Carotenoids of Rowan Berries

The berries of Sorbus aucuparia have been investigated for theircarotenoid contents. The pigments identified were: phytofluene,-, ß-carotene, cryptoxanthin, monoepoxy--carotene,monoepoxy-ß-carotene, aurochrome, and mutatochrome.Usually when green fruits ripen the control of carotenoid synthesisis removed when the chlorophylls disappear, there is a rapidincrease of carotenoids in an over-all oxidative manner anddifferent carotenoids appear. However, the results obtainedsuggest that a different mechanism takes place in S. aucuparia.This may be the exception that proves the rule.     

Carotenoid Pigment Changes in Ripening Momordica charantia Fruits

The carotenoid composition of Momordica charantia fruit (pericarp)at four levels of maturity was extensively investigated. Thenumber of carotenoids isolated increased from five in the immaturefruit to six at the mature-green and 14 at the partly-ripe andripe stages. Cryptoxanthin, which could not be isolated at theimmature and mature stages, accumulated rapidly at the onsetof ripening to become the principal pigment of the ripe fruit.Moderate increases were seen in ß-carotene, zeaxanthinand lycopene concentrations as ripening progressed. The reversetrend was observed with lutein and -carotene which were themajor pigments of the immature fruit. Prior to the colour break,only the hydroxy derivatives of -carotene (zeinoxanthin andlutein) could be detected; the ß-hydroxy compounds(cryptoxanthin and zeaxanthin) appeared and predominated thereafter.The hydroxy carotenoids of the ripe fruit were almost entirelyesterified in contrast to those of the unripe fruit which weremainly unesterified. Traces of flavochrome, 5, 6-monoepoxy-ß-carotene,mutatochrome, -carotene, -carotene, -carotene and rubixanthinwere detected in the partly-ripe and ripe fruits but not inthe immature and mature-green samples. Phytofluene was observedin trace levels at all stages.     

Observations on pigments and morphology of Gyrodinium aureolum Hulburt, a marine dinoflagellate containing 19' -hexanoyloxyfucoxanthin as the main carotenoid

Gyrodinium aureolum, a common "red tide" dinoflagellate in Europeanwaters often associated with fish mortality, was isolated fromthe Oslofjord, Norway, and analysed for chlorophylls and carotenoids.Besides chlorophyll a and c the following carotenoids were characterizedby thin-layer chromatography, visible light spectrophotometryand mass spectrometry: ß,-carotene, ß,ß-carotene,djatoxanthin, diadinoxanthin, 19'-hexanoyloxyfucoxanthin and3 xanthophylls which could not be correlated with hitherto structurallyknown carotenoids from dinoflagellates. G. aureolum deviatesfrom most dinoflagellates by the lack of peridinin, but showsaffinity with Gyrodinium sp.-A by the possession of 19'-hexanoyloxyfucoxanthin. Preliminary light microscopical observations on the internalstructure indicate that G. aureolum is uni-nucleate with a typicaldinokaryotic nucleus containing continually condensed chromosomes.The chloroplasts seem to possess an internal pyrenoid like someother dinoflagellates with deviating carotenoid pigmentation.The similarity in carotenoid pigmentation and chloroplast structureof Emiliania huxleyi (Prymnesiophyceae) and Gyrodinium sp.-Aand G.aureolum (Dinophyceae) is pointed out. The potential chemotaxonomicvalue of the carotenoid composition in establishing identitywith morphologically similar and ichthyotoxic dinoflagellatesis briefly discussed.     

The Gamete Pigments of Hormosira banksii (Phaeophyta)

The gamete pigments of Hormosira banksii have been separatedby thin-layer chromatography and identified by absorption spectrophotometry.Male gametes contain predominantly ß-carotene, andfemale gametes predominantly chlorophyll and fucoxanthin. Severalminor pigments have also been tentatively identified.     

A Novel Carotenoid Ester, Loroxanthin Dodecenoate, from Pyramimonas parkeae (Prasinophyceae) and a Chlorarachniophycean Alga

A novel carotenoid ester, which had previously been assumedtentatively and without full supporting data to be loroxanthin19-dodecenoate (Kohata and Watanabe 1989), was isolated andpurified from cultured strains of Pyramimonas parkeae (Prasinophyceae)and a chlorarachniophycean alga. From spectroscopic and chemicalevidence, including results of analysis by ¹H-NMR, FD-MS, GLCand CD, the compound was clearly identified as loroxanthin dodecenoate,(3R,3'R,6'R)-ß,-carotene-3,19,3'-triol 19-(2-trans-dodecenoate).A double bond of the dodecenoate was located at the 12 positionand was in the trans form, as is the case for that in a siphonaxanthinester. However, loroxanthin itself was absent from these algae.Other algal pigments identified were Chls a and b, ß-carotene,lutein A, zeaxanthin, violaxanthin and neoxanthin. ³ Present address: Nippon Roche Research Center, Kajiwara 200,Kamakura, Kanagawa, 247 Japan.     

Changes in Carotenoid Composition of Certain Roses with Age

The carotenoid composition of the anthers and styles and thefruits of four roses of different ages have been investigated.ß-carotene and its epoxides were present at all stagesstudied, as were the ß-carotene derived xanthophyllauroxanthin and the -carotene derived flavoxanthin and chrysanthemaxanthin.As the various parts aged, the control of carotenoid synthesiswas removed, oxidative processes took place with the resultthat very few members of the -carotene series were found, andepoxy-carotenoids and their derivatives were the main carotenoidspresent. Rubixanthin, 3-hydroxy--carotene characteristic ofrose hips was found in fairly large amounts in anthers and stylesand -carotene was probably the precursor of this pigment. Rosehips were also investigated for their vitamin-A contents whichwere not as high as those of All Gold flowers.     

Pigment synthesis in Cucurbita moschata cotyledons as influenced by CPTA and several inhibitors

2-(4-Chlorophenylthio) triethylamine hydrochloride (CPTA) inducedthe accumulation of a number of carotenes in pumpkin (Cucurbitamoschata) cotyledons, a tissue which does not normally accumulatethese pigments. Lycopene accumulated concomitantly with a decreasein -carotene and ß-carotene. CPTA appeared to inhibitcyclases involved in the synthesis of -carotene and ß-caroteneand to stimulate enzymes involved in lycopene synthesis. Cycloheximidereduced this CPTA induced enzyme synthesis while diphenylaminereduced CPTA induced carotene accumulation. Actinomycin D alonereduced accumulation of carotenes, but it did not affect CPTAinduced carotene accumulation. Rather lutein, violoxanthin andneoxanthin decreased and -carotene and ß-caroteneaccumulated. ¹Present address: Morioka Branch, Vegetable and Ornamental CropsResearch Station, Ministry of Agriculture and Forestry, Shimokuriyagawa,Morioka, Japan. (Received August 12, 1974; )     

Effect of Inhibitors on the Formation of Stereoisomers in the Biosynthesis of {beta}-Carotene in Dunaliella bardawil

An assay system was developed in which the effects of inhibitorsof ß-carotene biosynthesis in Dunaliella bardawilcould be tested. Since D. bardawil can be induced to accumulateover 10% of its dry weight as ß-carotene, it is particularlysuitable for such studies. Norflurazon a desaturation inhibitor,caused the accumulation of phytoene, or of phytoene and phytofluene,depending on the concentration employed. J-334, a substituted6-methylpyrimidine which also inhibits desaturation, causedthe accumulation of ß-zeacarotene, -carotene and phytoenein different proportions, depending on the concentration employed.The cyclization inhibitors, nicotine, CPTA and MPTA, severelyaffected the growth and survival of the alga, and their effectscould therefore not be studied directly. However, their actionwas observed indirectly by following the transformation of phytoenein norflurazon-pretreated phytoene-rich algae. Under these conditions,presence of the cyclase inhibitors caused the transformationof phytoene to lycopene, rather than to ß-carotene.The accumulated ß-carotene or the intermediates ß-zeacarotene,lycopene, -carotene, phytofluene and phytoene in D. bardawilwere all composed of two stereoisomers, tentatively assignedas the all-trans stereoisomer (55%) and the 9-cis stereoisomer(45%). This suggests that the isomerization reaction which leadsto the production of the presumed 9-cis isomers occurs earlyin the pathway of carotene biosynthesis, at or before phytoene,with no isomerization during the further transformations leadingto ß-carotene. (Received January 29, 1990; Accepted May 9, 1990)     

Comparative Studies of Acid Glycosidases from Three Legumes

The major isoenzymes of -mannosidase (EC 3.2.1.24 [EC] ) and ß-galactosidase(ECf 3.2.1.23 [EC] ) have been separated from cotyledons of gardenpea, Pisum sativum L. (Vicieae), chick pea, Cicer arietinumL. (Cicereae), and cowpea, Vigna unguiculata (L.) Walp. (Phaseoleae).Some of their properties have been determined, including pHoptima, K_m values for p-nitrophenyl glycosidc substrates, andthe effects of several inhibitors. Swainsonine, an indolizidinealkaloid, was the most effective inhibitor of mannosidase 1,with I₃₀ values of 5.6 x 10^–8 M (cowpea), 1x 10^–7M (chick pea) and 2.9 x 19^–7 M (pea). The most effectiveinhibitor of ß-galactosidase 2 from all sources wasD-galactonic acid-1,4-lactonwe (-lactone), with K_i values rangingbetween 3.0 and 3.9x 10^–3 M. An inhibitor of the E. coliß-galactosidose, p-aminophenyl thio-ß-D-galactopyranoside,did not inhibit any of the legume ß-galctosidases;rather it enhanced the activites of the enzymes from chick peaand cowpea cotyledons. Etiolated hull and seed tissues frompea pods developing in darkness contained similar acid glycosidaseactivities to normal green tissues, thus the chloroplast isan unlikely location for ß-galactosidase 2. The majorß-galactosidasesdetected with an indigogenic substrate (5-bromo-4-chloro-3-indoxyl-ß-D-galactopyranoside)following gel electrophoresis of extracts from pea hull, seedcoats and cotyledons appeared to be different from ß-galactosidase2. Acid glycosidase, cotyledon, isoenzyme, -lactone, legume, swainsonine     

The in vivo cleavage of carotenoids into retinoids in Chlamydomonas reinhardtii

A variety of carotenoids have been incorporated in vivo intoa carotenoid mutant of the unicellular alga, Chlamydomonas reinhardtii.The mutant can synthesize retinal from exogenous carotenoids.In this way the first step in biosynthesis of retinoids, namelyhow ß-carotene is converted into retinal, has beenstudied. Since retinal forms with opsin the photoreceptor forphototaxis in Chlamydomonas, the action-spectral peaks of thephototaxis restored by carotenoid incorporation were used topredict the products formed by the enzyme that cleaves thesecarotenoids. The data suggest that the physiologically relevantcleavage of ß-carotene into retinal is central ratherthan excentric. When apocarotenoids are substrates, the enzymetargets the double bond located a constant distance away fromthe carbonyl group on the acyclic end and, consequently, retinalis not produced. Interestingly, dehydro-analogues containinga triple bond are preferentially cleaved and oxidized at thetriple bond relative to the corresponding analogue with onlydouble bonds. Key words: Chlamydomonas reinhardtii, Chlorophyceae, retinoid biosynthesis, carotenoid cleavage, action spectrum     

Anthocyanin, Carotenoid, and Chlorophyll Changes in the Peel of Cox's Orange Pippin Apples during Ripening on and off the Tree

The concentrations of various peel pigments of Cox’s OrangePippin apples have been measured during ripening on the treeand during storage at 12 °C. Total chlorophyll decreased and total carotenoid increased atthe time of the respiration climacteric. These changes weremore pronounced in fruit maturing on the tree where a significantincrease of anthocyanin occurred; it did not occur in storedfruit. There was no consistent or marked difference in the ratesof destruction of chlorophylls a and b. The carotenoids found in the unripe fruit were those characteristicof photosynthetic tissue, ß-carotene, lutein, violaxanthin,and neoxanthin. These decreased to a greater or lesser extent,and at different rates, on and off the tree. Other carotenoidswhich increased greatly during ripening were identified as esters,mainly of violaxanthin. During the climacteric there is a transition from an assemblageof pigments associated with the chloroplast to that typicalof a chromoplast.     

Natural occurrence of phycoerythrocyanin-like pigment in cyanobacterial blooming samples dominated by Microcystis in Lake Kasumigaura

We describe the occurrence of phycocyanin and phycoerythrocyanin-likepigment in natural Microcystis colonies collected in Lake Kasumigaura.For differentiating pigments, the combination of simultaneousdetermination of molecular weights of and ß and subunitsby electrospray ionization mass spectrometry with HPLC separationand fluorescence measurements was used. It was suggested thatthe natural Microcystis colonies may contain the phycoerythrocyanin-likepigment, judged from the excitation and emission spectra. Measuredpigments consisted of different molecular weights of and ßsubunits, but the molecular weights of and ß subunitsin both phycocyanin and phycoerythrocyanin-like pigments werenot always identical.     

Changes in the Localization of {beta}-Glycerophosphatase Activity During the Infection of Phaseolus vulgaris by Colletotrichum lindemuthianum

Histochemical methods showed that epidermal cells of Phaseolusvulgaris (cv. Kievit) contained lysosomelike particles richin ß-glyceTophosphatase. The behaviour of these organellesduring infection by different physiological races of Colletotrichumlindemuthianum has been examined. Host cell death during theresistant (hypersensitive) response of the bean to infectionby incompatible races ß and occurred during the secondand third days after inoculation. Cell death appeared to becoincident with the release of ß-glycerophosphataseinto the cytoplasm and a reduction in size and number of stainingparticles. Invading incompatible hyphae were restricted to singlenecrotic cells. In contrast, for 4 days, infection by the compatiblerace caused little alteration in particulate staining whileconsiderable fungal colonization took place. Subsequent observationsrevealed a decrease in the number of enzyme-rich particles whichwas not associated with the appearance of diffuse staining evenafter cell necrosis. It is suggested that the release of ß-glycerophosphataseand possibly other hydrolases from the lysosome-like particlesof the host caused hypersensitive cell death, and that necrosiswas not controlled by the plant in this way during the susceptibleresponse.     

Ethylene regulation of carotenoid accumulation and carotenogenic gene expression in colour-contrasted apricot varieties (Prunus armeniaca)

In order to elucidate the regulation mechanisms of carotenoidbiosynthesis in apricot fruit (Prunus armeniaca), carotenoidcontent and carotenogenic gene expression were analysed as afunction of ethylene production in two colour-contrasted apricotvarieties. Fruits from Goldrich (GO) were orange, while Moniqui(MO) fruits were white. Biochemical analysis showed that GOaccumulated precursors of the uncoloured carotenoids, phytoeneand phytofluene, and the coloured carotenoid, ß-carotene,while Moniqui (MO) fruits only accumulated phytoene and phytofluenebut no ß-carotene. Physiological analysis showed thatethylene production was clearly weaker in GO than in MO. Carotenogenicgene expression (Psy-1, Pds, and Zds) and carotenoid accumulationwere measured with respect to ethylene production which is initiatedin mature green fruits at the onset of the climacteric stageor following exo-ethylene or ethylene-receptor inhibitor (1-MCP)treatments. Results showed (i) systematically stronger expressionof carotenogenic genes in white than in orange fruits, evenfor the Zds gene involved in ß-carotene synthesisthat is undetectable in MO fruits, (ii) ethylene-induction ofPsy-1 and Pds gene expression and the corresponding productaccumulation, (iii) Zds gene expression and ß-caroteneproduction independent of ethylene. The different results obtainedat physiological, biochemical, and molecular levels revealedthe complex regulation of carotenoid biosynthesis in apricotsand led to suggestions regarding some possible ways to regulateit. Key words: Apricot, carotenoid, ethylene, fruit, 1-MCP, Prunus armeniaca, ripening-related genes     

Isolation and Characterization of the Cytoplasmic Membranes from the Blue-Green Alga (Cyanobacterium) Anacystis nidulans

Cells of the blue-green alga (cyanobacterium) Anacyslis nidulanswere disintegrated, and their thylakoid membranes and cytoplasmicmembranes were isolated by floatation centrifugation on a sucrosedensity gradient. Electron micrographs revealed that the cytoplasmicmembranes formed single closed vesicles having diameters of200–400 nm. These membranes contained xanthophylls asthe major constituent pigments and rß-carotene andchlorophyll a as very minor ones. The major peaks in their absorptionspectra were due to carotenoids at 435, 455 and 487 nm, witha minor one due to chlorophyll a at 673 nm. These findings areconsistent with the yellow color of the cytoplasmic membranes.The absorption spectrum of the membranes in the carotenoid regionwas markedly affected by temperature: with a decrease in temperature,the peaks at 455 and 487 nm diminished and a new peak appearedat 390 nm. (Received February 12, 1983; Accepted June 20, 1983)     

Isolation of Subunits of Coupling Factor 1 from Maize and Spinach Chloroplasts and Properties of Combinations of Subunits with ATPase Activity

Subunits (, ß, ) and mixtures of subunits ( ß, , ß , ß ) were isolated without denaturationfrom a chloroform extract of chloroplast coupling factor 1 (CF₁)from maize (Zea mays var. Ushiku 5-4) and from spinach by fastprotein liquid chromatography (FPLC), on an anion-exchange columnof Mono-Q in the presence of n-octylglucoside (OG) and on achromatofocusing column of Mono-P. The ß -subunitcomplex (CF¹ ß ) was the minimum unit required forATPase activity, as was confirmed by the reconstituted complexof ß and subunits. An subunit isolated from maizeinhibited the ATPase activity of CF₁ ß from bothmaize and spinach. CF₁ ß was found to contain anOG-dependent Mg²⁺-ATPase. The ATPase activity of CF₁ ß required divalent cations, such as Mg²⁺ or Mn²⁺, for its expressionin the presence of OG; its optimum pH was 8.0 and it was markedlyinhibited by NaN₃. The enzyme hydrolyzed ATP in prefernece toGTP but not CTP, UTP, ADP, AMP or pNPP. Lineweaver-Burk plotsof its activity were curvilinear in the range of 0.6–0.7mM ATP.Mg²⁺. ¹Present address: Department of Biology, School of Education,Waseda University, Shinjuku-ku, Tokyo, 160 Japan. (Received February 15, 1989; Accepted April 20, 1989)     

Purification and properties of soluble cytochrome b-560 from spinach leaves

A b-type cytochrome having an -band at 560 nm was isolated fromspinach leaves (Spinacia oleracea). A method is described forpreparing this cytochrome, cytochrome b-560 (spinach), in apurified state. The cytochrome has, in its reduced state, absorption bands at560 nm (), 530 nm (ß) and 427 nm (); and in the oxidizedstate at 562 nm (), 529 nm (ß) and 417 nm (). Thepyridine ferro-haemochrome prepared from cytochrome b-560 hadan -band at 556.5 nm, indicating the protohaem-nature of theprosthetic group. The cytochrome has an oxidation-reduction potential (E'⁰) of+0.13V at pH 7.0, as measured using the ferri-ferro oxalate system. The cytochrome is rapidly reduced on illumination with red orfar-red light in the presence of spinach chloroplasts and isoxidized at a slower rate in the dark. This photoreduction isinhibited by 1x10^–6 M 3-(3,4-dichlorophenyl)-1,1-dimethylurea(DCMU). The molecular weight of the cytochrome is 30,000 asestimated by the dextran gel filtration method. (Received December 3, 1971; )     

r{beta}Carotene Hydroxylase Gene from the Cyanobacterium Synechocystis sp. PCC6803

The ORF sll1468 of Synechocystis sp. PCC6803 was identifiedas a gene for rß-carotene hydroxylase by functionalcomplementation in a rß-carotene-producing Escherichiacoll. The gene product of ORF sll11468 added hydroxyl groupsto the rß-ionone rings of rß-carotene (rß,rß-carotene)to form zeaxanthin (rß,rß-carotene-3,3'-diol).This newly identified rß-carotene hydroxylase doesnot show overall amino acid sequence similarity to the knownrß-carotene hydroxylases. However, it showed significantsequence similarity to rß-carotene ketolases of marinebacteria and a green alga. (Received November 29, 1997; Accepted March 6, 1998)     

Effect of CPTA and of Nicotine on Chlorophyll and Carotenoid Contents of an Ornamental Gourd

The three coloured parts: green, yellow-green and yellow offruits of an ornamental gourd contained carotenoids and chlorophyllsin decreasing amounts, while the orange parts had only smallamounts of carotenoids. There were 14 individual carotenoids(typically leaf carotenoids) in the green parts with three cis-xanthophylls(normally found in maturing fruits) which were in smaller amountsin the other coloured parts and disappeared completely in theorange parts. When the fruits were stored for 15 days, both carotenoids andchlorophylls (where present) decreased. 2-(4-chlorophenylthio)-triethylaminehydrochloride (CPTA)-treated fruits unusually contained eitherslightly less or slightly more carotenoids than controls, whilechlorophylls were always less. Lycopene and -carotene, absentin controls, accumulated in the treated fruits. These resultssupport the suggestion that CPTA may work as a cyclization reactioninhibitor. Nicotine however, inhibited total chlorophylls but increasedtotal carotenoids without accumulating either -carotene or lycopene.The results cannot be explained on the basis that only the non-ionicform of nicotine is effective as a carotenoid cyclization inhibitor. Ornamental gourd, Cucurbita, carotenoids, lycopene, xanthophylls, chlorophylls, 2-(4-chlorophenylthio)-triethylamine hydrochloride     

Carotenoid glucosides and menaquinones from the gliding bacterium Herpetosiphon giganteus Hp a2

The gliding bacterium Herpetosiphon giganteus Hp a2 was shown to contain -carotene, one monoglucosyloxy and two new diglucosyloxy carotenoids with a x-O-acyl-diglucosyloxy carotenoid as main component. Flexirubin-like pigments could not be detected in this organism.As in the Myxobacterales and Cytophagales (Kleinig et al., 1974) menaquinones (MK-6 and MK-7) were found to be the only isoprenoid quinones present in Herpetosiphon.The chemosystematic implications of these findings are briefly discussed.