Carotenoids in fish IV. Salmonidae and Thymallidae from Polish waters

The author investigated the presence of various carotenoids in the Salmonidae and Thymallidae family by means of columnar and thin-layer chromatography. The investigations revealed the presence of the following carotenoids:

Abstract

- in the muscles of Salmo salar: astaxanthin (pure and ester), canthaxanthin, lutein and zeaxanthin.

- in the eggs of Salmo trutta m. trutta: β-carotene, iso- and zeaxanthin, lutein, taraxanthin and astaxanthin.

- in the eggs of Salmo trutta m. fario: β-carotene, canthaxanthin, 4-keto-4-hydroxy-β-carotene, astaxanthin (pure and ester), lutein, taraxanthin and astacene.

- in the eggs of Salmo gairdneri: β-carotene, γ-carotene (?), canthacanthin, isozeaxanthin, lutein and astaxanthin, and in the sperm Salmo gairdneri: β-carotene, γ-carotene (?), 4-keto-4-hydroxy-β-carotene, canthaxanthin, lutein and astaxanthin.

- in the eggs of Salvelinus fontinalis: ester astaxanthin, canthaxanthin, isozeaxanthin, lutein and astacene.

- in the eggs of Hucho hucho: β-carotene, tunaxanthin, lutein, taraxanthin and astaxanthin.

- in the eggs of Coregonus albula: β-carotene, 4-keto-4-hydroxy-β-carotene, ester astaxanthin, zeaxanthin, taraxanthin and astacene.

- in Coregonus lavaretus: a) in eggs: β-carotene, ester astaxanthin, canthaxanthin, iso- and zeaxanthin, lutein, taraxanthin and astacene b) in the sperm: canthaxanthin, 4-hydroxy-4-keto-β-carotene, isozeaxanthin and astaxanthin, and other organs: 4-hydroxy-α-carotene, canthaxanthin, tunaxanthin, monoepoxy lutein, lutein, iso- and zeaxanthin and astaxanthin.

- in the eggs of Coregonus peled: β-carotene, 4-keto-4-hydroxy-β-carotene, lutein, zeaxanthin, taraxanthin and astacene.

- in the eggs of Thymallus thymallus: β-carotene, tunaxanthin, lutein and astaxanthin.

     

Investigations of carotenoids in some animals of the Adriatic Sea V. Echinodermata

The author investigated the carotenoids in the Echinodermata from Adriatic sea by means of columnar and thin-layer chromatography. The following carotenoids were identified:

- in Coscinasterias tenuispina: β-carotene, isocryptoxanthin lutein, lutein-5, 6-epoxide, 4-hydroxy-4-keto-β-carotene, zeaxanthin, astaxanthin and asterinacid.

- in Marthasterias glacialis: β-carotene, echinenone, cryptoxanthin, lutein, lutein 5, 6-epoxide, 4-hydroxy-4-keto-β-carotene, zeaxanthin, astaxanthin ester, astaxanthin and 3, 4-didehydro-α-carotene.

- in Paracentrotus lividus: β-carotene, echinenone, cryptothin, isocryptoxanthin, lutein, lutein-5, 6-epoxide, 4-hydroxy-4-keto-β-carotene, zeaxanthin, astaxanthin, astaxanthin ester and asterinacid.

- in Sphaerechinus granularis: ,β-carotene, echinenone, cryptoxanthin, lutein, lutein-5, 6-epoxide, astaxanthin and guaraxanthin.

     

Comparative studies of carotenoids in the fauna of the Gullmar Fjord (Bohuslan,Sweden) III. Echinodermata

The author investigated the presence of various carotenoids in the Echinodermata from Gullmar Fjord (Bohuslan, Sweden) by means of columnar and thin-layer chromatography. The investigations revealed the presence of the following:

- inHenricia sanguinolenta:β-carotene, echinenone, canthaxanthin, guraxanthin, lutein-5, 6-epoxide and astaxanthin.

- inAmphiura filiformis: canthaxanthin, cryptoxanthin, lutein, lutein-5, 6-epoxide, isozeaxanthin, zeaxanthin, astaxanthin and 4-hydroxy-4-keto-β-carotene.

- inAmphipholis squamata:β-carotene, cryptoxanthin, lutein, lutein-5, 6-epoxide, astaxanthin, astaxanthin ester, asterin-acid and rubixanthin derivative.

- inOphiopholis aculeata: canthaxanthin, cryptoxanthin, isozeaxanthin, astaxanthin, astaxanthin ester, asterinacid, 4-hydroxy-4-keto-β-carotene, hydroxy rubixanthin and gazaniaxanthin-like substances.

- inOphiothrix fragilis: canthaxanthin, lutein-5, 6-epoxide, isozeaxanthin, astaxanthin, astaxanthin ester, 4-hydroxy-4-keto-β-carotene, and hydroxy rubixanthin.

- inAntedon petatus:canthaxanthin, guaraxanthin, isozeaxan-thin, zeaxanthin, astaxanthin, astaxanthin ester and 4-keto-4-ethoxy-β-carotene.

- inEchinocardium cordatum:β-carotene,γ-carotene, canthaxanthin, lutein, isozeaxanthin, zeaxanthin, astaxanthin and astaxanthin ester.

- inSpatangus purpureus: isozeaxanthin, astaxanthin, astaxanthin ester and 4-hydroxy-4-keto-β-carotene.

     

Precence of β-Carotene and Xantophylls in two species of the genus Niphargus (Crustacea: Amphipoda)

The presence of carotenoids in the Niphargus tatrensis Wrzesniowski and Niphargus aquilex schellenbergi Karaban has been investigated. In extracts separated by means of column and thin-layer chromatography, the following carotenoids were identified:

1. in Niphargus tatrensis: β-carotene, astaxanthin ester, rubixanthin, celaxanthin and astaxanthin.
2. in Niphargus aquilex schellenbergi: cantha-xanthin, astaxanthin ester, isozeaxanthin (only from Jeker), 4-keto-4′-hydroxy-β-carotene (only from Terzietebronbos), rubixanthin, celaxanthin and astaxanthin.

It was not possible to identifity the sexth fractions.     

Metabolism of carotenoids in sea-urchin Pseudocentrotus depressus

• 1.1. Feeding experiments with β,β-carotene, canthaxanthin and astaxanthin on the sea urchin Pseudocentrotus depressus were investigated.
• 2.2. In the case of β,β-carotene group, β-carotene was accumulated, β-isocryptoxanthin appeared and β-echinenone increased 6.8 times as much as the control group. On the other hand, in canthaxanthin and astaxanthin groups, canthaxanthin and astaxanthin increased significantly, respectively. The metabolic products of these carotenoids could not be found.
• 3.3. It was concluded that β,β-carotene was bioconverted to β-echinenone via β-isocryptoxanthin in P. depressus and could not be oxidatively metabolized beyond β-echinenone.

     

Characteristics of carotenoid features in muscle and ovary from anadromous and river resident types of Alaskan dolly varden charr (Salvelinus malma malma)

• 1.1. The carotenoids in the muscles and eggs from two types of natural Alaskan dolly varden charr, anadromous and river resident types, were examined.
• 2.2. In the muscle from the anadromous charr, astaxanthin was the major component comprising more than 70% of the total, followed by idoxanthin and 4-keto-zeaxanthin.
• 3.3. The egg carotenoid features in the river resident charr were more complicated compared with those in the anadromous fish. A considerable proportion of unidentified carotenoids was found in the eggs from the river resident charr.
• 4.4. Idoxanthin was the main component along with considerable propotions of β-carotene tetrol and astaxanthin in the eggs from the anadromous charr, whereas zeaxanthin and lutein were detected besides idoxanthin and β-carotene tetrol in the eggs from the river resident fish.
• 5.5. The prey was considered to be responsible for the difference in the carotenoid features of the eggs from the anadromous and river resident charr.

     

Presence of carotenoids in Gordius aquaticus (Nematomorpha,Nemathelminthes) specimens from deep wells

By means of columnar and thin-layer chromatography, the presence of carotenoids in Gordius aquaticus L. (Nematomorpha, Nemathelminthes) from deep wells was studied.The investigations revealed the presence of the following carotenoids: -carotene, mutatochrome, -cryptoxanthin, ,-carotene epoxide,lutein, zeaxanthin and astaxanthin ester.     

Carotenoids in fish. XXIV. Sardina pilchardus Walb. (Clupeidae)

The author investigated the presence of various carotenoids in Sardina pilchardus Walb. from the coast of Southern Europe.The presence of the following carotenoids has been stated: -carotene, -carotene epoxide, -cryptoxanthin, canthaxanthin, lutein epoxide, zeaxanthin, astaxanthin (free and ester form) and mutatochrome. The dominant carotenoid in all the parts of the body was astaxanthin, especially its ester form. The total content of carotenoid ranged from 10.537 (skin and muscles) to 116.309 µg/g fresh weight (liver).     

Genetic analysis of provitamin A carotenoid β-cryptoxanthin concentration and relationship with other carotenoids in maize grain (<Emphasis Type="Italic">Zea mays</Emphasis> L<Emphasis Type="Italic">.</Emphasis>)

Provitamin A (proVA) carotenoids are converted into retinol (vitamin A) in the human body, are the subject of human nutrition studies, and are targets for biofortification of staple crops. β-Carotene has been the principal target for enhancing levels of proVA. There is recent interest in enhancing the proVA carotenoid β-cryptoxanthin since it has excellent bioavailability, and in maize may be nearly as effective as β-carotene in providing retinol to humans. This study was designed to enhance our understanding of the genetic control of: levels of β-cryptoxanthin, conversion of β-carotene into β-cryptoxanthin and zeaxanthin, conversion of β-cryptoxanthin into zeaxanthin, and flux into and within the β-branch of carotenoid pathway. A biparental population derived from two inbreds with relatively high levels of β-cryptoxanthin and different ratios of β-carotene to β-cryptoxanthin and β-cryptoxanthin to zeaxanthin was studied. Three field replications of this F_2:3 population were grown, grain analyzed by liquid chromatography (LC), and composite interval mapping (CIM) performed to identify 90 quantitative trait loci (QTL) for carotenoids. We detected QTL for β-carotene/(β-cryptoxanthin + zeaxanthin) and (β-carotene + β-cryptoxanthin)/zeaxanthin ratios that contain candidate gene hydroxylase 4 (hyd4), which has not been previously associated with QTL for carotenoids in maize grain. Two color assessment methods, visual score and chromameter reading, were used to phenotype one replicate of the population for initial assessment as simple alternative measuring procedures. A common finding for LC and chromameter analysis included QTL on chromosome 5 that contain candidate gene lycopene β cyclase (lcyβ).     

The carotenoids of eggs of wild and farmed cod (Gadus morhua)

• 1.1. Eggs of wild cod, and of farmed cod fed (a) a diet supplemented with astaxanthin and (b) a diet supplemented with both astaxanthin and canthaxanthin, were analysed with respect to carotenoids.
• 2.2. The total carotenoid contents in eggs were 0.7 ppm for wild cod and 0.5 ppm for farmed cod.
• 3.3. Cod, having white flesh, deposit ketocarotenoids in the eggs, preferably astaxanthin.
• 4.4. Canthaxanthin can replace astaxanthin in the eggs, but astaxanthin appears to be deposited preferentially when both carotenoids are present in the diet.
• 5.5. The isomer distribution of (3S, 3′S):(3R, 3′S, meso):(3R, 3′R) astaxanthin in the eggs reflected the isomer composition of the diet.
• 6.6. Echinenone, 4′-hydroxyechinenone, adonixanthin and zeaxanthin encountered in cod eggs may represent reductive metabolites of canthaxanthin and astaxanthin.

     

Carotenoids in fish. XXVIII. Carotenoids in Micropterus salmoides (Lalépéde) Centrarchidae

The occurrence and contents of carotenoids in different body parts were investigated by column chromatography and TLC in Micropterus salmoides (Lalép).The following carotenoids were found: -carotene, -cryptoxanthin, -cryptoxanthin, echinenone, canthaxanthin, lutein, zeaxanthin, neothxanthin, tunaxanthin, -doradexanthin, -doradexanthin, idoxanthin, astaxanthin, astaxanthin ester, mutatochrome and mutatoxanthin.Their total contents varied within the range of 0.071–1.691 µg/g wet weight.     

Comparative biochemical studies of carotenoids in sea-urchins—II. The more primitive sea-urchins belonging to the orders Cidaroida,Echinothurioida, Diadematoida and Arbacioida

• 1.1. The carotenoids of seven species of more primitive sea-urchins, [orders Cidaroida (I), Echinothurioida (II), Diadematoida (III), and Arbacioida (IV)] were investigated from the comparative biochemical point of view.
• 2.2. β,β-carotene and β-echinenone have been isolated as major carotenoids in (I) and (III, IV), respectively. In (II), β,β-carotene, β-echinenone, canthaxanthin and (3S,3′S)-astaxanthin were foundto be predominant carotenoids.
• 3.3. The carotenoid patterns of (I) which is the most primitive sea-urchin from the phylogenetic point of view, and of (II) which is direct developers with non-feeding larvae, were quite different from those of the other sea-urchins showing typical development with feeding larvae.

     

Engineering of the carotenoid pathway in <Emphasis Type="Italic">Xanthophyllomyces dendrorhous</Emphasis> leading to the synthesis of zeaxanthin

Zeaxanthin is an essential nutrient for prevention of macular degeneration. However, it is limited in our diet. For the production of zeaxanthin, we have engineered zeaxanthin synthesis into a carotenoid mutant of Xanthophyllomyces dendrorhous which is blocked in astaxanthin synthesis and accumulates β-carotene instead. Two strategies were followed to reach high-yield zeaxanthin synthesis. Total carotenoid synthesis was increased by over-expression of genes HMGR, crtE, and crtYB encoding for limiting enzymes in the pathway leading to and into carotenoid biosynthesis. Then bacterial genes crtZ were used to extend the pathway from β-carotene to zeaxanthin in this mutant. The increase of total carotenoids and the formation of zeaxanthin is dependent on the number of gene copies of crtYB and crtZ integrated into the X. dendrorhous upon transformation. The highest zeaxanthin content around 500 μg/g dw was reached by shaking flask cultures after codon optimization of crtZ for Xanthophyllomyces. Stabilization of carotenoid and zeaxanthin formation in the final transformant in the absence of selection agents was achieved after passing through a sexual cycle and germination of basidiospores. The values for the transformant before and after stabilization were very similar resembling about 70 % of total carotenoids and corresponding to a conversion rate of 80 % for hydroxylation of β-carotene to zeaxanthin. The stabilized transformant allowed experimental small-scale fermentation yielding X. dendrorhous cells with a zeaxanthin content similar to the shaking flask cultures. Our result demonstrates the potential of X. dendrorhous for its development as a zeaxanthin producer and its suitability for large-scale fermentation.     

Investigations of carotenoids in some faunal elements of the Adriatic Sea. IV-molluscs

The carotenoids in the molluscsClanculus cruciatus, Patella coerulea, Mytilus galloprovincialis, Sepia officinalis andLoligo vulgaris from the Adriatic sea were investigated. Their presence was determined by means of columnar and thin-layer chromatography. The following carotenoids were found inC. cruciatus; mytiloxanthin-like, lutein, lutein ester, zeaxanthin and astaxanthin-like; inP. coerulea: mytiloxanthin-like, lutein, lutein ester, lutein-5,6-epoxide, zeaxanthin and astaxanthin-like; inM. galloprovincialis: -carotene, mytiloxanthin-like, lutein, lutein ester, lutein-5,6-epoxide and zeaxanthin; inS. officinalis: -carotene, lutein, lutein ester, tunaxanthin and zeaxanthin; inL. vulgaris: -carotene, -carotene, -carotene, -cryptoxanthin, isocryptoxanthin, isorenieratene, capsanthin, capsorubin, mutatochrome, triophaxanthin, zeaxanthin, 4-hydroxy--carotene and 4-keto--carotene     

Bioconversion pathway of astaxanthin into retinol2 in mature rainbow trout (Salmo Gairdneri Rich.)

• 1.1. Carotenoid and retinoid forms were analysed by HPLC in various tissues of mature rainbow trout fed for 11 months (7–9°C) with astaxanthin (50 and 100 mg/kg diet) or canthaxanthin (100 mg/kg diet).
• 2.2. Decreasing concentrations of canthaxanthin, echinenone and β-carotene, but no retinol₁, were found in the liver and skin of canthaxanthin-fed fish.
• 3.3. Higher retinol₂ concentrations were found in ovaries and testes of astaxanthin-fed fish compared to canthaxanthin and control groups.
• 4.4. A new metabolic pathway for direct conversion of astaxanthin into retinol₂ in gonads is proposed.

     

Carotenoids in fish XIV. The carotenoid content in the flesh of certain species from the Antarctic.

The authors investigated the presence of various carotenoids in the fish of certain species in the Rajidae (Raja georgiana), Muraenolepidae (Muraenolepis microps), Notothenidae rDissostichus eleginoides, Notothenia gibberifrons, Notothenid rossi-marmorata, Trematomus hansoni) and Chaenichthyidae (Chaenocephalus aceratus, Champsocephalus gunnari, Pseudochaenichthys georgianus) family from the Antarctic by means of columnar and thin-layer chromatography.The following carotenoids were identified: ß-carotene, -cryptoxanthin, canthaxanthin, flavoxanthin, isozeaxanthin, zeaxanthin, tunaxanthin, lutein-5, 6-epoxide, aurochrome, aurochrome-like, auroxanthin, astaxanthin, astaxanthin ester and 4-hydroxy--carotene.The total carotenoid content of this fishes was from 0.066 to 0.122 µg/g fresh weight.     

Change of carotenoid composition in crabs during embryogenesis

Changes of the qualitative and quantitative compositions of carotenoids are studied at various development stages of the external hard roe, determined based on color differences, for the species C. opilio, P. camtschaticus, and P. platypus. It has been revealed that the major carotenoids of the new egg are astaxanthin and β-carotene. Intermediate products of transformation of β-carotene into astaxanthin are identified: echinenone, canthaxanthine, and phenicoxanthine. The carotenoid content per embryo for the new hard roe of C. opilio (the orange egg) amounted to 22.7 ng, of P. camtschaticus and P. platypus (the violet egg)—to 49.2 and 23.3 ng, respectively. In the hard roe at the later development stage (the brown egg) the carotenoid content was decreased to 13.1 ng in C. opilio and to 20.1 ng in P. camtschaticus. Development of embryos is accompanied by accumulation of esterified carotenoids and a decrease of β-carotene and astaxanthine concentrations in all studied species.     

Investigation of seasonal variations in cow serum retinol and β -carotene by high performance liquid chromatographic method

• 1.1. High performance liquid chromatography (HPLC) was used to determine β-carotene and retinol in cow serum.
• 2.2. Two groups of state and private farm cows (Groups 1 and 2) were used to assess seasonal variation when different food sources were fed to cows on serum β -carotene and retinol concentrations.
• 3.3. Mean serum concentrations of β-carotene and retinol from October to April in both Groups 1 and 2 cows were lower (P < 0.05) than in the other months when the cows were fed various combination of maize silage, alfalfa and carrot residues and grass hay, respectively.
• 4.4. Mean serum β-carotene and retinol concentrations in June and July were higher (P < 0.05) than in other months when the cows were in pasture.
• 5.5. Mean serum β-carotene and retinol concentrations in May, August and September were lower (P < 0.05) than in June and July and higher (P < 0.05) than in other months when a lesser amount of green pasture was available to the cows.
• 6.6. There was a seasonal variation (P < 0.05) in serum β -carotene and retinol concentrations. When the carotene intake is very high, conversion of β -carotene to retinol decreases. Mean monthly serum β -carotene and retinol concentrations showed that combination of alfalfa hay and maize silage, and grass hay and carrot residues can maintain adequate serum β-carotene and retinol concentrations during the dry season.

     

Carotenoids in Ostrea edulis L. (Bivalvia: Ostreacea) from the lagoon of Venice (Italy)

By means of columnar and thin-layer chromatography the presence of carotenoids in Ostrea edulis L. (Bivalvia: Ostreacea) from the Lagoon Venice (Italy), was studied.The following carotenoids were founds: -, -, -carotene, -carotene epoxide, lutein (free and epoxide from), zeaxanthin, diatoxanthin, mutatoxanthin and astaxanthin. The dominating carotenoids were lutein.The total contents varied within the range of 5.4–8.1 µg/g wet weight.     

Molecular and spectroscopic characterisation of a low molecular weight seed storage protein from yellow mustard (Sinapis alba L.)

• 1.1. A 1.7S protein has been purified from mustard seeds (Sinapis alba L.). This protein, soluble in water and dilute salt solutions, is considered as an albumin and constitutes about 10% of the total soluble protein in mustard seeds.
• 2.2. Its molecular weight is approximately 15,000 and is composed of two polypeptide chains (M_r = 9500 and 5000), linked by two disulfide bridges.
• 3.3. The amino acid compositions of both subunits as well as of the native protein are reported, showing a strong homology with napins from Brassica napus L.
• 4.4. The ultraviolet absorption, fluorescence emission and circular dichroism spectra of the purified protein have been obtained. The mustard protein exhibits about 50% α-helix with a very low β-structure content. Based on its structural characteristics, a zein-like packing is proposed for this protein from mustard seeds.