Identification of a novel carotenoid, 2'-isopentenylsaproxanthin, by Jejuia pallidilutea strain 11shimoA1 and its increased production under alkaline condition.

Carotenoids are a class of naturally occurring pigment, carrying out important biological functions in photosynthesis and involved in environmental responses including nutrition in organisms. Saproxanthin and myxol, which have monocyclic carotenoids with a γ-carotene skeleton, have been reported to show a stronger antioxidant activity than those with ß-carotene and zeaxanthin. In this research, a yellow-orange bacterium of strain 11shimoA1 (JCM19538) was isolated from a seaweed collected at Nabeta Bay (Shizuoka, Japan). The 16S rRNA gene sequence of strain 11shimoA1 revealed more than 99.99 % similarity with those of Jejuia pallidilutea strains in the family Flavobacteriaceae. Strain 11shimoA1 synthesized two types of carotenoids. One of them was (3R, 3'R)-zeaxanthin with dicyclic structure and another was identified as (3R, 2'S)-2'-isopentenylsaproxanthin, a novel monocyclic carotenoid with pentenyl residue at C-2' position of saproxanthin, using FAB-MS, (1)H NMR, and CD analyses. Culturing strain 11shimoA1 in an alkaline medium at pH 9.2 resulted in a markedly increased in production of 2'-isopentenylsaproxanthin per dry cell weight, but a decreased in zeaxanthin production as compared to their respective production levels in medium with pH 7.0. These carotenoids are likely to play some roles in the adaptation of the bacterium to the environmental conditions.

Astaxanthin ameliorates heat stress-induced impairment of blastocyst development in vitro:--astaxanthin colocalization with and action on mitochondria--.

PURPOSE: The effects of astaxanthin (Ax) on the in vitro development of bovine embryos cultured under heat stress were investigated in combination with the assessment of its cellular accumulation and action on mitochondrial membrane potential (ΔΨm). METHODS: Bovine ≥8-cell embryos were collected on day 3 after in vitro fertilization and exposed to single (day 4) or repeated (day 4 and 5) heat stress (10 h/day at 40.5 °C). Ax was added into culture medium under the repeated heat stress and blastocyst development was evaluated. The cellular uptake of Ax in embryos was examined using bright-field and confocal laser-scanning microscopy, and high-performance liquid chromatography. The relationship between Ax and mitochondria localization was assessed using MitoTracker dye. The effects of Ax on ΔΨm were investigated using JC-1 dye. RESULTS: Blastocyst development in the repeated heat stress treatment decreased significantly (P < 0.05) compared with those in single heat stress or normal thermal treatment. The addition of Ax into culture medium did lead to a significant recovery in blastocyst development in the repeated heat-treated group. Ax was detected in cytoplasm of embryos and observed to colocalize with mitochondria. Ax recovered ΔΨm in embryos that was decreased by the heat treatment. CONCLUSIONS: Ax ameliorated the heat stress-induced impairment of blastocyst development. Our results suggest that the direct action of Ax on mitochondrial activity via cellular uptake is a mechanism of the ameliorating effects.

[Composition and content of carotenoids in body of the Black Sea gastropod mollusc Rapana venosa (Valenciennes, 1846)].

There were studied content and composition of carotenoids in body of the Black Sea gastropod mollusc Rapana venosa. To separate and identify this group of compounds, methods of thin layer and high performance liquid chromatography, mass-spectra and spectra in UV-VIS range were used. There were identified 84-87% of carotenoids. The main proportion belonged to mytiloxanthine--28-30%. The rest--pectenolon, diatoxanthin, alloxanthin--accounted for 10-15% of the total content except for beta-carotene--3-4%. The fraction is also isolated which contains complex esters of pectenolon, diatoxanthin, alloxanthin, and mytiloxanthin. It amounts to 17-20 % of pigments of carotenoid line. Minor components were not determined. Pathways of metabolic transformation of carotenoids in tissues of R. venosa are considered on the basis of study of the trophic system "mollusc-filtrator--mollusc-predator" by the example of Mytilus galloprovincialis and R. venosa.

Application of cDNA Macroarray for Simultaneous Detection of 12 Potato Viruses.

A complementary DNA (cDNA) macroarray was developed for simultaneous detection of 12 different potato viruses. A suitable region in the viral genome for each was selected for Alfalfa mosaic virus, Cucumber mosaic virus, Potato aucuba mosaic virus, Potato leafroll virus, Potato mop-top virus, Potato virus A, Potato virus M, Potato virus S, Potato virus X, Potato virus Y, Tomato ringspot virus, and Tomato spotted wilt virus, and their respective cDNAs were cloned into plasmid vectors. Capture probes for each virus ranging from 290 to 577 bp were generated by polymerase chain reaction (PCR) and immobilized on a nylon membrane. Total RNAs were extracted from each of these virus infected-plants, and cDNAs were synthesized from the RNA extracts using a random 9-mer primer. Subsequently, PCR reactions were performed using one primer pair for each of the 12 viruses. During PCR, amplified cDNAs were labeled with biotin and used as a target for hybridization analyses on a macroarray membrane. Hybridization signals between capture probes for the 12 viruses and their respective target cDNAs were observed using chemiluminescent or colorimetric detection. In all viruses, hybridization signals with capture probes were detected only when homologous virus targets were examined, and no hybridization to healthy plant extract was observed, facilitating identification of each virus. The results by colorimetric detection agreed with those obtained using chemiluminescence. The macroarray method developed was 5 × 102 to 4 × 106 times more sensitive than enzyme-linked immunosorbent assay and 5 to 5 × 104 times more sensitive than reverse-transcription PCR, except for Alfalfa mosaic virus. Colorimetric detection and substantial reduction in cross-hybridization signals much improved the method compared with other array-based detection methods for practical use.

Cancer chemopreventive activity of carotenoids in the fruits of red paprika Capsicum annuum L.

Capsanthin and related carotenoids isolated from the fruits of red paprika Capsicum annuum L. showed potent in vitro anti-tumor-promoting activity with inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). Among them, capsanthin diester and capsorbin diester showed strong inhibitory effects. Furthermore, capsanthin , capsanthin 3'-ester and capsanthin 3,3'-diester , major carotenoids in paprika, exhibited potent anti-tumor-promoting activity in an in vivo mouse skin two-stage carcinogenesis assay using 7, 12-dimethylbenz[a]anthracene as an initiator and TPA as a promoter.

Modifying effects of carotenoids on superoxide and nitric oxide generation from stimulated leukocytes.

Excessive and prolonged generation of superoxide (O2-) and nitric oxide (NO) from inflammatory leukocytes is associated with several lifestyle-related diseases, including cancer. In the present study, we screened 19 natural carotenoids for their modifying effects on O2- and NO generation from differentiated human promyelocytic HL-60 cells and mouse macrophage RAW 264.7 cells, respectively. Of the carotenoids tested, halocynthiaxanthin, isolated from oysters, showed the highest suppressive effect on the generation of both free radicals. The inhibitory potencies of certain carotenoids on radical generation markedly exceeded that of beta-carotene. In addition, some important structural moieties regulating radical generation suppression are discussed.

An antitumor promoter from Moringa oleifera Lam.

In the course of studies on the isolation of bioactive compounds from Philippine plants, the seeds of Moringa oleifera Lam. were examined and from the ethanol extract were isolated the new O-ethyl-4-(alpha-L-rhamnosyloxy)benzyl carbamate (1) together with seven known compounds, 4(alpha-L-rhamnosyloxy)-benzyl isothiocyanate (2), niazimicin (3), niazirin (4), beta-sitosterol (5), glycerol-1-(9-octadecanoate) (6), 3-O-(6'-O-oleoyl-beta-D-glucopyranosyl)-beta-sitosterol (7), and beta-sitosterol-3-O-beta-D-glucopyranoside (8). Four of the isolates (2, 3, 7, and 8), which were obtained in relatively good yields, were tested for their potential antitumor promoting activity using an in vitro assay which tested their inhibitory effects on Epstein-Barr virus-early antigen (EBV-EA) activation in Raji cells induced by the tumor promoter, 12-O-tetradecanoyl-phorbol-13-acetate (TPA). All the tested compounds showed inhibitory activity against EBV-EA activation, with compounds 2, 3 and 8 having shown very significant activities. Based on the in vitro results, niazimicin (3) was further subjected to in vivo test and found to have potent antitumor promoting activity in the two-stage carcinogenesis in mouse skin using 7,12-dimethylbenz(a)anthracene (DMBA) as initiator and TPA as tumor promoter. From these results, niazimicin (3) is proposed to be a potent chemo-preventive agent in chemical carcinogenesis.

Capsanthone 3,6-epoxide, a new carotenoid from the fruits of the red paprika Capsicum annuum L.

The structure of a new carotenoid, isolated from the fruits of the red tomato-shaped paprika Capsicum annuum L., was elucidated to be (3S,5R,6S,5'R)-3,6-epoxy-5,6-dihydro-5-hydroxy-beta,kappa-carotene-3',6'-dione by spectroscopic analyses, including fast atom bombardment collision-induced dissociation-mass spectrometry/mass spectrometry (FAB CID-MS/MS) and was designated capsanthone 3,6-epoxide. Capsanthone 3,6-epoxide is assumed to be an oxidative metabolite of capsanthin 3,6-epoxide in paprika.

Isolation of a series of apocarotenoids from the fruits of the red paprika Capsicum annuum L.

Eleven apocarotenoids (1-11) including five new compounds, 4, 6, 9, 10 and 11, were isolated from the fruits of the red paprika Capsicum annuum L. The structures of new apocarotenoids were determined to be apo-14'-zeaxanthinal (4), apo-13-zeaxanthinone (6), apo-12'-capsorubinal (9), apo-8'-capsorubinal (10), and 9,9'-diapo-10,9'-retro-carotene-9,9'-dione (11) by spectroscopic analysis. The other six known apocarotenoids were identified to be apo-8'-zeaxanthinal (1), apo-10'-zeaxanthinal (2), apo-12'-zeaxanthinal (3), apo-15-zeaxanthinal (5), apo-11-zeaxanthinal (7), and apo-9-zeaxanthinone (8) which have not been previously found in paprika. These apocarotenoids were assumed to be oxidative cleavage products of C(40) carotenoid such as capsanthin in paprika.

The simultaneous differentiation of Potato virus Y strains including the newly described strain PVY(NTN-NW) by multiplex PCR assay.

New recombinant strain and genotype of PVY, designated as PVY(NTN-NW) and SYR-III, respectively, shared properties with PVY(NTN) and PVY(N)W has been reported recently. PVY(NTN-NW) predominated in potato fields in Syria and was able to induce potato tuber necrotic ringspot disease (PTNRD). Due to the rapid spread of the recombinant strains of PVY which might be the case of PVY(NTN-NW), a specific and reliable detection method is an essential step to control this strain and minimize its spread. The shared properties of PVY(NTN-NW) and SYR-III with PVY(NTN) and PVY(N)W, however, complicate their identification involving multiple detection methods. Therefore, a multiplex polymerase chain reaction (PCR), that relies on a combination of previously published and newly designed primers was developed for the detection and identification of PVY(NTN-NW) and SYR-III in single or mixed infections with the main PVY strains, PVY(O), PVY(N), PVY(NTN) and PVY(N)W. In addition, the present PCR assay was able to detect the recombination points in the P1 region enabling the differentiation of the variable genotypes of the recombinant strains PVY(NTN-NW), PVY(NTN) and PVY(N)W. The reliability of this PCR assay was confirmed using a significant number of well characterized PVY isolates collected from Syria and Japan including those of PVY(NTN-NW), SYR-III, PVY(O), NA-PVY(N), PVY(N)W and PVY(NTN).

Phaeospirillum chandramohanii sp. nov., a phototrophic alphaproteobacterium with carotenoid glycosides.

A Gram-negative, spiral-shaped, phototrophic, purple non-sulfur bacterial strain, designated JA145T, was isolated from a freshwater habitat. Cells of strain JA145T were motile by means of a monopolar flagellum. Intracellular photosynthetic membranes were of the stacked type. Bacteriochlorophyll a and the carotenoid lycopene and its glucosides were present as photosynthetic pigments. There was no vitamin requirement for strain JA145T. The predominant cellular fatty acids were C16:1omega7c/C16:1omega6c (22.24%), C16:0 (22.97%) and C18:1omega7c (43.24%). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain JA145T clustered with species of the genus Phaeospirillum, in the class Alphaproteobacteria. The highest sequence similarities of strain JA145T were found with the type strains of Phaeospirillum fulvum (96.12%) and Phaeospirillum molischianum (96.19%). Based on the 16S rRNA gene sequence analysis and the morphological and physiological characteristics, strain JA145T is considered to represent a novel species, for which the name Phaeospirillum chandramohanii sp. nov. is proposed. The type strain is JA145T (=JCM 14933T=KCTC 5703T=NBRC 104961T).

Structures of carotenoids with 5,6-dihydro-beta-end groups from the spindle shell Fusinus perplexus.

A series of pirardixanthin derivatives, 1, 2, 3, 4, and 5, possessing the 5,6-dihydro-beta-end group were isolated from the spindle shell Fusinus perplexus.(1,2) Their structures and absolute configurations were determined by modern spectroscopic analysis to be (4S,5S,6S,4'S,5'S,6'S)-5,6,5',6'-tetrahydro-beta,beta-carotene-4,4'-diol (1), (3S,4R,5S,6S,4'S,5'S,6'S)-5,6,5',6'-tetrahydro-beta,beta-carotene-3,4,4'-triol (2), (3S,4R,5S,6S,3'S,4'R,5'S,6'S)-5,6,5',6'-tetrahydro-beta,beta-carotene-3,4,3',4'-tetrol (3), (5S,6S,4'S,5'S,6'S)-4'-hydroxy-5,6,5',6'-tetrahydro-beta,beta-caroten-4-one (4), and (4'S,5'S,6'S)-4'-hydroxy-5',6'-dihydro-beta,beta-caroten-4-one (5).

Myxoxanthophyll in Synechocystis sp. PCC 6803 is myxol 2'-dimethyl-fucoside, (3R,2'S)-myxol 2'-(2,4-di-O-methyl-alpha-L-fucoside), not rhamnoside.

We identified the molecular structures of the carotenoids in Synechocystis sp. PCC 6803. Myxoxanthophyll in this cyanobacterium was myxol 2'-dimethyl-fucoside, (3R,2'S)-myxol 2'-(2,4-di-O-methyl-alpha-L-fucoside). The sugar moiety of the pigment was not rhamnose but dimethylated fucose, which has not been reported in carotenoid glycosides. The other carotenoids were beta-carotene, (3R,3'R)-zeaxanthin, echinenone, (3'R)-3'-hydroxyechinenone and deoxymyxol 2'-dimethyl-fucoside, (2'S)-deoxymyxol 2'-(2,4-di-O-methyl-alpha-L-fucoside). Generally, the group of polar carotenoids in cyanobacteria is referred to as myxoxanthophyll, and the structure is considered to be myxol 2'-rhamnoside. Since the name myxoxanthophyll can not specify the sugar moiety and the identification of the sugar moiety is unfeasible in many cyanobacteria, we propose the following naming convention: when the sugar moiety is unknown, the name is myxol glycoside, when known, as in the case of rhamnose and alpha-L-fucose, they should be named myxol 2'-rhamnoside and myxol 2'-alpha-L-fucoside, respectively.

Salmoxanthin, deepoxysalmoxanthin, and 7,8-didehydrodeepoxysalmoxanthin from the Salmon Oncorhynchus keta.

The absolute configuration of salmoxanthin (5,6-epoxy-5,6-dihydro-beta,epsilon-carotene-3,3',6'-triol) (1) first isolated from the salmon Oncorhynchus keta was determined to be 3S, 5R, 6S, 3'S, 6'R. Furthermore, two minor carotenoids, deepoxysalmoxanthin (2) and 7,8-didehydrodeepoxysalmoxanthin (3), were isolated, and their structures were determined to be (3R,3'S,6'R)-beta,epsilon-carotene-3,3',6'-triol and (3R,3'S,6'R)-7,8-didehydro-beta,epsilon-carotene-3,3',6'-triol by (1)H NMR and CD spectroscopy.

Purification and structural determination of an inhibitor of starfish oocyte maturation from a Bacillus species.

Inhibitors of bacterial origins of starfish oocyte maturation were sought to obtain biologically active substances which act on either hormonal signal transduction or cell cycle regulation. An oocyte maturation-inhibiting substance found in culture fluid of a Bacillus species was purified to homogeneity. This substance possessed the nature of a detergent and specifically inhibited 1-methyladenine-induced oocyte maturation (50% inhibitory concentration, 3.3 microM) but not dithiothreitol-induced maturation. Its total structure was established to be the lactone of 3-hydroxy-13-methyltetradecanoyl-Glu-Leu-Leu-Val-Asp-Leu -Leu through COOH of the carboxy-terminal Leu. This structure is identical to surfactin, although although the configuration of the substance's amino acid residues has not yet been determined. Surfactin was shown to be identical with this substance in its inhibitory effect on starfish oocyte maturation as well as its chromatographic and electrophoretic properties. Therefore, it was concluded that the oocyte maturation-inhibiting substance produced by a Bacillus species is surfactin.

The first isolation of enantiomeric and meso-zeaxanthin in nature.

Racemic mixtures of (3RS, 3'RS)-zeaxanthin were separated into the three optical isomers, (3R, 3'R)-zeaxanthin (1), (3R,3'S;meso)-zeaxanthin (2) and (3S,3'S)-zeaxanthin (3), by converting to their corresponding dibenzoates and by using HPLC on an optical resolution column Sumipax OA-2000. According to this procedure, it has been shown that only (1) is isolated from higher plants, shellfish, starfish, sea squirt, sea cucumber and then examined; on the other hand (1), (2) and (3) are isolated from zeaxanthin fraction of shrimp, fish and turtle examined. This is the first isolation of enantiomeric and meso-zeaxanthin in nature.

Structures of five new carotenoids from the oyster Crassostrea gigas.

Five new minor carotenoids, 1-5, were isolated from the oyster Crassostrea gigas. The structure of 1 was determined to be (3S,5R,6R,6'S)-3,5,6'-trihydroxy-3'-oxo-6,7-didehydro-5,6-dihydro-10,11,20-trinor-beta,epsilon-caroten-19',11'-olide 3-acetate by detailed analyses of NMR and CD data. The structures of the other carotenoids, 2-5, were also determined in a similar manner. In the FAB-MS/MS of 2-4, having the 5-hydroxy-3,6-epoxy-5,6-dihydro-beta-carotene moiety, the characteristic product ions resulting from the sequential cleavage of C-C bonds in the polyene chain were observed.

Dihydroxylycopene diglucoside diesters: a novel class of carotenoids from the phototrophic purple sulfur bacteria Halorhodospira abdelmalekii and Halorhodospira halochloris.

The carotenoids in Halorhodospira abdelmalekii and Halorhodospira halochloris were analyzed by spectroscopic methods. The carotenoid composition of the two species was almost the same. Both species contained substantial amounts of unusual carotenoid glycoside fatty acid esters, which have been found for the first time in phototrophic purple bacteria. Methoxy-hydroxylycopene glucoside was a major component, and dihydroxylycopene diglucoside and dihydroxylycopene diglucoside diester were also found. Lycopene, rhodopin, and 3,4,3',4'-tetrahydrospirilloxanthin were present in very small amounts. Methoxy, glucosyl, and glucosyl ester groups were observed as substituents at the positions of the two original hydroxyl groups of dihydroxylycopene and made up approximately 20, 50, and 20%, respectively, of the total end groups (100%). The fatty acid components of the three carotenoid glucoside esters were the same (C12:0 and C14:1) and were rare in the cellular lipids of the two species.

Isolation of stereoisomeric epoxy carotenoids and new acetylenic carotenoid from the common freshwater goby Rhinogobius brunneus.

Stereoisomeric epoxy carotenoids with 3,5-cis configuration, diadinoxanthin B [(3S,5S,6R,3'R)-diadinoxanthin] (1) and antheraxanthin B [(3S,5S,6R,3'R)-antheraxanthin] (2), along with diadinoxanthin A [(3S,5R,6S,3'R)-diadinoxanthin] and antheraxanthin A [(3S,5R,6S,3'R)-antheraxanthin], were isolated from the common freshwater goby Rhinogobius brunneus. This is the first example in nature of 3,5-cis carotenoid epoxides. Furthermore, a new acetylenic triol carotenoid, gobiusxanthin (3), was obtained, and its structure was determined to be 7,8-didehydro-beta, epsilon-carotene-3,3', 6'-triol by chemical and spectral data.

Comparative biochemical studies of carotenoids in fishes--XXIX. Isolation of new luteins, lutein F and lutein G from marine fishes.

New luteins, lutein F [(3R,3'R,6'S)-beta,epsilon-carotene-3, 3'-diol] and lutein G [(3S,3'R,6'S)-beta,epsilon-carotene-3,3'-diol] have been isolated from marine fishes.