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Biochemistry, Genetics and Molecular Biology » "Carotenoids", book edited by Dragan J. Cvetkovic and Goran S. Nikolic, ISBN 978-953-51-3212-7, Print ISBN 978-953-51-3211-0, Published: June 14, 2017 under CC BY 3.0 license. © The Author(s).

Chapter 8

Biotechnological Production of Carotenoids and Their Applications in Food and Pharmaceutical Products

By Ligia A. C. Cardoso, Susan G. Karp, Francielo Vendruscolo, Karen Y. F. Kanno, Liliana I. C. Zoz and Júlio C. Carvalho
DOI: 10.5772/67725

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Biotechnological Production of Carotenoids and Their Applications in Food and Pharmaceutical Products

Ligia A. C. Cardoso1, Susan G. Karp2, Francielo Vendruscolo3, Karen Y. F. Kanno1, Liliana I. C. Zoz2 and Júlio C. Carvalho2
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Pigments can be divided into four categories: natural, nature-identical, synthetic, and inorganic colors. Artificial colorants are the most used in food and pharmaceutical industries because of their advantages related to color range, price, resistance to oxygen degradation, and solubility. However, many natural pigments present health-promoting activities that make them an interesting option for human use and consumption. Natural colorants are derived from sources such as plants, insects, and microorganisms. Carotenoids are natural pigments with important biological activities, such as antioxidant and pro-vitamin A activity, that can be either extracted from plants and algae or synthesized by various microorganisms, including bacteria, yeasts, filamentous fungi, and microalgae. Advantages of microbial production include the ability of microorganisms to use a wide variety of low cost substrates, the better control of cultivation, and the minimized production time. After fermentation, carotenoids are usually recovered by cell disruption, solvent extraction, and concentration. Subsequent purification steps are followed depending on the application. The most prominent industrial applications of carotenoids, considering their health benefits, are in the food, feed, and pharmaceutical industries.

Keywords: biotechnology, natural pigments, microbial carotenoids, downstream, industrial applications

1. Introduction

Color has a great influence on the appearance, processing, and acceptance of food products, textiles and pharmaceutical products. The first quality impact by which consumers make the decision to purchase a product is its visual appearance.

Food colorants can be divided into four categories: natural, nature-identical, synthetic, and inorganic colors [1]. The production of synthetic coloring agents and other chemicals used as food additives is under increasing pressure due to a renewed interest in the use of natural products and the strong interest in minimizing the use of chemical processes [2]. Since the number of permitted synthetic colorants has decreased because of undesirable toxic effects including mutagenicity and potential carcinogenicity, interest focuses on the development of food grade pigments from natural sources [35].

Natural pigments are derived from sources such as plants, insects, and microorganisms. Algae and microalgae, bacteria, fungi, and yeasts are organisms commonly found in nature that can produce natural pigments in different color spectra, such as violacein, phycocyanin, monascins, flavins, quinones, and carotenoids.

Carotenoids represent one of the most important groups of natural pigments, they are responsible for the yellow, orange, red, and purple colors in a wide variety of plants, animals, and microorganisms [6]. They are lipid-soluble, commercially, and biotechnologically significant pigments produced from various organisms such as plants [7], algae and microalgae [812], bacteria [1315], fungi [1620], and yeasts [4, 2125].

Pigments from natural sources have been obtained since long time ago, and their attractiveness has increased due to the toxicity problems caused by the synthetic pigments [2628]. Carotenoids are obtained industrially by chemical synthesis or extraction from plants or algae; however, there has been an increasing interest in biotechnological processes for carotenoids production [29]. The pigments from microbial sources are a good alternative to obtain natural colorants for industrial uses.

The biotechnological production of carotenoids has advantages related to the diversity of microorganisms in nature, versatility in the use of substrates and agro-industrial wastes and the possibility to control operating conditions such as pH, temperature, dissolved oxygen, and light intensity; also, biomass from other bioprocesses can be submitted to the extraction of carotenoids. The production of microbial carotenoids has become a potential alternative for the replacement of artificial pigments, even with technological, economic, and legislation limitations.

Studies have demonstrated that carotenoids play an essential role for the maintenance of living bodies. In plants, carotenoids play an important role in photosynthesis, acting as light-harvesting pigments and protectors against photo-oxidation. In foods, carotenoids confer yellow, orange, or red color, serve as precursors of aroma compounds, and, as natural antioxidants, may help to extend the shelf-life [30, 31]. In humans, carotenoids have been associated with the reduction of the risk of developing chronic diseases such as cancer, cardiovascular diseases, high levels of cholesterol, cataract, and macular degeneration, aside from the pro-vitamin A activity of some of these compounds [3133]. This is important because in the developed world, as life expectancy increases and the birth rate declines, the demand for solutions focusing on longevity and life quality increases too. The number of people aged >60 years is expected to account for approximately one-fifth of the world’s population by 2050 [34].

2. Biotechnological production of carotenoids

2.1. Carotenoids diversity

Carotenoids are lipid-soluble pigments, colored from yellow to red, with a basic structure consisting in a tetraterpene with a series of conjugated double bonds. They can have only carbon and hydrogen in their structures or have one or more oxygen atoms, being classified as xanthophylls. The majority of carotenoids are C40 terpenoids, which act as membrane-protective antioxidants scavenging O2 and peroxyl radicals [35].

There are more than 700 types of carotenoids described and only about 50 are precursors of vitamin A. Carotenoids can reduce risks for degenerative diseases such as cancer, cardiovascular diseases, macular degeneration, and cataract. The biological activities, specially the antioxidant properties, depend on their chemical structure: number of conjugated double bonds, structural end-groups, and oxygen-containing substituents [36].

Carotenoids occur in photosynthetic systems of higher plants, algae, and phototrophic bacteria. In plants, carotenoids are embedded in the membranes of chloroplasts and chromoplasts. The colors of these pigments are masked by chlorophyll, but they contribute to the bright colors of many flowers and fruits [37].

Nonphotosynthetic organisms, as some bacteria and fungi, present carotenoids as protectors against photo-oxidative damage, a way of protection in growth conditions with light and abundant air. The main carotenoids produced by fungi are β-carotene, torulene, torularhodin, and astaxanthin [38]. Bacteria have been reported as producers of cantaxanthin mainly. The microalgae are producers of lutein, β-carotene, and astaxanthin [35].

Animals usually present carotenoids provenient from their diet. Marine animals that feed on algae or on products rich in carotenoids may exhibit the coloration of these pigments, as the salmon fish. The color of the feathers of some birds also comes from a diet rich in carotenoids, as flamingos [39].

The industrial production of carotenoids by plants is dependent on the season and geographic variability, and these cannot always be controlled. The chemical synthesis of carotenoids generates wastes that can cause damage to the environment and resistance by the consumers. Because of this, the biotechnological resources are becoming more interesting. The microbial production of carotenoids can be performed using low-cost substrates or substrates that are residues from industrial processes, like molasses, resulting in lower costs of production [40]. All conditions of this kind of production can be controlled and optimized, especially knowing the metabolic route of each microorganism utilized.

Carotenoids are intracellular products, and a process to increase their accessibility at the downstream stage is necessary. The techniques most used combine physical and chemical methods like maceration and contact with organic solvents [4].

2.2. Main carotenoid biosynthesis pathways

Carotenoids are usually produced from the building blocks geranyl geranyl diphosphate (GGPP) and farnesyl diphosphate (FPP), like other secondary metabolites such as sesquiterpenoids and steroids. The most common pathway is the condensation of 2 GGPP units into prephytoene diphosphate and then to phytoene, a 40-carbon polyunsaturated precursor which is colorless. This precursor is converted into lycopene and then into several derived carotenoids such as β-carotene and oxidized derivatives such as lutein. The condensation of two units of FPP leads to 30-carbon precursors that are converted to steroids or apocarotenoids such as staphyloxanthin [41, 42]. Apocarotenoids can also be produced by oxidative cleavage of carotenoids. Figure 1 presents a simplified carotenoid biosynthesis pathway.


Figure 1.

Biosynthesis pathways of common carotenoids. Source: Adapted from Ref. [38] with permission.

Most carotenoids present maximal absorption in the violet to green region of the visible spectrum, so these substances appear as red to yellow pigments. Table 1 shows the carotenoids with permitted food use according to the Food and Drug Administration (FDA) and the Food and Agriculture Organization (FAO).

Additive/sourceColorMain componentInternational Numbering System
Algae meal, driedGreen to redMixture of carotenoids, xanthophyll, and chlorophylls
Astaxanthin and astaxanthin dimethyldisuccinate (several microorganisms)Orange-redAstaxanthin
β-apo-8′-carotenal (from carrot oil)Reddish orangeAll-trans-β-apo-8′-carotenal160e, 160f
β-carotene, synthetic and natural: from vegetables, Blakeslea trispora and Dunaliella salinaOrangeβ-carotene160a(i), 160a(ii)
Canthaxanthin; most of the pigment used in feeds is syntheticOrange pinkβ-carotene-4,4′-dione (canthaxanthin)161g
Carrot oilOrange to yellow160a(ii)
Gardenia red and yellowRed, yellowCrocin, crocetin
Haematococcus algae mealOrange-redAstaxanthin
Lycopene, tomato extract (i) or concentrate (ii) or from Blakeslea trispora (iii)RedLycopene160d
Lutein (bras), from marigold oleoresinLutein161b
Marigold colorYellowLutein
Orchil dyesRedOrcein
Paprika and paprika oleoresinRedCapsanthin, capsorubin160c
Paracoccus pigmentRedAstaxanthin
Phaffia yeastOrange-redAstaxanthin esters
SaffronYellow to orangeα-crocin164
Tagetes (Aztec marigold) meal and extractYellow to orangeLutein161b

Table 1.

Carotenoids and carotenoid-rich products used as food color additives.

Sources: Compiled from the FDA Color Additive Status List [] and from the Combined Compendium of Food Additive Specifications [].

2.3. Carotenoid sources

The most common sources for natural carotenoids for food and cosmetic use are plants, although microorganism biomass is becoming more common as a source for these substances. Table 2 illustrates some commercial sources for microorganism-based carotenoids.

MicroorganismMoleculeCulture medium*Xmax (g/L)Pmax (mg/L)Conc. (mg/g)**μx (h−1)References
Blakeslea trispora (fungus)β-caroteneCorn steep liquor20800400.022[43]
Blakeslea trisporaβ-caroteneWhey813601700.023[44]
Sporobolomyces roseus (yeast)β-caroteneReconstituted whey4.712.580.55[40]
Rhodotorula glutinis (yeast)β-carotenePotato extract5.701.080.19[40]
Dietzia natronolimnaea (bacterium)CanthaxanthinWhey3.292.870.870.020[45]
Phaffia rhodozyma (yeast)AstaxanthinCassava residues8.62.980.350.060[46]
Sporobolomyces ruberrimus (yeast)TorularhodineTechnical glycerol303.70.120.040[47]
Chlorella zofingiensis (microalga)AstaxanthinBBM with glucose10.210.031[48]
Coelastrella striolata (microalga)Canthaxanthin
Coccomyxa onubensis (microalga)β-carotene
Haematococcus pluvialis (microalga)AstaxanthinBBM2.213.5[51]
Chlorella zofingiensisAstaxanthinBristol, modified101.250.043[52]
Dunaliella salina (microalga)β-carotenef214***0.55[53]
Haematococcus pluvialisAstaxanthinStandard312–150.56[54]
Muriellopsis sp. (microalga)LuteinArnon, modified5.376.510.17–0.23[55]
Haematococcus pluvialis (wild-type)
Haematococcus pluvialis (mutant)
AstaxanthinNIES medium1.6

Paracoccus carotinifaciens (bacterium)Astaxanthin CanthaxanthinGlucose and peptone based25–40[57]

Table 2.

Main sources for concentrated carotenoids.

* Except where specified, these are mineral-based media. Recipes may be found at UTEX, SAG, or CCMP collections web sites.

** Milligrams of carotenoids per gram of biomass.

*** Estimated. The original reference reports 28.1 mg/L carotenoids.

Xmax—maximum biomass concentration; Pmax—maximum carotenoids concentration; μX—biomass production rate.

Source: Adapted from Ref. [58].

2.4. General downstream operations for carotenoid production

Carotenoids are nonpolar molecules that accumulate intracellularly in plant tissues and microorganisms. Therefore, the production usually consists in a biomass pretreatment that may accelerate the dissolution of these substances, followed by a solid-liquid extraction (leaching) with a suitable, low-polarity solvent. The resulting solution can be a final product, can be desolventized, and can be further purified, depending on the use intended for the extract. Figure 2 illustrates the main steps in the production of carotenoids.


Figure 2.

Main steps in carotenoids production.

The first step in carotenoid production is the pretreatment of the raw biomass, usually by drying and milling. Drying is convenient because it reduces the weight of the material to be processed, facilitates the access for solvents to the biomass, and reduces contaminants that could be extracted in water micelles with the solvent. The milling step is also important because it increases the surface area of the biomass matrix, facilitating contact with the solvent. In the case of tough-walled organisms, chemical or mechanical cell disruption may be done prior to drying. Fine milling of the dry biomass is less common.

The dry biomass is then extracted using a nonpolar solvent such as hexane or a vegetable oil, for the dissolution of carotenoids. A higher polarity solvent such as acetone can be used for the extraction of xanthophylls. In both cases, lipids are extracted in the mix. This extraction is an equilibrium operation; therefore, the final concentration in the solvent affects the extraction efficiency. Following extraction, the solution containing carotenoids must be concentrated and desolventized. This is why low boiling point solvents, which are easy to evaporate, are more common extractants than oils.

The carotenoids in the concentrated extract may be purified or not, depending on the intended use. For example, β-carotene that will be used as a vitamin A precursor must be purified, while paprika oleoresin is a mixture of carotenoids used mainly as a color and flavor additive and needs no further purification. In general, for carotenoids used as color additives, it is enough to concentrate the extract because (1) the tinctorial strength of the molecules is large—therefore, the additive is added at a low concentration to the formulated product and (2) the sources used are generally regarded as safe (GRAS), and the molecules extracted with the carotenoid are harmless in the concentrations used.

In the case of purified carotenoids, the operations to be used—adsorption, chromatography, crystallization, etc.—depend largely on the properties of the target molecule and the contaminants in the mixture, such as melting point, polarity, solubility, etc. All sorts of nonpolar compounds are extracted with the solvent, such as neutral and slightly polar lipids, steroids, and waxes. The differences in the properties of the carotenoid and the contaminants will be explored in the purification strategy.

Following extraction and purification, the carotenoid must be formulated for further application. This formulation will also depend on the intended use. The formulation may be as simple as adding an antioxidant such as butylated hydroxytoluene (BHT) or butylated hydroxyanisole (BHA) to the extract or may be more complex, such as emulsifying the carotenoid as an oil-in-water product for use in polar matrixes such as juices.

3. Industrial application of carotenoids as additives in food, feed and pharmaceutical products

Because of the rising of health concerns by consumers, the demand for carotenoids as natural coloring products is growing. Beta-carotene, astaxanthin, canthaxanthin, lycopen, and lutein are the most required and valuable carotenoids, and they are currently used by the food, feed, and cosmetic industries (Table 3). The use of carotenoids is regulated by the legislation of each country that specifies the source, purity, product, and quantities of the colorant that can be used [59].

LuteinYellowPoultry feed; functional nutrientAntioxidant[60]
CanthaxanthinOrangePoultry feed; fish feed; cosmeticAntioxidant, anticancer[61]
LycopeneRedSupplement in functional foods; additive in cosmeticsAntioxidant, anticancer[62]
β-caroteneOrange-redNutraceutical; cosmetic; animal feed industriesAntioxidant, anticancer, precursor of vitamin A[63]
AstaxanthinPink-redFish feed; cosmetic industryAntioxidant, photoprotectant, anticancer, anti-inflammatory[64]

Table 3.

Carotenoids' colors, applications and biological activities.

According to BBC Research [65], the carotenoid global market in 2014 was of US$ 1.5 billion, this value is increasing year by year and is expected to reach US$ 1.8 billion in 2019, with an annual growth rate of 3.9%. Beta-carotene, the carotenoid of highest value, had a global market of US$ 233 million in 2010, which is expected to reach US$ 309 million by 2018. Astaxanthin, due to its powerful antioxidant activity, is the third carotenoid in terms of high added value, with a global market size of US$ 225 million, estimated to increase to US$ 253 million by 2018.

3.1. Importance and use of carotenoids in food products

Commercial food products using carotenoids are expanding, and the greatest demand is in the Asian continent. The pigment is extracted from microalgae such as Chlorella, Dunaliella, Haematococcus [66, 67], from the cyanobacterium Spirulina [68], and from the fungus Monascus [69].

In Asia, the production red koji dates of hundreds of years and uses the fermentation of rice by Monascus to produce the typical reddish color. These red pigments are also used as food colorants for wine, red soy cheese, meat, and by-products of meat and fish [26]. The French cheese named vieux-pan contains the carotenoid produced by Brevibacterium linens due to its orange-red-brown color that improves the sensory quality of the product [70]. In Russia, infant formulas are enriched with natural pigments such as lutein, which is present in breast milk, in order to improve children's health [71].

Nutraceutical food products have also been applied in bakery products and pasta. In Japan, Undaria pinnatifida (wakame), an edible seaweed rich in fucoxanthin, is commercialized as an ingredient for pasta [72]. In India, a pasta containing fucoxanthin as an ingredient to improve its biofunctional and nutritional qualities was developed [73].

3.2. Importance and applications of carotenoids in the pharmaceutical industry

Besides the use of nutraceutical foods as a form of prevention and treatment of diseases, the administration of the bioactive compounds in their concentrated form is also a possibility for promoting health. The transport of carotenoids occurs from the intestinal mucosa to the blood vessels carried by lipoproteins [74]. Carotenoids functional properties are related to reactions such as oxidation, reduction, hydrogen abstraction, and addition in biological membranes, and their antioxidant power is fundamental for cell protection against free radicals and singlet oxygen formed in tissues [75].

Some carotenoids are precursors of vitamins, and they also present activities such as anti-inflammatory, antioxidant, immunomodulatory, anticancer, for cardiovascular therapy and neurodegenerative diseases [76], and anti-obesity [77]. The carotenoids included as pro-vitamin A are β-carotene, α-carotene, and cryptoxanthin. Vitamin A is an essential nutrient for operation and maintenance of biological functions including vision, reproduction, and immunity [78]. Beta-carotene is present in blood and tissues, which is associated with antioxidant activity and concomitantly with other carotenoids or antioxidants can enhance their activity against free radicals. However, it can bring health risk at high doses [79].

Carotenoids, acting as antioxidants eliminating free radicals, can modulate the risk of developing chronic diseases by inhibiting reactions mediated by reactive oxygen species (ROS). Reactive species are produced during cellular metabolism as a defense to infectious and chemical agents that may cause damage to DNA, proteins, and tissues, contributing to the development of chronic diseases such as diabetes, Parkinson’s, Alzheimer’s, cardiovascular diseases, and cancer [80].

In addition to the antioxidant properties, carotenoids exhibit anti-inflammatory activities owing to the protective effects of phytochemicals such as lutein and astaxanthin. Astaxanthin has been shown to inhibit the production of pro-inflammatory mediators such as nitric oxide (NO) in macrophages, to increase the level of inflammatory cytokines, and to reduce oxidative stress. Neuroprotective effect, reduced neuroinflammation, improvement of insulin signals, and reduction of lipid levels were also verified [81].

Inhibition of cell proliferation of colon cancer cells by the use of Neochloris oleoabundans carotenoids was observed, enabling its use as a functional food additive or nutraceutical with potential for the prevention of colon cancer [82]. Beta-carotene, astaxanthin, and capsanthin demonstrated antiproliferative effects on leukemic K562 cells [83]. Studies indicated that the simultaneous use of different carotenoids was efficient against liver cancer. Patients were administered with β-cryptoxanthin-enriched mandarin orange juice and capsules of a carotenoids mixture-containing lutein, β-cryptoxanthin, lycopene, zeaxanthin, and fucoxanthin. Analyses of DNA array and protein-antibody array showed that the carotenoids interferred in the induction of genes such as p16 and p73 [84].

4. Conclusion and final remarks

There are many advantages related to the use of carotenoids instead of artificial pigments in food products and for pharmaceutical applications. Their biological properties such as antioxidant, anti-inflammatory, antitumoral, and pro-vitamin A activities contribute to the quality of the product and to the consumer’s health. Among the production strategies, microbial synthesis is considered advantageous, and the downstream techniques usually involve cell disruption, solvent extraction, concentration, and purification, when necessary. Several researches have proved the beneficial effects of carotenoids on health, so they can meet the demand for solutions focusing on longevity and life quality.


1 - Aberoumand A. A review article on edible pigments properties and sources as natural biocolorants in foodstuff and food industry. World Journal of Dairy & Food Sciences. 2011;6:71-78.
2 - Domínguez-Espinosa RM, Webb C. Submerged fermentation in wheat substrates for production of Monascus pigments. World Journal of Microbiology and Biotechnology. 2003;19:329-336. DOI: 10.1023/A:1023609427750
3 - Sabater-Vilar M, Maas RFM, Fink-Gremmels J. Mutagenicity of commercial Monascus fermentation products and the role of citrinin contamination. Mutation Research. 1999;444:7-16. DOI: 10.1016/S1383-5718(99)00095-9
4 - Pennacchi MGC, Rodrígues-Fernández DE, Vendruscolo F, Maranho LT, Marc I, Cardoso LAC. A comparison of cell disruption procedures for the recovery of intracellular carotenoids from Sporobolomyces ruberrimus H110. International Journal of Applied Biology and Pharmaceutical Technology. 2015;6:136-143.
5 - Vendruscolo F, Bühler RMM, Carvalho JC, Oliveira D, Moritz DE, Schmidell W, Ninow JL. Monascus: A reality on the production and application of microbial pigments. Applied Biochemistry and Biotechnology. 2016;178:211-223. DOI: 10.10007/s12010-015-1880-z
6 - Oliver J, Palou A. Chromatographic determination of carotenoids in foods. Journal of Chromatography A. 2000;881:543-555.
7 - Hanson P, Yang RY, Chang LC, Ledesma L, Ledesma D. Mint: Carotenoids, ascorbic acid, minerals, and total glucosinolates in choysum (Brassica rapa cvg. parachinensis) and kailaan (B. oleraceae Alboglabra group) as affected by variety and wet and dry season production. Journal of Food Composition and Analysis. 2011;24:950-962. DOI: 10.1016/j.jfca.2011.02.001
8 - Rodrigues DB, Flores EMM, Barin JS, Mercadante AZ, Jacob-Lopes E, Zepka LQ. Production of carotenoids from microalgae cultivated using agroindustrial wastes. Food Research International. 2014;65:144-148. DOI: 10.1016/j.foodres.2014.06.037
9 - Přibyl P, Cepák V, Kaštánek P, Zachleder V. Elevated production of carotenoids by a new isolate of Scenedesmus sp. Algal Research. 2015;11:22-27. DOI: 10.1016/j.algal.2015.05.020
10 - Chen L, Zhang L, Liu T. Concurrent production of carotenoids and lipid by a filamentous microalga Trentepohlia arborum. Bioresource Technology. 2016;214:567-573. DOI: 10.1016/j.biortech.2016.05.017
11 - Liu J, Mao X, Zhou W, Guarnieri MT. Simultaneous production of triacylglycerol and high-value carotenoids by the astaxanthin-producing oleaginous green microalga Chlorella zofingiensis. Bioresource Technology. 2016;214:319-327. DOI: 10.1016/j.biortech.2016.04.112
12 - Tsai HP, Chuang LT, Chen CNN. Production of long chain omega-3 fatty acids and carotenoids in tropical areas by a new heat-tolerant microalga Tetraselmis sp. DS3. Food Chemistry. 2016;192:682-690. DOI: 10.1016/j.foodchem.2015.07.071
13 - Fang CJ, Ku KL, Lee MH, Su NW. Mint: Influence of nutritive factors on C50 carotenoids production by Haloferax mediterranei ATCC 33500 with two-stage cultivation. Bioresource Technology. 2010;101:6487-6493. DOI: 10.1016/j.biortech.2010.03.044
14 - Peter-Wendisch P, Götker S, Heider SAE, Reddy K, Nguyen AQ, Stansen KC, Wendisch VF. Engineering biotin prototrophic Corynebacterium glutamicum strains for amino acid, diamine and carotenoid production. Journal of Biotechnology. 2014;192:346-354. DOI: 10.1016/j.jbiotec.2014.01.023
15 - Autenrieth C, Ghosh R. Random mutagenesis and overexpression of rhodopin-3,4-desaturase allows the production of highly conjugated carotenoids in Rodospirillum rubrum. Archives of Biochemistry and Biophysics. 2015;572:134-141. DOI: 10.1016/
16 - Goodwin TW. Fungal carotenoids. Botanical Review. 1952;18:291-316.
17 - El-Jack M, Mackenzie A, Bramley PM. The photoregulation of carotenoid biosynthesis in Aspegillus giganteus mut. alba. Planta. 1998;174:59-66.
18 - Denter J, Rehm HJ, Bisping B. Changes in the contents of fat-soluble vitamins and provitamins during tempo fermentation. International Journal of Food Microbiology. 1998;45:129-134.
19 - Iturriaga EA, Papp T, Breum J, Arnau J, Eslava AP. Strain and culture conditions improvement for b-carotene production in Mucor. In: Microbial Processes and Products, Methods in Biotechnology series. 1st ed. Humana Press; 2005. pp. 239-256. DOI: 10.1385/1-59259-847-1:239.
20 - Csernetics A, Nagy G, Iturriaga EA, Szekeres A, Eslava AP, Vágvölgyi C, Papp T. Expression of three isoprenoid biosynthesis genes and their effects on the carotenoid production of the zygomycete Mucor circinelloides. Fungal Genetics and Biology. 2011;48:696-703. DOI: 10.1016/j.fgb.2011.03.006
21 - Valduga E, Ribeiro AHR, Cence K, Coilet R, Tiggemann L, Zeni J, Toniazzo G. Carotenoids production from a newly isolated Sporidiobolus pararoseus strain using agroindustrial substrates. Biocatalysis and Agricultural Biotechnological. 2014;3:207-213. DOI: 10.1016/j.bcab.2013.10.001
22 - Dias C, Sousa S, Caldeira J, Reis A, Silva TL. New dual-stage pH control fed-batch cultivation strategy for the improvement of lipids and carotenoids production by the red yeast Rhodosporidium toruloides NCYC 921. Bioresource Technology. 2015;189:309-318. DOI: 10.1016/j.biortech.2015.04.009
23 - Cardoso LAC, Jäckel S, Karp SG, Framboisier X, Chevalot I, Marc I. Improvement of Sporobolomyces ruberrimus carotenoids production by the use of raw glycerol. Bioresource Technology. 2016;200:374-379. DOI: 10.1016/j.biortech.2015.09.108
24 - Odoñez MC, Raftery JP, Jaladi T, Chen X, Kao K, Karim MN. Modelling of batch kinetics of aerobic carotenoid production using Saccharomyces cerevisiae. Biochemical Engineering Journal. 2016;114:226-236. DOI: 10.1016/j.bej.2016.07.004
25 - Yoo AY, Alnaeeli M, Park JK. Production control and characterization of antibacterial carotenoids from the yeast Rhodotorula mucilaginosa AY-01AH. Process Biochemistry. 2016;51:463-473. DOI: 10.1016/j.procbio.2016.01.008
26 - Dufossé L, Galaup P, Yaron A, Arad SM, Blanc P, Murthy KNC, Ravishankar G. Microorganisms and microalgae as sources of pigments for food use: A scientific oddity or an industrial reality? Trends in Food Science and Technology. 2005;16:389-406. DOI: 10.1016/j.tifs.2005.02.006
27 - Dufossé, L. Production of food grade pigments. Food Technology and Biotechnology. 2006;44:313-321.
28 - Kumar A, Vishwakarma HS, Singh J, Dwivedi S, Kumar M. Microbial pigments: Production and their applications in various industries. International Journal of Pharmaceutical, Chemical and Biological Sciences. 2015;5:203-212.
29 - Valduga E, Tatsch PO, Tiggemann HT, Toniazzo G, Zeni J, Luccio M. Produção de carotenoides: Microrganismos como fonte de pigmentos naturais. Química Nova. 2009;32:2429-2436. DOI: 10.1590/S0100-40422009000900036
30 - Ruiz-Sola MA, Rodríguez-Concepción M. Carotenoid biosynthesis in Arabidopsis: A colorful pathway. In: The Arabidopsis Book: American Society of Plant Biologists. 1st ed. 2012. 29. DOI: 10.1199/tab.0158
31 - Rodriguez-Amaya DB. Status of carotenoid analytical methods and in vitro assays for the assessment of food quality and health effects. Current Opinion in Food Science. 2015;1:56-63. DOI: 10.1016/j.cofs.2014.11.005
32 - Krinsky NI, Johnson E. Carotenoid actions and their relation to health and disease. Molecular Aspects of Medicine. 2005;26:459-516. DOI: 10.1016/j.mam.2005.10.001
33 - Woodside JV, McGrath AJ, Lyner N, McKinley MC. Carotenoids and health in older people. Maturitas. 2015;80:63-68. DOI: 10.1016/j.maturitas.2014.10.012
34 - WHO – World Health Organization. Ageing and Life Course [Internet]. 2012. Available from: [Accessed: 2016-08-08].
35 - Mata-Gómez LC, Montañez JC, Méndez-Zavala A, Aguilar CN. Biotechnological production of carotenoids by yeasts: An overview. Microbial Cell Factories. 2014;13:12. DOI: 10.1186/1475-2859-13-12
36 - Rodrigues E, Mariutti LRB, Chisté RC, Mercadante AZ. Development of a novel micro-assay for evaluation of peroxyl radicalscavenger capacity: Application to carotenoids and structure-activity relationship. Food Chemistry. 2006;135:2103-2111. DOI: 10.1016/j.foodchem.2012.06.074
37 - Bartley GE, Scolnik PA. Plant carotenoids: Pigments for photoprotection, visual attraction and human health. The Plant Cell. 1995;7:1027-1038. DOI: 10.1105/tpc.7.7.1027
38 - Zoz L, Carvalho JC, Soccol VT, Casagrande CC, Cardoso L. Torularhodin and torulene: Bioproduction, properties and prospective applications in food and cosmetics – A review. Brazilian Archives of Biology and Technology. 2015;58:278-288. DOI: 10.1590/S1516-8913201400152
39 - Hill GE, Inouye CY, Montgomerie R. Dietary carotenoids predict plumage coloration in wild house finches. Proceeding of the Royal Society B: Biological Sciences. 2002;269:1119-1124. DOI: 10.1098/rspb.2002.1980
40 - Marova I, Carnecka M, Halienova A, Certik M, Dvorakova T, Haronikova A. Use of several waste substrates for carotenoid-rich yeast biomass production. Journal of Environmental Management. 2012;95:338-342. DOI: 10.1016/j.jenvman.2011.06.018
41 - Lin FY, Liu CI, Liu YL, Zhang Y, Wang K, Jeng WY, Ko TP, Cao R, Wang AH, Oldfield E. Mechanism of action and inhibition of dehydrosqualene synthase. Proceedings of the National Academy of Sciences. 2010;107:21337-21342. DOI: 10.1073/pnas.1010907107
42 - Farré G, Sanahuja G, Naqvi S, Bai C, Capell T, Zhu C, Christou P. Travel advice on the road to carotenoids in plants. Plant Science. 2010;179:28-48. DOI: 10.1016/j.plantsci.2010.03.009
43 - Papaioannou EH, Liakopoulou-Kyriakides M. Substrate contribution on carotenoids production in Blakeslea trispora cultivations. Food and Bioproducts Processing. 2010;8:305-311. DOI: 10.1016/j.fbp.2009.03.001
44 - Varzakakou M, Roukas T, Kotzekidou P. Mint: Effect of the ratio of (+) and (−) mating type of Blakeslea trispora on carotene production from cheese whey in submerged fermentation. World Journal of Microbiology Biotechnology. 2010;26:2151-2156. DOI: 10.1007/s11274-010-0398-3
45 - Khodaiyan F, Razavi SH, Mousavi SM. Optimization of canthaxanthin production by Dietzia natronolimnaea HS-1 from cheese whey using statistical experimental methods. Biochemical Engineering Journal. 2008;40:415-422. DOI: 10.1016/j.bej.2008.01.016
46 - Yang J, Tan H, Yang R, Sun X, Zhai H, Li K. Astaxanthin production by Phaffia rhodozyma fermentation of cassava residues substrate. Agricultural Engineering International. 2011;13:1-6.
47 - Razavi SH, Marc I. Effect of temperature and pH on the growth kinetics and carotenoid production by Sporobolomyces ruberrimus H110 using technical glycerol as carbon source. Iranian Journal of Chemistry and Chemical Engineering. 2006;25:59-64.
48 - Ip PF, Chen F. Production of astaxanthin by the green microalga Chlorella zofingiensis in the dark. Process Biochemistry. 2005;40:733-738. DOI: 10.1016/j.procbio.2004.01.039
49 - Abe K, Hattori H, Hirano M. Accumulation and antioxidant activity of secondary carotenoids in the aerial microalga Coelastrella striolata var. multistriata. Food Chemistry. 2007;100:656-661. DOI: 10.1016/j.foodchem.2005.10.026
50 - Vaquero I, Ruiz-Domínguez C, Márquez M, Vílchez C. Cu-mediated biomass productivity enhancement and lutein enrichment of the novel microalga Coccomyxa onubensis. Process Biochemistry. 2012;47:694-700. DOI: 10.1016/j.procbio.2012.01.016
51 - Harker M, Tsavalos A, Young AJ. Factors responsible for astaxanthin formation in the chlorophyte Haematococcus pluvialis. Bioresource Technology. 1996;55:207-217. DOI: 10.1016/0960-8524(95)00002-X
52 - Ip PF, Wong KH, Chen F. Enhanced production of astaxanthin by the green microalga Chlorella zofingiensis in mixotrophic culture. Process Biochemistry. 2004;39:1761-1766. DOI: 10.1016/j.procbio.2003.08.003
53 - Kleinegris DMM, Janssen M, Brandenburg WA, Wijffels RH. Continuous production of carotenoids from Dunaliella salina. Enzyme and Microbial Technology. 2011;48:253-259. DOI: 10.1016/j.enzmictec.2010.11.005
54 - Garcıa-Malea MC, Brindley C, Del Río E, Acien FG, Fernandez JM, Molina E. Modeling of growth and accumulation of carotenoids in Haematococcus pluvialis as a function of irradiance and nutrients supply. Biochemical Engineering Journal. 2005;26:107-114. DOI: 10.1016/j.bej.2005.04.007
55 - Del Campo JA, Moreno J, Rodrıguez H, Vargas MA, Rivas J, Guerrero MJ. Carotenoid content of chlorophycean microalgae: Factors determining lutein accumulation in Muriellopsis sp. (Chlorophyta). Journal of Biotechnology. 2000;76:51-59. DOI: 10.1016/S0168-1656(99)00178-9
56 - Hong ME, Choi SP, Park YI, Kim YK, Chang WS, Kim BW, Sim SJ. Astaxanthin production by a highly photosensitive Haematococcus mutant. Process Biochemistry. 2012;47:1972-1979. DOI: 10.1016/j.procbio.2012.07.007
57 - Hirschberg J, Harker M. Carotenoid-Producing Bacterial Species and Process for Production of Carotenoids Using Same. United States Patent 5,935,808. August 10, 1999.
58 - De Carvalho JC, Cardoso LC, Ghiggi V, Woiciechowski AL, de Souza Vandenberghe LP, Soccol CR. Microbial pigments. In: Brar SK, Dhillon GS, Soccol CR, editors. Biotransformation of Waste Biomass into High Value Biochemicals. Springer: New York; 2014. pp. 73-97. ISBN: 978-1-4614-8005-1
59 - Jaswir I, Noviendri D, Hasrini RF, Octavianti F. Carotenoids: Sources, medicinal properties and their application in food and nutraceutical industry. Journal of Medicinal Plants Research. 2011;5:7119-7131. DOI: 10.5897/JMPRx11.011
60 - Lin JH, Lee DJ, Chang JS. Lutein production from biomass: Marigold flowers versus microalgae. Bioresource Technology. 2015;184:421-428. DOI: 10.1016/j.biortech.2014.09.099
61 - Ravaghi M, Razavi SH, Mousavi SM, Sinico C, Fadda AM. Stabilization of natural canthaxanthin produced by Dietzia natronolimnaea HS-1 by encapsulation in niosomes. Food Science and Technology. 2016;73:498-504. DOI: 10.1016/j.lwt.2016.06.027
62 - Hernandez-Almanza A, Montañez J, Martínez G, Aguilar-Jimenez A, Contreras-Esquivel JC, Aguilar C N. Lycopene: Progress in microbial production. Trends in Food Science & Technology. 2016;56:142-148. DOI: 10.1016/j.tifs.2016.08.013
63 - Jing K, He S, Chen T, Lu Y, Ng I-S. Enhancing beta-carotene biosynthesis and gene transcriptional regulation in Blakeslea trispora with sodium acetate. Engineering Journal. 2016;114:10-17. DOI: 10.1016/j.bej.2016.06.015
64 - Panis G, Carreon JR. Commercial astaxanthin production derived by green alga Haematococcus pluvialis: A microalgae process model and a techno-economic assessment all through production line. Algal Research. 2016;18:175-190. DOI: 10.1016/j.algal.2016.06.007
65 - BBC Research. The Global Market for Carotenoids [Internet]. 2015. Available from: [Accessed: 2017-01-18]
66 - Spolaore P, Joaniss-Cassan C, Duran E, Isambert A. Commercial applications of microalgae. Journal of Bioscience and Bioengineering. 2006;101:87-96. DOI: 10.1263/jbb.101.87
67 - Raja R, Haemaiswarya S, Rengasamy R. Exploitation of Dunaliella for β-carotene production. Applied Microbiology and Biotechnology. 2007;74:517-523. DOI: 10.1007/s00253-006-0777-8d
68 - Singh RP. Spirulina: Health food for complete nutrition. Biotech Today. 2013;3:48-51. DOI: 10.5958/j.2322-0996.3.1.009
69 - Lian X, Liu L, Dong S, Wu H, Zhao J, Han Y. Two new monascus red pigments produced by Shandong Zhonghui Food Company in China. European Food Research and Technology. 2014;240:719-724. DOI: 10.1007/s00217-014-2376-8
70 - Galaup P, Sutthiwong N, Leclercq-Perlat MN, Valla A, Caro Y, Fouillaud M, Guérard F, Dufossé L. First isolation of Brevibacterium sp. pigments in the rind of an industrial red-smear-ripened soft cheese. Society of Dairy Technology. 2015;68:144-147. DOI: 10.1111/1471-0307.12211
71 - Kon IY, Gmoshinskaya MV, Safronova AI, Alarcon P, Vandenplas Y. Growth and tolerance assessment of a lutein-fortified infant formula. Journal Pediatric Gastroenterology Hepatology Nutrition. 2014;17:104-111. DOI: 10.5223/pghn.2014.17.2.104
72 - Prabhasankar P, Ganesan P, Bhaskar N, Hirose A, Stephen N, Gowda LR, Hosokawa M, Miyashita K. Edible Japanese seaweed, wakame (Undaria pinnatifida) as an ingredient in pasta: Chemical, functional and structural evaluation. Food Chemistry. 2009;115:501-508. DOI: 10.1016/j.foodchem.2008.12.047
73 - Kadam SU, Prabhasankar P. Marine foods as functional ingredients in bakery and pasta products. Food Research International. 2010;43:1975-1980. DOI: 10.1016/j.foodres.2010.06.007
74 - Niranjana R, Gayathri R, Stephen NM, Sugawara T, Hirata T, Myashita K, Ganesan P. Carotenoids modulate the hallmarks of cancer cells. Journal of Functional Foods. 2015;18:968-985. DOI: 10.1016/j.jff.2014.10.017
75 - Jomova K, Valko M. Health protective effects of carotenoids and their interactions with other biological antioxidants. European Journal of Medicinal Chemistry. 2013;70:102-110. DOI: 10.1016/j.ejmech.2013.09.054
76 - Shahidi F, Ambigaipalan P. Novel functional food ingredients from marine sources. Current Opinion in Food Science. 2015;2:123-129. DOI: 10.1016/j.cofs.2014.12.009
77 - Lai CS, Wu JC, Pan MH. Molecular mechanism on functional food bioactives for anti-obesity. Current Opinion in Food Science. 2015;2:9-13. DOI: 10.1016/j.cofs.2014.11.008
78 - Revuelta JL, Buey RM, Ledesma-Amaro R, Vandamme EJ. Microbial biotechnology for the synthesis of (pro) vitamins, biopigments and antioxidants: Challenges and opportunities. Microbial Biotechnology. 2016;9:564-567. DOI: 10.1111/1751-7915.12379
79 - Curhan SG, Stankovic KM, Eavey RD, Wang M, Stampfer MJ, Curhan GC. Carotenoids, vitamin A, vitamin C, vitamin E, and folate and risk of self-reported hearing loss in women. American Journal Clinical Nutrition. 2015;102:1167-1175. DOI: 10.3945/ajcn.115.109314
80 - Bakan E, Akbulut ZT, Inanç AL. Carotenoids in foods and their effects on human health. Akademik Gıda. 2014;12:61-68.
81 - Lu CC, Yen GC. Antioxidative and anti-inflammatory activity of functional foods. Current Opinion in Food Science. 2015;2:1-8. DOI: 10.1016/j.cofs.2014.11.002
82 - Castro-Puyana M, Pérez-Sánchez A, Valdés A, Ibrahim OHM, Suarez-Álvarez S, Ferragut JA, Micol V, Cifuentes A, Ilbáñez E, García-Cañas V. Pressurized liquid extraction of Neochloris oleoabundans for the recovery of bioactive carotenoids with anti-proliferative activity. Food Research International, in press. DOI: 10.1016/j.foodres.2016.05.021
83 - Zhang X, Zhao W, Hu L, Zhao L, Huang J. Carotenoids inhibit proliferation and regulate expression of peroxisome proliferators-activated receptor gamma (PPARc) in K562 cancer cells. Archives of Biochemistry and Biophysics. 2011;512:96-106. DOI: 10.1016/
84 - Nishino H, Murakoshi M, Tokuda H, Satomi Y. Cancer prevention by carotenoids. Archives of Biochemistry and Biophysics. 2009;483:165-168. DOI: 10.1016/