions. Scanning electron microscopy (SEM) analysis demonstrated that MP was attached to the surface and interior of complexes. XRD showed MP as crystalline, while SPI and MD were amorphous, complexes exhibited weakened or absent peaks, suggesting MP encapsulation. The results of DSC were consistent with XRD. CONCLUSION: SPI and MD enveloped MP through hydrogen bonding and hydrophobic interaction, ultimately enhancing its light stability and providing insights for pigment-protein-polysaccharide interactions and improving pigment stability in the food industry. © 2024 Society of Chemical Industry.

pH-dependent stability of major betalains in the encapsulated beetroot extracts (Beta vulgaris L.).

Betalain is a water-soluble pigment contained in Caryophyllales plants. It not only holds potential as a natural food colorant but also offers various health benefits, acting as an antioxidant. This study focused on analyzing the pH-dependent stability of encapsulated betalain pigments extracted from red beetroot (Beta vulgaris L.) using methods such as absorption spectroscopy, HPLC, and LC-MS. The major pigments identified were vulgaxanthin I, betanin, isobetanin, and neobetanin, alongside minor components, including three betaxanthin species and a degradation product known as betalamic acid. Spectrophotometric analyses revealed that above pH 8, the betalain peak at 435 nm decreased and red-shifted to a peak at 549 nm, a shift that could be reversed through neutral pH treatment. At pH 11, a new broad peak appeared at 410 nm and was identified as betalamic acid. To assess the pH-dependency of each betalain, the targeted betalains were separated and quantified through HPLC after incubation across a wide pH range of 2-11 and during storage. After 3 days of storage in highly alkaline conditions (pH 10-11), major betalains, with the exception of neobetanin, underwent significant degradation. Conversely, these pigments displayed relative stability in acidic conditions. In contrast, neobetanin showed vulnerability to acidic conditions but exhibited tolerance to alkaline pH levels of 10-11. The degradation product, betalamic acid, demonstrated a similar susceptibility to alkaline pH as betanins. In conclusion, the significant stability decrease under highly alkaline conditions results not only from the hydrolytic reaction of betalains but also from the degradation of betalamic acid itself. PRACTICAL APPLICATION: Encapsulation methods are used to enhance the stability of betalains against temperature variations; however, the effects of pH, especially when considering individual betalain species, are not well understood. Despite betalains exhibiting similar features and being suitable for a wide pH range from acid to alkaline conditions, they are significantly affected by alkaline pH levels exceeding 10, as well as by storage duration. This study demonstrated the application of encapsulation to pH-dependent stability, and the findings offer valuable insights and a fresh perspective on betalains as red biocolorants, extending their potential application to a wide range of pH-controlled food products.

Natural blue.

A food dye from a South American fruit has become a test case for the ethical development of natural resources.

Analysis of Chlorophylls/Chlorophyllins in Food Products Using HPLC and HPLC-MS Methods.

Of the different quality parameters of any food commodity or beverage, color is the most important, attractive and choice-affecting sensory factor to consumers and customers. Nowadays, food industries are interested in making the appearance of their food products attractive and interesting in order to appeal to consumers/customers. Natural green colorants have been accepted universally due to their natural appeal as well as their nontoxic nature to consumers. In addition, several food safety issues mean that natural green colorants are preferable to synthetic food colorants, which are mostly unsafe to the consumers but are less costly, more stable, and create more attractive color hues in food processing. Natural colorants are prone to degradation into numerous fragments during food processing, and thereafter, in storage. Although different hyphenated techniques (especially high-performance liquid chromatography (HPLC), LC-MS/HRMS, and LC/MS-MS are extensively used to characterize all these degradants and fragments, some of them are not responsive to any of these techniques, and some substituents in the tetrapyrrole skeleton are insensitive to these characterization tools. Such circumstances warrant an alternative tool to characterize them accurately for risk assessment and legislation purposes. This review summarizes the different degradants of chlorophylls and chlorophyllins under different conditions, their separation and identification using various hyphenated techniques, national legislation regarding them, and the challenges involved in their analysis. Finally, this review proposes that a non-targeted analysis method that combines HPLC and HR-MS assisted by powerful software tools and a large database could be an effective tool to analyze all possible chlorophyll and chlorophyllin-based colorants and degradants in food products in the future.

Effects of two food colorants on catalase and trypsin: Binding evidences from experimental and computational analysis.

Recently, growing concern has been paid to the toxicity of additives in food. The present study investigated the interaction of two commonly used food colorants, quinoline yellow (QY) and sunset yellow (SY), with catalase and trypsin under physiological conditions by fluorescence, isothermal titration calorimetry (ITC), ultraviolet-vis absorption, synchronous fluorescence techniques as well as molecular docking. Based on the fluorescence spectra and ITC data, both QY and SY could significantly quench the intrinsic fluorescence of catalase or trypsin spontaneously to form a moderate complex driven by different forces. Additionally, the thermodynamics results demonstrated QY bind more tightly to both catalase and trypsin than SY, suggesting QY poses more of a threat to two enzymes than SY. Furthermore, the binding of two colorants could not only lead to the conformational and microenvironmental alterations of both catalase and trypsin, but also inhibit the activity of two enzymes. This study provides an important reference for understanding the biological transportation of synthetic food colorants in vivo, and enhancing their risk assessment on food safety.

Comprehensive chlorophyll composition of commercial green food colorants and coloring foodstuffs by HPLC-ESI-QTOF-MS/MS: Chlorophyllins.

Consumers demand higher levels of food quality and safety, so food legislative organizations need full knowledge of food composition to develop regulations that guarantee quality and safety criteria. This is the context for green natural food colorants and the new category green "coloring foodstuffs". We have exploited the capabilities of targeted metabolomics assisted by powerful software and algorithms to unravel the comprehensive chlorophyll composition in commercial samples of both colorant categories. With the aid of an in-house library, at first, seven new chlorophylls have been identified, among all the samples analyzed, providing data on their structural configuration. Next, taking advantage of an expert-curated database, eight more chlorophylls non-described previously have been found, which will be significant for the chemistry of chlorophylls. Finally, we have deciphered the sequence of chemical reactions that take place during the manufacturing of green food colorants and propose the whole pathway that explains the occurrence of the chlorophylls they contain.

Recent advancements in natural colorants and their application as coloring in food and in intelligent food packaging.

Colorants are widely employed in the food industry as an essential ingredient in many products since color is one of the most valued attributes by consumers. Furthermore, the utilization of colorants is currently being extended to the food packaging technologies. The objective of this review was to compile recent information about the main families of natural coloring compounds, and to describe their real implications in food coloring. In addition, their technological use in different food systems (namely, bakery products, beverages, meat and meat products, and dairy products) and their utilization in intelligent packaging to monitor the freshness of foodstuffs with the aim of extending food shelf life and improving food properties was discussed. The potential of using natural colorant in different food to improve their color has been demonstrated, although color stability is still a challenging task. More interestingly, the application of intelligent colorimetric indicators to exhibit color changes with variations in pH can enable real-time monitoring of food quality.

Characterisation of iron oxide-containing pearlescent pigments used as food colourants: nano-labelling required in the EU?

Pearlescent pigments are used as colourants to increase the attractiveness of food products, especially in the patisserie and confectionery sector. They can be seen as composite materials and consist of thin potassium aluminium silicate (E 555, mica) platelets as carrier material, coated with a thin metal oxide layer of TiO2 (E 171) and/or iron oxides (E 172). The European Food Safety Authority stated in 2020 that mica-based pearlescent pigments as a whole should be evaluated as new food additives. Obtaining dependable data for particle size and layer thickness of these pigments is crucial both for the demanded food additive evaluation itself and also for the nanomaterial labelling assessment of products containing these food colourants according to the 'Food Information to Consumers' regulation. Since it was found in a previous study on TiO2-containing pearlescent pigments (silver and golden coloured) that the coating consisted of nanoscaled constituent titanium oxide particles, in this follow-up study we investigated whether Fe2O3-containing pearlescent pigments exhibit a similar nanostructured morphology. For this purpose, five commercially-available food products containing these pigments were investigated. Static light scattering and flow particle image analysis were used as screening methods to determine the mica platelet size. Scanning electron microscopy combined with energy-dispersive X-ray spectroscopy was used for nanostructure analysis of the metal oxide coating. The carrier mica platelets were 34-96 µm in diameter and 300-800 nm thick. The coating thickness was found to be in the range of 75-105 nm, with the constituent round shaped iron oxide particles contained therein having a minimum Feret diameter of 37-64 nm.

Ascomycota as a source of natural colorants.

In the last few decades, there has been a great demand for natural colorants. Synthetic colorants are known to be easy to produce, are less expensive, and remain stable when subjected to chemical and physical factors. In addition, only small amounts are required to color any material, and unwanted flavors and aromas are not incorporated into the product. Natural colorants present in food, in addition to providing color, also have biological properties and effects that aid in the prevention and cure of many diseases. The main classes of colorants produced by phylum Ascomycota include polyketides and carotenoids. A promising producer of colorants should be able to assimilate a variety of sources of carbon and nitrogen and also exhibit relative stability. The strain should not be pathogenic, and its product should not be toxic. Production processes should also provide the expected color with a good yield through simple extraction methods. Research that seeks new sources of these compounds should continue to seek products of biotechnological origin in order to be competitive with products of synthetic and plant origin. In this review, we will focus on the recent studies on the main producing species, classes, and metabolic pathways of colorants produced by this phylum, historical background, impact of synthetic colorants on human health and the environment, social demand for natural colorants and also an in-depth approach to bioprocesses (influences on production, optimization of bioprocess, extraction, and identification), and limitations and perspectives for the use of fungal-based dyes.

Insight into the Progress on Natural Dyes: Sources, Structural Features, Health Effects, Challenges, and Potential.

(1) Background: Dyes play an important role in food, medicine, textile, and other industries, which make human life more colorful. With the increasing demand for food safety, the development of natural dyes becomes more and more attractive. (2) Methods: The literature was searched using the electronic databases PubMed, Web of Science, and SciFinder and this scoping review was carried out following Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). (3) Results: 248 articles were included in this review. This review summarizes the research progress on natural dyes in the last ten years. According to structural features, natural dyes mainly include carotenoids, polyphenols, porphyrins, and alkaloids, and some of the newest dyes are summarized. Some pharmacological activities of carotenoids, anthocyanin, curcumin, and betalains in the last 10 years are summarized, and the biological effects of dyes regarding illumination conditions. The disadvantages of natural dyes, including sources, cost, stability, and poor bioavailability, limit their application. Here, some feasible strategies (potential resources, biotechnology, new extraction and separation strategies, strategies for improving stability) are described, which will contribute to the development and utilization of natural dyes. (4) Conclusion: Natural dyes show health benefits and potential in food additives. However, it is necessary for natural dyes to pass toxicity tests and quality tests and receive many regulatory approvals before their final entry into the market as food colorants or as drugs.

Stability of natural food colorants derived from onion leaf wastes.

Plant anthocyanins have widely been employed as natural food colorants. However, their instability restricts many of their applications in food industry. In this study, anthocyanins were extracted from onion outer scales, using aqueous solutions with or without added cyclodextrins (CDs). The results indicated that when cyclodextrins were included in the extraction medium, the anthocyanins were stable or even had improved and augmented color intensity upon storage or following thermal and UV-light treatments over a broad pH range (2.0-7.0). FT-IR and UV-Vis spectroscopy measurements confirmed the formation of inclusion complexes between CDs and anthocyanins and the presence of pyranosyl groups (pyranoanthocyanins) upon heating. Overall, the stability of onion anthocyanins under various environmental stresses, often encountered during food processing and storage, indicates that the natural color extract from onion outer scales can be of value as an interesting colorant alternative for food applications.

Physicochemical properties, degradation kinetics, and antioxidant capacity of aqueous anthocyanin-based extracts from purple carrots compared to synthetic and natural food colorants.

As a means to evaluate the potential of carrot anthocyanins as food colorants and nutraceutical agents, we investigated the physicochemical stability and antioxidant capacity of purple carrot extracts under different pH (2.5-7.0) and temperature (4-40 °C) conditions, in comparison to a commercial synthetic (E131) and a natural grape-based (GRP) colorant. During incubation, the colorants were weekly-monitored for various color parameters, concentration of anthocyanins and phenolics, and antioxidant capacity. Carrot colorants were more stable than GRP; and their thermal stability was equal (at 4 °C) or higher than that of E131 (at 25-40 °C). Carrot anthocyanins had lower degradation rate at low pH and temperature, with acylated anthocyanins (AA) being significantly more stable than non-acylated anthocyanins (NAA). Anthocyanins acylated with feruloyl and coumaroyl glycosides were the most stable carrot pigments. The higher stability of carrot colorants is likely due to their richness in AA and -to a lesser extent- copigmentation with other phenolics.

The protective effect of natural phenolic compound on the functional and structural responses of inhibited catalase by a common azo food dye.

In this research retrieval effects of natural yellow (NY) on the performance of carmoisine (CAR) inhibited bovine liver catalase (BLC) was studied using multispectral and theoretical methods. Kinetic studies showed that CAR inhibited BLC through competitive inhibition (IC50 value of 2.24 × 10-6 M) while the addition of NY recover the activity of CAR-BLC up to 82% in comparison with the control enzyme. Circular dichroism data revealed that NY can repair the structural changes of BLC, affected by CAR. Furthermore, an equilibrium dialysis study indicated that NY could reduce the stability of the CAR-catalase complex. The surface plasmon resonance (SPR) data analysis indicated a high affinity of NY to BLC compared to CAR and the binding of NY led to a decrease in the affinity of the enzyme to the inhibitor. On the other hand, fluorescence and molecular docking studies showed that the quenching mechanism of BLC by CAR occurs through a static quenching process, and van der Waals forces and hydrogen bonding play a crucial role in the binding of CAR to BLC. MLSD data demonstrated that NY could increase the binding energy of CAR-BLC complex from -7.72 kJ mol-1 to -5.9 kJ mol-1, leading to complex instability and catalase activity salvage.

Recent advances in Ponceau dyes monitoring as food colorant substances by electrochemical sensors and developed procedures for their removal from real samples.

Ponceau dyes are one of the food coloring materials that are added to various pharmaceutical, health and food products and give them an appearance. These dyes contain contaminants such as Benzidine, 4-Aminobiphenyl, and 4-Aminoazobenzene that are safe in small amounts, but they are not approved by the US Food and Drug Administration (US-FDA) for human consumption. This study comprehensively was reviewed the properties, applications, chemistry, and toxicity of Ponceau dyes as food colorant substances. Electroanalysis of Ponceau dyes was discussed in detail, and the various electrochemical sensors used to detect and monitor these dyes as food colorant were examined. The applied methods of removing and degradation of these dyes in municipal and industrial wastes were also discussed. Conclusions and future perspectives to motivate future research were also explored.

Red Tomato Products as an Alternative to Reduce Synthetic Dyes in the Food Industry: A Review.

Most dyes used in the food industry are synthetic and can be a health hazard. Red tomato may serve as a natural alternative dye to replace synthetic colorants. This study aimed to review the literature on the addition of red tomato products (powder tomato, paste, freeze-dried, tomato peel powder, tomato pomace) to reduce the usage of synthetic dyes in the food industry. Red tomato products have been used as coloring in pasta, bologna, sausages, cookies, crackers, macaroons, hamburgers, breads, muffins, cheeses, and nuggets. The trans-cis isomerization of lycopene by oxidative processes directly affects the color of the pigment. The lycopene contained in tomato has antioxidant activity and could reduce or eliminate other oxidants and/or synthetic preservatives in food. Moreover, tomatoes in foods have high sensory scores, nutritional appeal, and marketing potential. However, its use as a food colorant has been not extensively explored. Therefore, further studies are still required, especially on the stability of carotenoids in tomatoes used in processed foods.