Biotechnological approaches for producing natural pigments in yeasts.

Pigments are widely used in the food, cosmetic, textile, pharmaceutical, and materials industries. Demand for natural pigments has been increasing due to concerns regarding potential health problems and environmental pollution from synthetic pigments. Microbial production of natural pigments is a promising alternative to chemical synthesis or extraction from natural sources. Here, we discuss yeasts as promising chassis for producing natural pigments with their advantageous traits such as genetic amenability, safety, rapid growth, metabolic diversity, and tolerance. Metabolic engineering strategies and optimizing strategies in downstream process to enhance production of natural pigments are thoroughly reviewed. We discuss the challenges, including expanding the range of natural pigments and improving their feasibility of industrial scale-up, as well as the potential strategies for future development.

Natural pigments in the food industry: Enhancing stability, nutritional benefits, and gut microbiome health.

Natural pigments are increasingly favored in the food industry for their vibrant colors, fewer side effects and potential health benefits compared to synthetic pigments. However, their application in food industry is hindered by their instability under harsh environmental conditions. This review evaluates current strategies aimed at enhancing the stability and bioactivity of natural pigments. Advanced physicochemical methods have shown promise in enhancing the stability of natural pigments, enabling their incorporation into food products to enhance sensory attributes, texture, and bioactive properties. Moreover, recent studies demonstrated that most natural pigments offer health benefits. Importantly, they have been found to positively influence gut microbiota, in particular their regulation of the beneficial and harmful flora of the gut microbiome, the reduction of ecological dysbiosis through changes in the composition of the gut microbiome, and the alleviation of systemic inflammation caused by a high-fat diet in mice, suggesting a beneficial role in dietary interventions.

A review on the role of pH-sensitive natural pigments in biopolymers based intelligent food packaging films.

As food packaging evolves, consumer interests are shifting from traditional to intelligent food packaging systems. Intelligent packaging includes active components that display changes in a visual or interactive form perceivable by consumers. This offers real-time monitoring of the quality and shelf life of the packaged food and enhances transparency. For example, pH-sensitive natural pigment-based films change color in response to variations in pH levels, enabling the film/labels to reflect alterations in the acidity or basicity of the food inside the package. Natural pigments like anthocyanins, curcumin, betalains, chlorophyll, and carotenoids have been comprehensively reported for developing biodegradable pH-sensitive films of starch, protein, chitosan, and cellulose. Natural pigments offer great compatibility with these biopolymers and improve the other performance parameters of the films. However, these films still lack the strength and versatility of petroleum-based synthetic plastic films. But these films can be used as an indicator and combined with primary packaging to monitor freshness, time-temperature, and leak for muscle foods, dairy products, fruits and vegetables, and bakery products. Therefore, this review provides a detailed overview of pH-sensitive pigments, their compatibility with natural polymers, their role in film performance in monitoring, and their food packaging applications.

A review on improving the sensitivity and color stability of naturally sourced pH-sensitive indicator films.

Naturally sourced pH-sensitive indicator films are of interest for real-time monitoring of food freshness through color changes because of their safety. Therefore, natural pigments for indicator films are required. However, pigment stability is affected by environmental factors, which can in turn affect the sensitivity and color stability of the pH-sensitive indicator film. First, natural pigments (anthocyanin, betalain, curcumin, alizarin, and shikonin) commonly used in pH-sensitive indicator films are presented. Subsequently, the mechanisms behind the change in pigment color under different pH environments and their applications in monitoring food freshness are also described. Third, influence factors, such as the sources, types, and pH sensitivity of pigments, as well as environmental parameters (light, temperature, humidity, and oxygen) of sensitivity and color stability, are analyzed. Finally, methods for improving the pH-sensitive indicator film are explored, encapsulation of natural pigments, incorporation of a hydrophobic film-forming matrix or function material, and protective layer have been shown to enhance the color stability of indicator films, the addition of copigments or mental ions, blending of different natural pigments, and the utilization of electrospinning have been proved to increase the color sensitivity of indicator films. This review could provide theoretical support for the development of naturally sourced pH-sensitive indicator films with high stability and sensitivity and facilitate the development in the field of monitoring food freshness.

Natural Pigments Recovery from Food By-Products: Health Benefits towards the Food Industry.

Given the health risks associated with synthetic colorants, natural pigments have emerged as a promising alternative. These renewable choices not only provide health benefits but also offer valuable technical and sensory properties to food systems. The effective application of natural colorants, however, requires the optimization of processing conditions, exploration of new sources, and development of novel formulations to ensure stability and maintain their inherent qualities. Several natural pigment sources have been explored to achieve the broad color range desired by consumers. The purpose of this review is to explore the current advances in the obtention and utilization of natural pigments derived from by-products, which possess health-enhancing properties and are extracted through environmentally friendly methods. Moreover, this review provides new insights into the extraction processes, applications, and bioactivities of different types of pigments.

Microencapsulated granaticins from Streptomyces vilmorinianum YP1: Optimization, physiochemical characterization and storage stability.

Granaticins are natural pigments derived from microorganisms with promising bioactivity. However, their practical applications have been restricted due to inherent instability. To improve the stability of granaticins from the novel strain Streptomyces vilmorinianum YP1, microcapsules were prepared using gum Arabic (GA) by a freeze-drying method. The optimal parameters for microencapsulation were determined using response surface methodology. Under the optimal conditions (GA 9.2% (v/v), a wall/-core ratio 4.8 (w/w), encapsulating temperature 29 °C), the maximum encapsulation efficiency achieved was 93.64%. The microcapsules were irregular single crystals with an average particle size of 206.37 ± 2.51 nm. Stability testing indicated improved stability of the microencapsulated granaticins. Notably, granaticnic B retention increased by 17.0% and 6.6% after exposure to sunlight and storage at 4 °C, respectively. These finding suggest that GA as a well material significantly enhances the stability of granaticins from S. vilmorinianum YP1, facilitating their potential applications.

Ultrasound-assisted extraction and encapsulation of betalain from prickly pear: Process optimization, in-vitro digestive stability, and development of functional gummies.

The study aimed to extract and encapsulate betalain pigment from prickly pear (Opuntia ficus-indica) using ultrasound-assisted extraction and eco-friendly glycerol. Subsequent analysis encompassed assessing its thermal stability, shelf-life, bio-accessibility, and biological properties. The process optimization employed Response Surface Methodology (RSM), focusing on glycerol concentration (20-50 %), sample to solvent ratio (1:10-1:20), temperature (30-60 °C), and time (10-30 min). Optimal conditions were determined as 23.15 % glycerol, 1:10 sample to solvent ratio, 10.43 min treatment time, and 31.15 °C temperature. Under these conditions, betalain content reached 858.28 mg/L with a 93.76 % encapsulation efficiency. Thermal stability tests (80-180 °C; 30 & 60 min) showed degradation of betalain with higher temperatures and longer durations, affecting the visual aspect (ΔE) of the pigment. Encapsulated betalain exhibited favorable shelf stability, with optimal storage life of 404.27 days at 4 °C in amber conditions, compared to 271.99 days at 4 °C without amber, 141.92 days at 25 °C without amber, and 134.22 days at 25 °C with amber. Bio-accessibility of encapsulated betalain was significantly higher (2.05 ± 0.03 %) than conventionally extracted pigment (1.03 ± 0.09 %). The encapsulated pigment displayed strong anti-inflammatory properties in dosages of 2-20 µL, with no cytotoxic effects. Additionally, incorporation into gummies was successful and visually approved by sensory panellists. Glycerol proved to be a green encapsulating agent for betalain, offering high shelf life and bio-accessibility, making it suitable for food industry applications. The encapsulated pigment demonstrated robust thermal stability and shelf life, making it suitable for food industry applications. This study highlights glycerol's potential as a sustainable alternative for natural pigment extraction.

Carotenoids and Intestinal Harmony: Exploring the Link for Health.

Carotenoids, prominent lipid-soluble phytochemicals in the human diet, are responsible for vibrant colours in nature and play crucial roles in human health. While they are extensively studied for their antioxidant properties and contributions to vitamin A synthesis, their interactions with the intestinal microbiota (IM) remain poorly understood. In this study, beta (ß)-carotene, lutein, lycopene, a mixture of these three pigments, and the alga Osmundea pinnatifida were submitted to simulated gastrointestinal digestion (GID) and evaluated on human faecal samples. The results showed varying effects on IM metabolic dynamics, organic acid production, and microbial composition. Carotenoid exposure influenced glucose metabolism and induced the production of organic acids, notably succinic and acetic acids, compared with the control. Microbial composition analysis revealed shifts in phyla abundance, particularly increased Pseudomonadota. The α-diversity indices demonstrated higher diversity in ß-carotene and the pigments' mixture samples, while the ß-diversity analysis indicated significant dissimilarity between the control and the carotenoid sample groups. UPLC-qTOF MS analysis suggested dynamic changes in carotenoid compounds during simulated fermentation, with lutein exhibiting distinct mass ion fragmentation patterns. This comprehensive research enhances our understanding of carotenoid-IM interactions, shedding light on potential health implications and the need for tailored interventions for optimal outcomes.

Effects of fermentation on the structures of yellow compounds in citrus pomace.

To enhance the stability and light resistance of the yellow compounds in citrus pomace, our study successfully isolated and purified five compounds using ultrasonic-assisted extraction and column chromatography. The identified compounds include methyl linoleate, (2-ethyl)hexyl phthalate, 1,3-distearoyl-2-oleoylglycerol, 6,6-ditetradecyl-6,7-dihydroxazepin-2(3H)-one, and n-octadeca-17-enoic acid. The monomers extracted from fresh pomace, compounds 1 and 2, exhibit structural similarities to flavonoids and carotenoids. In contrast, the polymers isolated from fermented pomace, compounds 3, 4, and 5, share structural units with the fresh pomace compounds, indicating the transformation to stable polymeric forms. This suggests that the microbial fermentation process not only enhances the value of citrus pomace, but also provides a promising pathway for the synthesis of natural antioxidant yellow pigments with far-reaching theoretical and practical significance.

Alternative green solvents associated with ultrasound-assisted extraction: A green chemistry approach for the extraction of carotenoids and chlorophylls from microalgae.

This study proposes a method for the ultrasonic extraction of carotenoids and chlorophyll from Scenedesmus obliquus and Arthrospira platensis microalgae with green solvents. Ethanol and ethanolic solutions of ionic liquids were tested with a variety of extraction parameters, including number of extractions, time of extraction, and solid-liquid ratio R(S/L), to determine the optimal conditions. After selecting the most effective green solvent (ethanol), the process conditions were established: R(S/L) of 1:10, three extraction cycles at 3 min each), giving an extraction yield of 2602.36 and 764.21 µgcarotenoids.gdried biomass-1; and 22.01 and 5.81 mgchlorophyll.gdried biomass-1 in S. obliquus and A. platensis, respectively. The carotenoid and chlorophyll extracts obtained using ethanol were shown to be potent scavengers of peroxyl radical, being 5.94 to 26.08 times more potent α-tocopherol. These findings pave the way for a green strategy for valorizing microalgal biocompounds through efficient and environmentally friendly technological processes.

Enhancing phycocyanin yield from Spirulina sp. under salt stress using various extraction methods.

Phycocyanin, a blue-coloured pigment, predominantly found and derived from Spirulina sp., has gained researchers' interest due to its vibrant hues and other attractive properties like antioxidant and anti-microbial. However, the lack of reliable and sustainable phycocyanin extraction strategies without compromising the quality has hindered the scaling up of its production processes for commercial purposes. Here in this study, phycocyanin was extracted from wet and dry biomass Spirulina sp., using three different physical cell disruption methods (ultrasonication, homogenization, and freeze-thaw cycles) combined with two different buffers (phosphate buffer and acetate buffer) and water (as control). The result showed that the freeze-thaw method combined with acetate buffer produced the highest yield (25.013 ± 2.572 mg/100 mg) with a purity ratio of 0.806 ± 0.079. Furthermore, when subjected to 30% w/v salt stress, 1.9 times higher phycocyanin yield with a purity ratio of 1.402 ± 0.609 was achieved using the previously optimized extraction method.

Ultrasound-assisted extraction and concentration of phenolic compounds from jabuticaba sabará (Plinia peruviana (Poir.) Govaerts) peel by nanofiltration membrane.

Jabuticaba peel, rich in antioxidants, offering health benefits. In this study, the extraction of phenolic compounds from jabuticaba peel using ultrasound-assisted (UA) and their subsequent concentration by nanofiltration (NF) employing a polyamide 200 Da membrane was evaluated. The UA extractions were conducted using the Central Composite Rotatable Design (CCRD) 22 methodology, with independent variables extraction time (11.55 to 138 min) and temperature (16.87 to 53.3 °C), and fixed variables mass to ethanol solution concentration at pH 1.0 (1:25 g/mL), granulometry (1 mm), and ultrasonic power (52.8 W). The maximum concentrations obtained were 700.94 mg CE/100 g for anthocyanins, 945.21 mg QE/100 g for flavonoids, 133.19 mg GAE/g for phenols, and an antioxidant activity IC50 of 24.36 µg/mL. Key phenolic compounds identified included cyanidin-3-glucoside, delphinidin-3-glucoside, and various acids like syringic and gallic. NF successfully concentrated these compounds, enhancing their yield by up to 45%. UA and NF integrate for sustainable extraction.

Nonlinearity and anthocyanin colour expression: A mathematical analysis of anthocyanin association kinetics and equilibria.

Anthocyanins are polyphenolic compounds that provide pigmentation in plants as reflected by pH-dependent structural transformations between the red flavylium cation, purple quinonoidal base, blue quinonoidal anion, colourless hemiketal, and pale yellow chalcone species. Thermodynamically stable conditions of hydrated plant cell vacuoles in vivo correspond to the colourless hemiketal, yet anthocyanin colour expression appears in an important variety of hues within plant organs such as flowers and fruit. Moreover, anthocyanin colour from grape berries is significant in red winemaking processes as it plays a crucial role in determining red wine quality. Here, nonlinear ordinary differential equations were developed to represent the evolution in concentration of various anthocyanin species in both monomeric (chemically reactive) and self-associated (temporally stable) forms for the first time, and simulations were verified experimentally. Results indicated that under hydrating conditions, anthocyanin pigmentation is preserved by self-association interactions, based on pigmented monomeric anthocyanins experiencing colour loss whereas colour-stable self-associated anthocyanins increase in concentration nonlinearly over time. In particular, self-association of the flavylium cation and the quinonoidal base was shown to influence colour expression and stability within Geranium sylvaticum flower petals and Vitis vinifera grape skins. This study ultimately characterises fundamental mechanisms of anthocyanin stabilisation and generates a quantitative framework for anthocyanin-containing systems.

An Update in Computational Methods for Environmental Monitoring: Theoretical Evaluation of the Molecular and Electronic Structures of Natural Pigment-Metal Complexes.

Metals are beneficial to life, but the presence of these elements in excessive amounts can harm both organisms and the environment; therefore, detecting the presence of metals is essential. Currently, metal detection methods employ powerful instrumental techniques that require a lot of time and money. Hence, the development of efficient and effective metal indicators is essential. Several synthetic metal detectors have been made, but due to their risk of harm, the use of natural pigments is considered a potential alternative. Experiments are needed for their development, but they are expensive and time-consuming. This review explores various computational methods and approaches that can be used to investigate metal-pigment interactions because choosing the right methods and approaches will affect the reliability of the results. The results show that quantum mechanical methods (ab initio, density functional theory, and semiempirical approaches) and molecular dynamics simulations have been used. Among the available methods, the density functional theory approach with the B3LYP functional and the LANL2DZ ECP and basis set is the most promising combination due to its good accuracy and cost-effectiveness. Various experimental studies were also in good agreement with the results of computational methods. However, deeper analysis still needs to be carried out to find the best combination of functions and basis sets.

Antarctic fungi produce pigment with antimicrobial and antiparasitic activities.

Natural pigments have received special attention from the market and industry as they could overcome the harm to health and the environmental issues caused by synthetic pigments. These pigments are commonly extracted from a wide range of organisms, and when added to products they can alter/add new physical-chemical or biological properties to them. Fungi from extreme environments showed to be a promising source in the search for biomolecules with antimicrobial and antiparasitic potential. This study aimed to isolate fungi from Antarctic soils and screen them for pigment production with antimicrobial and antiparasitic potential, together with other previously isolated strains A total of 52 fungi were isolated from soils in front of the Collins Glacier (Southeast border). Also, 106 filamentous fungi previously isolated from the Collins Glacier (West border) were screened for extracellular pigment production. Five strains were able to produce extracellular pigments and were identified by ITS sequencing as Talaromyces cnidii, Pseudogymnoascus shaanxiensis and Pseudogymnoascus sp. All Pseudogymnoascus spp. (SC04.P3, SC3.P3, SC122.P3 and ACF093) extracts were able to inhibit S. aureus ATCC6538 and two (SC12.P3, SC32.P3) presented activity against Leishmania (L.) infantum, Leishmania amazonensis and Trypanossoma cruzii. Extracts compounds characterization by UPLC-ESI-QToF analysis confirmed the presence of molecules with biological activity such as: Asterric acid, Violaceol, Mollicellin, Psegynamide A, Diorcinol, Thailandolide A. In conclusion, this work showed the potential of Antartic fungal strains from Collins Glacier for bioactive molecules production with activity against Gram positive bacteria and parasitic protozoas.

Microbial pigments: Sources, current status, future challenges in cosmetics and therapeutic applications.

Color serves as the initial attraction and offers a pleasing aspect. While synthetic colorants have been popular for many years, their adverse environmental and health effects cannot be overlooked. This necessitates the search for natural colorants, especially microbial colorants, which have proven and more effective. Pigment-producing microorganisms offer substantial benefits. Natural colors improve product marketability and bestow additional benefits, including antioxidant, antiaging, anticancer, antiviral, antimicrobial, and antitumor properties. This review covers the various types of microbial pigments, the methods to enhance their production, and their cosmetic and therapeutic applications. We also address the challenges faced during the commercial production of microbial pigments and propose potential solutions.

Extraction pathways and purification strategies towards carminic acid as natural-based food colorant: A comprehensive review.

As a current trend of fabricating healthier products, food manufacturing companies seek for natural-based food colorant aiming to replace the synthetic ones, which apart from meeting sensorial and organoleptic aspects, they can also act as health promoters offering additional added value. Carminic acid is a natural based food colorant typically found in several insect taxa. However, there are current approaches which pursue the production of this natural pigment via biotechnological synthesis. To date, this colorant has been intensively applied in the manufacture of several food items. Unfortunately, one of the main limitations deals with the establishment of the right protocol of extraction and purification of this component since there is no report analyzing the main extraction techniques for obtaining carminic acid. Therefore, this review, for the first time, comprehensively analyzes the ongoing strategies and protocols proposed by scientists towards either extraction or purification of carminic acid from its origin source, and from biotechnological systems. Emphasis has been focused on the main findings dealing with extraction techniques and the relevant insights in the field. A detailed discussion is provided on the advantages and drawbacks of the reported extraction and purification methods, main solvents used and their key interactions with target molecules.

Changing Despicable Me: Potential replacement of azo dye yellow tartrazine for pequi carotenoids employing ionic liquids as high-performance extractors.

Color is a crucial sensory attribute that guides consumer expectations. A high-performance pequi carotenoid extraction process was developed using ionic liquid-based ethanolic solutions and a factorial design strategy to search for a potential substitute for the artificial azo dye yellow tartrazine. All-trans-antheraxanthin was identified with HPLC-PAD-MSn for the first time in pequi samples. [BMIM][BF4] was the most efficient ionic liquid, and the maximization process condition was the solid-liquid ratio R(S/L) of 1:3, the co-solvent ratio R(IL/E) of 1:1 ([BMIM][BF4]: ethanol), and three cycles of extraction with 300 s each and yielded 107.90 µg carotenoids/g of dry matter. The ionic liquid-ethanolic solution recyclability was accomplished by freezing and precipitating with an average recovery of 79 %. In CIELAB parameters, pequi carotenoid extracted with [BMIM][BF4] was brighter and yellower than the artificial azo dye yellow tartrazine. A color change of 11.08 and a hue* difference of 1.26° were obtained. Furthermore, carotenoids extracted with [BMIM][BF4] showed antioxidant activity of 35.84 µmol of α-tocopherol. These findings suggest the potential of employing the pequi carotenoids to replace the artificial azo dye yellow tartrazine in foods for improved functional properties.

Natural pigments (anthocyanins and chlorophyll) and antioxidants profiling of European red and green gooseberry (Ribes uva-crispa L.) extracted using green techniques (UAE-citric acid-mediated extraction).

Green techniques to extract natural pigments are gaining prominence among consumers and food industries. This trend is predominantly due to the harmful effects imparted by commonly used synthetic dyes and the unwarranted stress created on our ecosystem. The objectives of this study were to obtain natural pigments (anthocyanins and chlorophyll) from Estonian-gown European green and red gooseberries by ultrasonic-assisted citric acid-mediated extraction method and perform antioxidant profiling (quantification via HPLC analysis). Green gooseberry extracts showed lower content of targeted compounds, with low concentrations of rutin (0.7-1.2 mg/L) and quercetin 3-glucoside (0.9-1.3 mg/L), while in the red gooseberry extracts, the amount was slightly higher (1.4-6.9 and 1.0-1.3 mg/L, respectively) with 0.6-6.8 mg/L cyanidin 3-glucoside and 0.32-0.35 mg/L peonidin 3 glucoside recorded. Further, the yield of anthocyanins ranged between 1.14-1.79 and 1.86-3.63 mg/100 g in green and red gooseberries, respectively. Total phenols ranged between 162-392 and 263-987 mg GAE/100 g in green and red gooseberry extracts, respectively. The DPPH free radicals scavenging activity showed 73-86% and 87-91% inhibition in both green and red gooseberry, respectively. Results showed significant improvements in pigment extraction with higher values obtained for targeted antioxidant compounds using conventional and UAE extraction (aqueous extract), thus confirming that green extractions are a reliable technique to obtain pigments of interest from natural sources. The results support consumers' demand and open up the avenue to explore pigments as natural colourants in food and cosmetics applications.

Biodiversity and biological applications of marine actinomycetes-Abu-Qir Bay, Mediterranean Sea, Egypt.

BACKGROUND: The ability of actinomycetes to produce bioactive secondary metabolites makes them one of the most important prokaryotes. Marine actinomycetes are one of the most important secondary metabolites producers used for pharmaceuticals and other different industries. RESULTS: In this study, the promising actinomycetes were isolated from Abu-Qir Bay. Four different media named as starch nitrate, starch casein, glycerol asparagine, and glycerol glycine were used as a preliminary experimental media to study the role of the medium components on the counts of actinomycetes in sediment samples. The results indicated that starch casein medium reported the highest counts (30-63 CFU/g) in all the tested sites. Lower counts were detected on starch nitrate and glycerol asparagine. On the other hand, glycerol glycine medium gave the lowest counts (15-48 CFU/g). Abu-Qir8 harbored the highest average count of actinomycetes (63 CFU/g), followed by Abu-Qir1 (48 CFU/g). The lower counts were detected in Abu-Qir5 and Abu-Qir7 (26 and 29 CFU/g, respectively). A total of 12 pure obtained actinomycetes isolates were subjected to morphological, physiological, and biochemical characterization. The selected actinobacterial isolates were subjected to numerical analysis, and the majority of isolates were grouped into four main clusters (A, B, C, & D), and each of them harbored two isolates; additionally, four isolates did not cluster at this similarity level. Isolate W4 was carefully chosen as the most promising pigment and antimicrobial agent's producer; the produced pigment was extracted and optimized by statistical experiments (PBD & BBD) and was tested for its anti-inflammatory activity. The results showed anti-inflammatory effect and prevented the denaturation of BSA protein at a concentration much higher than the safe dose and increased with increasing the pigment concentration. CONCLUSION: Marine actinomycetes play a vital role in the production of novel and important economic metabolites that have many industrial and pharmaceuticals applications. Streptomyces genera are the most important actinomycetes that produce important metabolites as previously reported.