Dietary astaxanthin: an excellent carotenoid with multiple health benefits.

Astaxanthin is a carotenoid widely found in marine organisms and microorganisms. With extensive use in nutraceuticals, cosmetics, and animal feed, astaxanthin will have the largest share in the global market for carotenoids in the near future. Owing to its unique molecular features, astaxanthin has excellent antioxidant activity and holds promise for use in biochemical studies. This review focuses on the observed health benefits of dietary astaxanthin, as well as its underlying bioactivity mechanisms. Recent studies have increased our understanding of the role of isomerization and esterification in the structure-function relationship of dietary astaxanthin. Gut microbiota may involve the fate of astaxanthin during digestion and absorption; thus, further knowledge is needed to establish accurate recommendations for dietary intake of both healthy and special populations. Associated with the regulation of redox balance and multiple biological mechanisms, astaxanthin is proposed to affect oxidative stress, inflammation, cell death, and lipid metabolism in humans, thus exerting benefits for skin condition, eye health, cardiovascular system, neurological function, exercise performance, and immune response. Additionally, preclinical trials predict its potential effects such as intestinal flora regulation and anti-diabetic activity. Therefore, astaxanthin is worthy of further investigation for boosting human health, and wide applications in the food industry.

Assessment of the antioxidant activities of representative optical and geometric isomers of astaxanthin against singlet oxygen in solution by a spectroscopic approach.

Astaxanthin (AST) is a natural antioxidant and has been widely applied as a food supplement. While astaxanthin has many isomers, there are few studies comparing its physicochemical properties. In this work, we were concerned about their antioxidant activities against external oxidative stresses, and specifically, the singlet oxygen (1O2) quenching capacities of the representative optical and geometric isomers of astaxanthin were examined. Methylene blue (MB) was used as the photosensitizer to produce 1O2, and 1,3-diphenylisobenzofuran (DPBF) was used to probe 1O2. Our results showed that the 1O2 quenching capacities of the optical isomers, including 3S,3'S, 3R,3'S, and 3R,3'R all-trans-astaxanthin, are identical. In contrast, the 1O2 quenching capacity of cis-astaxanthin is higher than that of all-trans-astaxanthin. As such, this work provides an effective spectroscopic approach to assessing the antioxidant activities of various forms of astaxanthin against singlet oxygen, and demonstrates the remarkable difference among the geometric isomers.

A Method of Solubilizing and Concentrating Astaxanthin and Other Carotenoids.

The valuable marine carotenoid, astaxanthin, is used in supplements, medicines and cosmetics. In this study, crustacyanin, an astaxanthin-binding protein, was used to solubilize and concentrate astaxanthin. The recombinant crustacyanin of European lobster spontaneously formed an inclusion body when it was over-expressed in Escherichia coli. In this study, fusing the NusA-tag to the crustacyanin subunits made it possible to express in a soluble fraction and solubilize astaxanthin in aqueous solution. By cutting off the NusA-tag, the crustacyanin subunits generated the pure insoluble form, and captured and concentrated astaxanthin. Overall, the attaching and releasing NusA-tag method has the potential to supply solubilized carotenoids in aqueous solution and concentrated carotenoids, respectively.

Characterization of Astaxanthin Nanoemulsions Produced by Intense Fluid Shear through a Self-Throttling Nanometer Range Annular Orifice Valve-Based High-Pressure Homogenizer.

Stable, oil-in-water nanoemulsions containing astaxanthin (AsX) were produced by intense fluid shear forces resulting from pumping a coarse reagent emulsion through a self-throttling annular gap valve at 300 MPa. Compared to crude emulsions prepared by conventional homogenization, a size reduction of over two orders of magnitude was observed for AsX-encapsulated oil droplets following just one pass through the annular valve. In krill oil formulations, the mean hydrodynamic diameter of lipid particles was reduced to 60 nm after only two passes through the valve and reached a minimal size of 24 nm after eight passes. Repeated processing of samples through the valve progressively decreased lipid particle size, with an inflection in the rate of particle size reduction generally observed after 2-4 passes. Krill- and argan oil-based nanoemulsions were produced using an Ultra Shear Technology™ (UST™) approach and characterized in terms of their small particle size, low polydispersity, and stability.

Carotenoid Nostoxanthin Production by Sphingomonas sp. SG73 Isolated from Deep Sea Sediment.

Carotenoids are used commercially for dietary supplements, cosmetics, and pharmaceuticals because of their antioxidant activity. In this study, colored microorganisms were isolated from deep sea sediment that had been collected from Suruga Bay, Shizuoka, Japan. One strain was found to be a pure yellow carotenoid producer, and the strain was identified as Sphingomonas sp. (Proteobacteria) by 16S rRNA gene sequence analysis; members of this genus are commonly isolated from air, the human body, and marine environments. The carotenoid was identified as nostoxanthin ((2,3,2',3')-ß,ß-carotene-2,3,2',3'-tetrol) by mass spectrometry (MS), MS/MS, and ultraviolet-visible absorption spectroscopy (UV-Vis). Nostoxanthin is a poly-hydroxy yellow carotenoid isolated from some photosynthetic bacteria, including some species of Cyanobacteria. The strain Sphingomonas sp. SG73 produced highly pure nostoxanthin of approximately 97% (area%) of the total carotenoid production, and the strain was halophilic and tolerant to 1.5-fold higher salt concentration as compared with seawater. When grown in 1.8% artificial sea salt, nostoxanthin production increased by 2.5-fold as compared with production without artificial sea salt. These results indicate that Sphingomonas sp. SG73 is an efficient producer of nostoxanthin, and the strain is ideal for carotenoid production using marine water because of its compatibility with sea salt.

Development of astaxanthin-loaded layer-by-layer emulsions: physicochemical properties and improvement of LPS-induced neuroinflammation in mice.

Astaxanthin (AST) has been shown to have neuroprotective effects; however, its bioavailability in vivo is low due to its hydrophobic properties. In this study, lactoferrin (LF) was prepared by heat-treatment at different temperatures, and on this basis, a layer-by-layer self-assembly method was used to construct double-layer emulsions with LF as the inner layer and polysaccharide (beet pectin, BP or carboxymethyl chitosan, CMCS) as the outer layer. Then AST was encapsulated in the emulsions and their physiochemical properties and function were investigated. The results indicated that high temperature heated LF (95 °C) showed a more stable structure than the lower temperature one, and the exposed internal nonpolar groups of LF could give the emulsion an enhanced stability. The rheology results showed that compared with CMCS, the double-layer emulsion formed by BP had a higher viscosity. In addition, the 95 °C LF-AST-BP emulsion showed the best stability among all the bilayer emulsions. The best emulsion was then used as a model drug to investigate its effects on lipopolysaccharide (LPS)-induced neuroinflammation and learning-memory loss in C57BL/6J mice. Through animal behavioral experiments, it was found that dietary supplementation with the AST emulsion could effectively improve the brain cognitive and learning memory impairment caused by inflammation. Transmission electron microscopy, mRNA and western blotting results also illustrated that the AST emulsion could alleviate neuroinflammation caused by LPS. This study provides a feasible scheme for exploring an AST loaded system and may be suitable for food and drug applications.

Evaluation and improvement of storage stability of astaxanthin isomers in oils and fats.

Astaxanthin Z-isomers potentially have greater bioavailability and biological activity than (all-E)-astaxanthin. However, the stability of the Z-isomers is lower than the all-E-isomer, which is a serious problem affecting its practical use. In this study, we investigated the impacts of different suspension media (oils and fats) and additives on astaxanthin isomer stability and identified suitable ones for astaxanthin stabilization. The evaluations showed that several vegetable oils and antioxidants significantly improved astaxanthin isomer stability, e.g., when soybean and sunflower oils were used as the suspension medium, astaxanthin isomers were hardly degraded; however the total Z-isomer ratio decreased from ~80% to ~50% during 6-week storage at 30 °C. Moreover, it was revealed that (9Z)-astaxanthin showed higher stability than the 13Z- and 15Z-isomers. Hence, to maintain astaxanthin concentration and the Z-isomer ratio over long periods, it is important to use suitable suspension mediums and antioxidants, and select a Z-isomerization method that increases (9Z)-astaxanthin ratio.

Astaxanthin Z-isomer-rich diets enhance egg yolk pigmentation in laying hens compared to that in all-E-isomer-rich diets.

The effects of feeding diets containing astaxanthin with different Z-isomer ratios to laying hens on egg qualities, such as astaxanthin concentration in egg yolk and yolk color, were investigated. As the astaxanthin source, a natural microorganism Paracoccus carotinifaciens was used. Astaxanthin with different Z-isomer ratios was prepared by thermal treatment with different conditions and then added to the basal diet at a final astaxanthin concentration of 8 mg/kg. We found that, as the Z-isomer ratios of astaxanthin in the diet increased, the astaxanthin concentration in egg yolk and the yolk color fan score also increased significantly. Importantly, feeding a 50.6% Z-isomer ratio diet increased astaxanthin concentration in egg yolk by approximately fivefold and the color fan score by approximately 2 compared to that in hens fed an all-E-isomer-rich diet. Moreover, we showed that feeding Z-isomer-rich astaxanthin to laying hens increased plasma astaxanthin concentration by more than five times in comparison to that in hens fed an all-E-isomer-rich diet. These results indicate that Z-isomers of astaxanthin have higher bioavailability than that of the all-E-isomer and thus they exhibit greater egg yolk-accumulation efficiency.

Validation of a multi-analyte HPLC method for the determination of carotenoids used as feed additives in fish and poultry feed: results of an interlaboratory study.

The performance characteristics of a multi-analyte method for the determination of all 10 carotenoids authorised as feed additives within the EU were assessed via an interlaboratory study. The analytical method is based on reversed phase high performance liquid chromatography (RP-HPLC) coupled to an optical detector set at 410 nm. The analysis is particularly challenging due to the presence of various stereoisomers of each carotenoid, and the use of these compounds via natural or synthetic formulations, requiring a special sample preparation. EU regulations specifying the conditions of use set legal limits for these substances in compound feedingstuffs ranging from 6 mg kg-1 to 138 mg kg-1, depending on the individual carotenoid and the target animal for which the feed is supplemented with this carotenoid. The purpose of the multi-analyte method validated in this paper is to facilitate the monitoring of carotenoids at relevant levels when used as feed additives in compound feedingstuffs and pre-mixtures. The interlaboratory study delivered precision data for 43 different analyte/mass fraction/matrix combinations, covering a mass fraction range of the target analytes from 2.6 mg kg-1 to 3861 mg kg-1. The relative standard deviations for repeatability (RSDr) varied from 2.2 to 16.2 % with a mean value of 6 %, while the relative standard deviations for reproducibility (RSDR) varied from 6.8 to 39 % with a mean value of 21 %. Given the broad scope of the method covering 10 carotenoids added to compound feedingstuffs and pre-mixtures via different formulations, this multi-analyte method is considered fit for the intended purpose.

Hemin from porcine blood effectively stabilized color appearance and odor of prepared pork chops upon repeated freeze-thaw cycles.

This study was designed to evaluate changes in color following pork chop supplementation with porcine hemin, astaxanthin and paprika red in response to repeated freeze-thaw processes. Surface color analyses revealed that hemin significantly enhanced the appearance of the pork chops (P < 0.05), and the coloring efficiency of 0.10% hemin was similar to that of 0.20% astaxanthin and 0.08% paprika red. Sensory evaluations conducted on both raw and fried chops showed that hemin and astaxanthin significantly enhanced the overall acceptability of the chops before and after cooking. The color stability of the pork chops was also evaluated, and the results suggested that the hemin-colored chops were the most stable among the three, upon repeated freeze-thaw cycles. The electronic nose showed that the odor of the hemin-colored samples after 0, 3, and 7 freeze-thaw cycles was better than that of the other two groups. In conclusion, hemin may be a superior supplement for the large scale preparation of prepared pork chop.

Dietary Supplementation of Astaxanthin Improved the Growth Performance, Antioxidant Ability and Immune Response of Juvenile Largemouth Bass (Micropterus salmoides) Fed High-Fat Diet.

High-fat diet (HFD) usually induces oxidative stress and astaxanthin is regarded as an excellent anti-oxidant. An 8-week feeding trial was conducted to investigate the effects of dietary astaxanthin supplementation on growth performance, lipid metabolism, antioxidant ability, and immune response of juvenile largemouth bass (Micropterus salmoides) fed HFD. Four diets were formulated: the control diet (10.87% lipid, C), high-fat diet (18.08% lipid, HF), and HF diet supplemented with 75 and 150 mg kg-1 astaxanthin (HFA1 and HFA2, respectively). Dietary supplementation of astaxanthin improved the growth of fish fed HFD, also decreased hepatosomatic index and intraperitoneal fat ratio of fish fed HFD, while having no effect on body fat. Malondialdehyde content and superoxide dismutase activity were increased in fish fed HFD, astaxanthin supplementation in HFD decreased the oxidative stress of fish. The supplementation of astaxanthin in HFD also reduced the mRNA levels of Caspase 3, Caspase 9, BAD, and IL15. These results suggested that dietary astaxanthin supplementation in HFD improved the growth performance, antioxidant ability and immune response of largemouth bass.

Fucoxanthin activities motivate its nano/micro-encapsulation for food or nutraceutical application: a review.

Fucoxanthin is a xanthophyll carotenoid abundant in marine brown algae. The potential therapeutic effects of fucoxanthin on tumor intervention have been well documented, which have aroused great interests in utilizing fucoxanthin in functional foods and nutraceuticals. However, the utilization of fucoxanthin as a nutraceutical in food and nutrient supplements is currently limited due to its low water solubility, poor stability, and limited bioaccessibility. Nano/micro-encapsulation is a technology that can overcome these challenges. A systematic review on the recent progresses in nano/micro-delivery systems to encapsulate fucoxanthin in foods or nutraceuticals is warranted. This article starts with a brief introduction of fucoxanthin and the challenges of oral delivery of fucoxanthin. Nano/micro-encapsulation technology is then covered, including materials and strategies for constructing the delivery system. Finally, future prospective has been discussed on properly designed oral delivery systems of fucoxanthin for managing cancer. Natural edible materials such as whey protein, casein, zein, gelatin, and starch have been successfully utilized to fabricate lipid-based, gel-based, or emulsion-based delivery systems, molecular nanocomplexes, and biopolymer nanoparticles with the aid of advanced processing techniques, such as freeze-drying, high pressure homogenization, sonication, anti-solvent precipitation, coacervation, ion crosslinking, ionic gelation, emulsification, and enzymatic conjugation. These formulated nano/micro-capsules have proven to be effective in stabilizing and enhancing the bioaccessibility of fucoxanthin. This review will inspire a surge of multidisciplinary research in a broader community of foods and motivate material scientists and researchers to focus on nano/micro-encapsulated fucoxanthin in order to facilitate the commercialization of orally-deliverable tumor intervention products.

Engineering CrtW and CrtZ for improving biosynthesis of astaxanthin in Escherichia coli.

This study engineered ß-carotene ketolase CrtW and ß-carotene hydroxylase CrtZ to improve biosynthesis of astaxanthin in Escherichia coli. Firstly, crtW was randomly mutated to increase CrtW activities on conversion from ß-carotene to astaxanthin. A crtW* mutant with A6T, T105A and L239M mutations has improved 5.35-fold astaxanthin production compared with the wild-type control. Secondly, the expression levels of crtW* and crtZ on chromosomal were balanced by simultaneous modulation RBS regions of their genes using RBS library. The strain RBS54 selected from RBS library, directed the pathway exclusively towards the desired product astaxanthin as predominant carotenoid (99%). Lastly, the number of chromosomal copies of the balanced crtW-crtZ cassette from RBS54 was increased using a Cre-loxP based technique, and a strain with 30 copies of the crtW*-crtZ cassette was selected. This final strain DL-A008 had a 9.8-fold increase of astaxanthin production compared with the wild-type control. Fed-batch fermentation showed that DL-A008 produced astaxanthin as predominant carotenoid (99%) with a specific titer of 0.88 g·L-1 without addition of inducer. In conclusion, through constructing crtW mutation, balancing the expression levels between crtW* and crtZ, and increasing the copy number of the balanced crtW*-crtZ cassette, the activities of ß-carotene ketolase and ß-carotene hydroxylase were improved for conversion of ß-carotene to astaxanthin with higher efficiency. The series of conventional and novel metabolic engineering strategies were designed and applied to construct the astaxanthin hetero-producer strain of E. coli, possibly offering a general approach for the construction of stable hetero-producer strains for other natural products.

Docosahexaenoic acid-acylated astaxanthin ester exhibits superior performance over non-esterified astaxanthin in preventing behavioral deficits coupled with apoptosis in MPTP-induced mice with Parkinson's disease.

Non-esterified astaxanthin (AST) has been reported to exhibit protective effects from Parkinson's disease (PD). Notably, DHA-acylated astaxanthin ester (DHA-AST) is widely distributed in the seafood. However, whether DHA-AST has an effect on PD, and the differences between DHA-AST, non-esterified AST and the combination of non-esterified AST (AST) with DHA (DHA + AST) is unclear. In the present study, mice with PD, induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), were employed to investigate the effects of DHA-AST, AST and DHA + AST on Parkinson's disease. The rotarod test results showed that DHA-AST significantly suppressed the PD development in MPTP-induced mice, and was better than the effects of AST and DHA + AST. Further mechanistic studies indicated that all three astaxanthin supplements could inhibit oxidative stress in the brain. It was noted that DHA-AST had the best ability to suppress the apoptosis of dopaminergic neurons via the mitochondria-mediated pathway and JNK and P38 MAPK pathway in the brain among the three treated groups. DHA-AST was superior to AST in preventing behavioral deficits coupled with apoptosis rather than oxidative stress, and might provide a valuable reference for the prevention and treatment of neurodegenerative diseases.

Bioactive Polysaccharides from Seaweeds.

Bioactive compounds with diverse chemical structures play a significant role in disease prevention and maintenance of physiological functions. Due to the increase in industrial demand for new biosourced molecules, several types of biomasses are being exploited for the identification of bioactive metabolites and techno-functional biomolecules that are suitable for the subsequent uses in cosmetic, food and pharmaceutical fields. Among the various biomasses available, macroalgae are gaining popularity because of their potential nutraceutical and health benefits. Such health effects are delivered by specific diterpenes, pigments (fucoxanthin, phycocyanin, and carotenoids), bioactive peptides and polysaccharides. Abundant and recent studies have identified valuable biological activities of native algae polysaccharides, but also of their derivatives, including oligosaccharides and (bio)chemically modified polysaccharides. However, only a few of them can be industrially developed and open up new markets of active molecules, extracts or ingredients. In this respect, the health and nutraceutical claims associated with marine algal bioactive polysaccharides are summarized and comprehensively discussed in this review.

Nutraceutical characteristics of the brown seaweed carotenoid fucoxanthin.

Fucoxanthin (Fx), a major carotenoid found in brown seaweed, is known to show a unique and wide variety of biological activities. Upon absorption, Fx is metabolized to fucoxanthinol and amarouciaxanthin, and these metabolites mainly accumulate in visceral white adipose tissue (WAT). As seen in other carotenoids, Fx can quench singlet oxygen and scavenge a wide range of free radicals. The antioxidant activity is related to the neuroprotective, photoprotective, and hepatoprotective effects of Fx. Fx is also reported to show anti-cancer activity through the regulation of several biomolecules and signaling pathways that are involved in either cell cycle arrest, apoptosis, or metastasis suppression. Among the biological activities of Fx, anti-obesity is the most well-studied and most promising effect. This effect is primarily based on the upregulation of thermogenesis by uncoupling protein 1 expression and the increase in the metabolic rate induced by mitochondrial activation. In addition, Fx shows anti-diabetic effects by improving insulin resistance and promoting glucose utilization in skeletal muscle.

Isotonic Beverage Pigmented with Water-Dispersible Emulsion from Astaxanthin Oleoresin.

Astaxanthin is a powerful antioxidant, because it neutralizes free radicals and plays a vital role in the prevention of human diseases. The objective of this work was to develop an isotonic beverage (IB) of orange-red color, using an astaxanthin oleoresin emulsion (AOE) that is dispersible in water. This was carried out in order to simulate the color of commercial isotonic beverages (CIB) prepared from artificial pigments. The size of the AOE micelles ranged from 0.15 to 7.60 µm2. The color difference (ΔE) was similar for the samples exposed to dark as well as light conditions. The samples subjected to light stress showed pigment degradation after seven days, followed by a decrease in the concentration of astaxanthin; whereas, the samples exposed to dark conditions remained stable for seven days and then showed a decrease in the concentration of astaxanthin (this decrease ranged from 65% to 76% when compared to the initial content) after a period of 91 days. For the astaxanthin oleoresin (AO) and AOE, the oxygen radical absorbance capacity (ORAC) values reached 5224 and 1968 µmol of trolox equivalents (TE)/100 g, respectively. When exposed to light conditions, the addition of AOE in the IB led to its rapid degradation, while it remained stable in the samples exposed to the dark conditions.

A Quantitative Analysis Model Established to Determine the Concentration of Each Source in Mixed Astaxanthin from Different Sources.

Astaxanthin from different sources possesses different biological activities and optical isomers. The ingredients of astaxanthin mixtures from different sources on the market have often been mislabeled. Therefore, it is important to determine the sources of astaxanthin and their respective concentrations in a mixture. To solve this problem, a quantitative analysis model was established and further verified. The results showed that the deviation between the calculated concentration and the actual concentration ranged from 0 to 7 µg/mL, and the recovery rate was between 88.90% and 103.56%. This indicates that the quantitative analysis model of astaxanthin was feasible and reliable. This study not only has important applications in the astaxanthin mixture component determination but may also shed light on the quantitative analysis of other sample mixtures with stereoisomers from different sources.

Health benefits of fucoxanthin in the prevention of chronic diseases.

Fucoxanthin is a xanthophyll carotenoid abundant in macroalgae, such as brown seaweeds. When fucoxanthin is consumed, it can be esterified or hydrolyzed to fucoxanthinol in the gastrointestinal tract and further converted into amarouciaxanthin A in the liver. It has a unique chemical structure that confers its biological effects. Fucoxanthin has a strong antioxidant capacity by scavenging singlet molecular oxygen and free radicals. Also, it exerts an anti-inflammatory effect. Studies have demonstrated potential health benefits of fucoxanthin for the prevention of chronic diseases, such as cancer, obesity, diabetes mellitus, and liver disease. Animal studies have shown that fucoxanthin supplementation has no adverse effects. However, investigation of the safety of fucoxanthin consumption in humans is lacking. Clinical trials are required to assess the safety of fucoxanthin in conjunction with the study of mechanisms by which fucoxanthin exhibits its health benefits. This review focuses on current knowledge of metabolism and functions of fucoxanthin with its potential health benefits. This article is part of a Special Issue entitled Carotenoids recent advances in cell and molecular biology edited by Johannes von Lintig and Loredana Quadro.

Application of compressed fluid-based extraction and purification procedures to obtain astaxanthin-enriched extracts from Haematococcus pluvialis and characterization by comprehensive two-dimensional liquid chromatography coupled to mass spectrometry.

The green microalga Haematococcus pluvialis has been widely studied due to its capacity to accumulate great amounts of astaxanthin, a high-value carotenoid with biological activities. In the present work, two green compressed fluid-based processes, pressurized liquid extraction (PLE) and supercritical antisolvent fractionation (SAF), are integrated to obtain an astaxanthin-enriched extract from this microalga. PLE was carried out using pressurized ethanol as solvent, for 20 min, at 10 MPa, and 50 °C as extraction temperature. Subsequently, the obtained extract was processed by SAF to further purify the carotenoid fraction. The SAF process was optimized using a 3-level factorial experimental design and considering three experimental variables: (i) CO2 pressure (10-30 MPa), (ii) percentage of water in the PLE extract (20-50%), and (iii) PLE extract/supercritical-CO2 flow rate ratio (0.0125-0.05). Total carotenoid content was evaluated in both extracts and raffinates. Best results were obtained at 30 MPa, 0.05 feed/SC-CO2 mass flow rate, and 20% (v/v) of water in the feed solution, achieving values of 120.3 mg g-1 carotenoids in extract (in the SAF extract fraction), which were significantly higher than those obtained in the original PLE extract. In parallel, a new fast two-dimensional comprehensive liquid chromatography (LC×LC) method was optimized to get the full carotenoid profile of these extracts in less than 25 min. This is the first time that the use of a C30 column is reported in an on-line LC×LC system. Graphical abstract.