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1.
Foss BJ Sliwka HR Partali V Naess SN Elgsaeter A Melø TB Naqvi KR O'Malley S Lockwood SF 《Chemistry and physics of lipids》2005,135(2):157-167
The water dispersibility of astaxanthin was greatly enhanced by converting it to a disodium disuccinate salt. This carotenoid salt behaved as a bolaamphiphile in water; dynamic light scattering (DLS) revealed the formation of stable aggregates with an average hydrodynamic radius close to 1 microm. Larger aggregates were observed in solutions of increased osmolarity. Absorption spectra demonstrated that the aggregates could withstand the addition of 20% acetonitrile before disintegrating to monomers. The physicochemical properties of this astaxanthin derivative in solution were comprehensively studied by measuring surface tension, critical aggregate concentration, surface concentration, molecule area, free energy of adsorption and micellation, adsorption-aggregate energy relationship, and equilibrium constants, and then compared with similar compounds reported previously in the literature. 相似文献
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Foss BJ Sliwka HR Partali V Naess SN Elgsaeter A Melø TB Naqvi KR 《Chemistry and physics of lipids》2005,134(2):85-96
The water dispersibility of a hydrophobic carotenoid has been greatly enhanced by using it as the acyl part in the synthesis of a highly unsaturated lysophospholipid. Dynamic light scattering has revealed the formation of stable aggregates with an average hydrodynamic radius of a few nanometers, and absorption spectra show that the aggregates can withstand the addition of ethanol or acetonitrile until the volume fraction of water falls below 70 and 62%, respectively. The properties of the carotenoid phospholipids have been characterized by determining surface tension, critical micelle concentration, surface concentration, molecular area, free energy of adsorption and micellation, adsorption-micellar energy relationship, and equilibrium constants. 相似文献
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Julian Willibald Stefanie Breukers Anjan Patel Christer L. Øpstad Stine Nalum Naess Vassilia Partali 《Chemistry and physics of lipids》2009,161(1):32-37
Stable cationic carotenoid aggregates — predominantly of the J-type — develop when the hydrochlorides of carotenoid aldoximes and ketoximes are exposed to water. The oxime hydrochlorides are obtained by simple syntheses from commercially available food color carotenoids. Bluish-purple, unstable transient compounds were observed during hydrochlorination performed at liquid nitrogen temperature. 相似文献
4.
A phytoene desaturase (PDS) gene was cloned and characterized from the unicellular green microalga Chlamydomonas reinhardtii. Functional complementation analysis revealed C. reinhardtii PDS (CrPDS) catalyzes the conversion of phytoene to the colored carotenoid ζ-carotene. A single amino acid substitution, L505F, enhanced its desaturation activity by 29%, as indicated by an in vitro enzymatic assay. In addition, CrPDS-L505F exhibited 27.7-fold higher resistance to the herbicide norflurazon. Glass bead-mediated delivery displayed a high transformation efficiency of C. reinhardtii with CrPDS-L505F, demonstrating clearly that the engineered endogenous CrPDS is a dominant selectable marker for C. reinhardtii and possibly for other green algae. Furthermore, the expression of PDS could enhance the intracellular carotenoid accumulation of transformants, opening up the possibility of engineering the carotenogenic pathway for improved carotenoid production in microalgae. 相似文献
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【背景】石油被称为“液体黄金”,人类的工业生产活动在利用其创造巨大社会价值的同时,也对自然环境造成了严重的污染。微生物修复技术是现阶段治理石油类污染有效的手段之一,具有经济、高效、无二次污染等优点。【目的】从受石油污染的土壤中分离高效降解长链烷烃正二十四烷的菌株,探究其降解特性及在微生物修复中的应用前景。【方法】通过形态学及16S rRNA基因测序进行菌株鉴定,采用气相色谱法检测菌株对正二十四烷的降解效果,并结合气相色谱-质谱(gas chromatography-mass spectrometer, GC-MS)分析降解中间产物以推测其潜在代谢途径。【结果】筛选到一株可高效降解正二十四烷的菌株C24MT1,经鉴定为不动杆菌属(Acinetobacter)。该菌株最适降解条件为30 °C、pH 9.0、盐度2 g/L,该条件下生长7 d对9 g/L正二十四烷的降解率高达86.63%;与此同时,菌株在强碱性环境(pH 11.0)中生长良好(OD600为0.39)并保持较高烷烃降解率(75.38%),对极端环境具备较强的耐受能力;对降解中间产物进行分析,推断菌株代谢长链烷烃正二十四烷的途径可能包括末端氧化及次末端氧化。【结论】不动杆菌C24MT1具有良好的环境适应能力及烷烃降解能力,在后续微生物菌剂开发和石油类污染土壤的环境修复领域具有巨大的应用前景。本研究可为盐碱地区高浓度石油类污染土壤的修复提供优良菌种,并进一步丰富石油烃类生物降解的菌种资源库。 相似文献
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Vaccination with the merozoite surface protein 3 (MSP3) of Plasmodium falciparum protects against infection in primates and is under development as a vaccine against malaria in humans. MSP3 is secreted and associates with the parasite membrane but lacks a predicted transmembrane domain or a glycosylphosphatidylinositol anchor. Its role in the invasion of red blood cells is unclear. To study MSP3, we produced recombinant full-length protein and found by size exclusion chromatography that the apparent size of MSP3 was much larger than predicted from its sequence. To investigate this, we used several biophysical techniques to characterize the full-length molecule and four smaller polypeptides. The MSP3 polypeptides contain a large amount of alpha-helix and random coil secondary structure as measured by circular dichroism spectroscopy. The full-length MSP3 forms highly elongated dimers and tetramers as revealed by chemical cross-linking and analytical ultracentrifugation. The dimer is formed through a leucine zipper-like domain located between residues 306 and 362 at the C terminus. Two dimers interact through their C termini to form a tetramer with an apparent association constant of 3 mum. Sedimentation velocity experiments determined that the MSP3 molecules are highly extended in solution (some with f/f(0) > 2). These data, in light of the recent discoveries of three other Plasmodium proteins containing very similar C-terminal sequences, suggest that the members of this newly identified family may adopt highly extended and oligomeric novel structures capable of interacting with a red blood cell at relatively long distances. 相似文献
7.
The title complex cation, [Sb(tbpc)(OH)(2)](+) (where tbpc denotes tetra(tert-butyl)phthalocyaninate, C(48)H(48)N(8)(2-)), has been prepared by oxidizing [Sb(tbpc)]I(3) with tert-butyl perbenzoate in CH(2)Cl(2), CHCl(3), o-dichlorobenzene and also without solvent in the range of 20-80 degrees C. This species has been isolated as I(3)(-) salt and characterized by elemental analysis, ESI-MS, FT-IR, optical absorption and emission, and magnetic circular dichroism spectroscopy. This compound is quite well soluble in common polar organic solvents (e.g., CH(2)Cl(2), acetonitrile, acetone) without detectable aggregation at least up to ca. 10(-4)M while much less soluble (e.g., benzene, chloronaphthalene) or insoluble (hexane) in non-polar solvents. Although this compound is insoluble in water, it makes hydrophilic colloids in acetone-water mixtures. The most intense absorption band (Q-band) in a specific solvent red-shifts with an increase in the refractive index of the solvent. However, considerable deviation of the Q-band positions in donor-solvents from linear correlation between the positions and Onsager's solvent polarity function suggests that there are significant specific chemical interactions between the axial hydroxyl groups and the surrounding donor molecules. The low fluorescence quantum yield (ca. 0.01) for [Sb(tbpc)(OH)(2)](+) suggests that the singlet excited state of this species is considerably quenched by the presence of antimony ion in the chromophore. 相似文献
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1. We used laboratory growth and feeding experiments to evaluate the role of ω3 highly unsaturated fatty acids (HUFA) in zooplankton nutrition. Polyunsaturated fatty acids (PUFA) comprised 40% of total fatty acids (FA) in the green alga Scenedesmus acutus but only 6% in the cyanobacterium Synechococcus elongatus.Scenedesmus contained modest amounts of the ω3 HUFA eicosapentaenoic acid (EPA, 20 : 5ω3) and docosahexaenoic acid (DHA, 22 : 6ω3), whereas Synechococcus contained only a trace of EPA and no DHA. 2. One-day-old Daphnia maintained high growth rates for 2–3 days on a diet of Synechococcus but exhibited reduced growth or even lost weight over the later part of the 6- or 7-day experiments. Daphnia magna grew better than D. galeata or D. pulicaria on a diet of pure Synechococcus. Daphnids fed the same concentration (0.5 mgC l–1) of green algae or a mixture of green algae and Synechococcus maintained high growth rates. 3. Supplementing Synechococcus with fish oil emulsions rich in ω3 HUFA markedly improved the growth and reproduction of all three Daphnia species. Supplementation with an emulsion of monounsaturated oleic acid (18 : 1ω9), however, did not affect the growth of D. galeata and caused a decrease in the growth of D. magna. 4. Short-term (7 min) and long-term (3 h) feeding trials with 14C-labelled Synechococcus were used to evaluate the effects of acclimation and mixed diets on carbon incorporation. D. galeata that had fed on unlabelled Synechococcus for 3 days exhibited no decline in clearance rate but a marked decline in carbon incorporation in comparison with animals acclimated with Scenedesmus or fed a mixture of Synechococcus and Scenedesmus. 5. Our results support the hypothesis that the poor nutritional quality of a cyanobacterium is at least partially due to a HUFA deficiency. Growth and feeding experiments both suggest that the utilization of Synechococcus is enhanced by HUFA from lipid reserves or a mixed diet. 相似文献
9.
As a hemoglobin (Hb)-based oxygen carrier (HBOC), Hb suffers from the disadvantages of short half-life, renal toxicity and vasoactivity. Because dextran is a macromolecule that can be easily derivatized with various chemical moieties, conjugation of Hb with dextran can effectively increase the size of Hb and overcome the disadvantages of Hb. Thus, a dextran-bovine Hb (bHb) conjugate (dex-bHb) was prepared by conjugation of bHb with periodate-oxidized dextran here. As an important functional amino acid residue of bHb, Cys-93(β) was reversibly protected by 4,4′-dithiodipyridine to avoid reaction with periodate-oxidized dextran. Dex-bHb showed significantly higher hydrodynamic volume and higher viscosity than bHb. Conjugation with dextran stabilized the R state of bHb and slightly altered the heme environment of bHb. Conjugation with dextran decreased the P50 of bHb, lowered the sensitivity to the allosteric effectors and slightly decreased the autoxidation rate of bHb. Thus, dex-bHb was expected to act as a potent HBOC with low oxidative toxicity. 相似文献
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Disodium disuccinate astaxanthin ('rac'-dAST; Cardax) is a water-dispersible C40 carotenoid derivative under development for oral and parenteral administration for cardioprotection of the at-risk ischemic cardiovascular patient. In experimental infarction models in animals (rats, rabbits, and dogs), significant myocardial salvage has been obtained, up to 100% at the appropriate dose in dogs. The documented mechanism of action in vitro includes direct scavenging of biologically produced superoxide anion; in vivo in rabbits, modulation of the complement activity of serum has also been shown. A direct correlation between administration of the test compound in animals and reductions of multiple, independent markers of oxidative stress in serum was recently obtained in a rat experimental infarction model. For the current study, it was hypothesized that oral Cardax administration would inhibit oxidative damage of multiple relevant biological targets in a representative, well-characterized murine peritoneal inflammation model. A previously developed mass spectrometry-based (LC/ESI/MS/MS) approach was used to interrogate multiple distinct pathways of oxidation in a black mouse (C57/BL6) model system. In vivo markers of oxidant stress from peritoneal lavage samples (supernatants) were evaluated in mice on day eight (8) after treatment with either Cardax or vehicle (lipophilic emulsion without drug) orally by gavage at 500 mg/kg once per day for seven (7) days at five (5) time points: (1) baseline prior to treatment (t=0); (2) 16 h following intraperitoneal (i.p.) injection with thioglycollate to elicit a neutrophilic infiltrate; (3) 4 h following i.p. injection of yeast cell wall (zymosan; t=16 h/4 h thioglycollate+zymosan); (4) 72 h following i.p. injection with thioglycollate to elicit monocyte/macrophage infiltration; and (5) 72 h/4 h thioglycollate+zymosan. A statistically significant sparing effect on the arachidonic acid (AA) and linoleic acid (LA) substrates was observed at time points two and five. When normalized to the concentration of the oxidative substrates, statistically significant reductions of 8-isoprostane-F(2alpha) (8-iso-F(2alpha)) at time point three (maximal neutrophil recruitment/activation), and 5-HETE, 5-oxo-EET, 11-HETE, 9-HODE, and PGF(2alpha) at time point five (maximal monocyte/macrophage recruitment/activation) were observed. Subsequently, the direct interaction of the optically inactive stereoisomer of Cardax (meso-dAST) with human 5-lipoxygenase (5-LOX) was evaluated in vitro with circular dichroism (CD) and electronic absorption (UV/Vis) spectroscopy, and subsequent molecular docking calculations were made using mammalian 15-LOX as a surrogate (for which XRC data has been reported). The results suggested that the meso-compound was capable of interaction with, and binding to, the solvent-exposed surface of the enzyme. These preliminary studies provide the foundation for more detailed evaluation of the therapeutic effects of this compound on the 5-LOX enzyme, important in chronic diseases such as atherosclerosis, asthma, and prostate cancer in humans. 相似文献
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5-Ethylphenazine-poly(ethylene glycol)-NAD+ conjugate (EP+-PEG-NAD+) was prepared by linking 1-(3-carboxypropyloxy)-5-ethylphenazine (I) to poly(ethylene glycol)-bound NAD+ (PEG-NAD+) and its kinetic properties were studied. As a reference compound, poly(ethylene glycol)-bound 5-ethylphenazine derivative (III) was also prepared and the effects of poly(ethylene glycol) on the reaction rate of the 5-ethylphenazine moiety with NADH was investigated. The second-order rate constant, k1, of the reaction of III with NADH is 2.78 mM-1 s-1 and is about 1.7 times that of 1-(3-ethoxycarbonylpropyloxy)-5-ethylphenazine (II) with NADH. A similar effect of the attached poly(ethylene glycol) was observed for the reaction of PEG-NADH with I or II. The second-order rate constants, k2 and k3, of the reactions of the reduced form of III with oxygen and with 3-(4',5'-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium ion, respectively, were k2 = 1.22 mM-1 s-1 and k3 = 32 mM-1 s-1; the k2 value is not changed but the k3 value is decreased by the attachment of the polymer. EP+-PEG-NAD+ works as a unique catalyst having an intramolecular reaction step within its turnover cycle in a coupled multi-step reaction system containing malate dehydrogenase, malate, EP+-PEG-NAD+, a tetrazolium salt and oxygen. The first-order rate constant, k4, of the intramolecular reaction was 1.1 s-1. The effects of the covalent linking of the 5-ethylphenazine and the NAD+ moieties were estimated by comparing the value of k4 with that of k1 for the reaction of III with NADH; the effective concentration of the NADH moiety for the 5-ethylphenazine moiety on the same EP+-PEG-NADH molecule (or vice versa) was calculated to be 0.40 mM from the ratio of k4/k1. The values of the rate constants in the coupled multi-step reaction system enable us to understand the dynamic features of the system and the characteristics of EP+-PEG-NAD+ as a catalyst are discussed. 相似文献
15.
Isolation, properties and amino acid sequence of a long-chain neurotoxin, Acanthophis antarcticus b, from the venom of an Australian snake (the common death adder, Acanthophis antarcticus). 总被引:1,自引:1,他引:0
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The venom of an Australian elapid snake, the common death adder (Acanthophis antarcticus), was chromatographed on a CM-cellulose CM52 column. One of the neurotoxic components, Acanthophis antarcticus b (toxin Aa b) was isolated in about 9.4% (A280) yield. The complete amino acid sequence of toxin Aa b was elucidated. Toxin Aa b is composed of 73 amino acid residues, with ten half-cystine residues, and has a formula weight of 8135. Toxin Aa b has no histidine or methionine residue in its sequence. The amino acid sequence of toxin Aa b is homologous with those of other neurotoxins with known sequences, although it is novel in having a valine residue at its N-terminus and an arginine residue at position-23, where a lysine residue is found in almost all the so-far-known neurotoxins. Irrespective of the latter replacement, the toxin Aa b is fully active, with an LD50 value (in mice) of 0.13 microgram/g body weight on intramuscular injection. 相似文献
16.
Quan C Alcala E Petkovska I Matthews D Canova-Davis E Taticek R Ma S 《Analytical biochemistry》2008,373(2):179-191
The glycated form of a basic recombinant humanized monoclonal antibody (rhuMAb) was separated and quantitated by boronate affinity chromatography using optimized shielding reagents. Characterization on the isolated glycated material by peptide mapping analysis, using liquid chromatography-mass spectrometry (LC-MS) and tandem mass spectrometry (MS/MS) sequencing techniques, identified eight reactive lysine primary amine sites. The glycation reaction extent was similar among the various reactive sites, ranging from approximately 1 to 12%, and a single histidine residue separated the most and least reactive sites. Boronate chromatography run in a linear gradient mode separated monoglycated rhuMAb from higher order glycated species and indicated that the majority ( approximately 90%) of glycated rhuMAb is monoglycated. Low-level glycation on a heavy chain lysine located within a complementarity-determining region (CDR) did not significantly affect binding activity in potency measurements. The glycated forms also behaved as slightly more acidic than the nonglycated antibody in charge-based separation techniques, observable by capillary isoelectric focusing (cIEF) and ion exchange chromatography (IEC). The boronate column has significantly increased retention of aggregated rhuMAb material under separation conditions optimized for the monomer form. Recombinant protein glycation initially occurred during production in mammalian cell culture, where feed sugar and protein concentrations contribute to the total overall glycation on this antibody product. 相似文献
17.
Paulovicová E Bystrický S Machová E Bujdáková H 《FEMS immunology and medical microbiology》2008,53(3):421-428
Candida albicans expresses a CR3-related protein (CR3-RP) antigenically, structurally and functionally related to human adhesion glycoprotein, also known as Mac-l, the iC3b receptor, or complement receptor type 3. The purpose of the present study was to analyze the immunogenic properties of a novel CR3-RP glycoconjugate in a rabbit model. Cell-mediated responses revealed immunoenhancement triggered by CR3-RP glycoconjugate with respect to expression of IL-2 receptor subunit CD25 on B-lymphocytes and inductive increase of the CD4(+)/CD8(+) ratio compared with unconjugated cell wall mannan (P<0.001). Active immunization with the CR3-RP glycoconjugate resulted in an effective IgM-IgG isotypic switch even after the second booster. Altogether, it could be assumed that the novel peptide glycoconjugate is a prospective antigenic candidate for further Candida vaccine design. 相似文献
18.
Hog thyroid peroxidase: physical, chemical, and catalytic properties of the highly purified enzyme. 总被引:4,自引:0,他引:4
A B Rawitch A Taurog S B Chernoff M L Dorris 《Archives of biochemistry and biophysics》1979,194(1):244-257
Studies are reported on the purity and on the physical, chemical, and catalytic properties of a highly purified, stable, thyroid peroxidase (TPO). The enzyme was solubilized by treatment with deoxycholate and trypsin, and it was purified by a series of column treatments, including ion-exchange chromatography on DEAE-cellulose, gel filtration through Bio-Gel P-100, and hydroxylapatite chromatography. The final product, designated TPO VII, had a value for A410/A280 of 0.54, and its specific activity based on the guaiacol assay (794 μmol of guaiacol oxidized/min/mg) was considerably greater than that of any previously described TPO. Specific activity values based on other peroxidase-catalyzed reactions were also higher for TPO VII than for previous TPO preparations. Purity estimates for TPO VII, based on polyacrylamide disc gel electrophoresis and on isoelectric focusing in polyacrylamide gels, ranged from 80 to 95%. The molecular weight, determined by sedimentation equilibrium, was 93,000. Results of sodium dodecyl sulfate-gel electrophoresis also indicated a molecular weight of approximately 90,000. Sodium dodecyl sulfate-gel electrophoresis under reducing conditions indicated that TPO VII is composed of two peptide chains of unequal size, with the larger about 2.5-fold the size of the smaller. Carbohydrate analysis revealed that TPO is a glycoprotein containing about 10% by weight of carbohydrate. The predominant sugars were mannose and N-acetyl glucosamine. A significant amount of glucose was also found, along with small amounts of galactose, fucose, and xylose. The amino acid composition of TPO VII showed a high proline content, a predominance of arginine over lysine, and a ratio of [Asp] plus [Glu] to [Lys] plus [Arg] of over 2. Isoelectric focusing in polyacrylamide gels indicated an isoelectric pH of 5.75. In agreement with observations made on earlier preparations of TPO, heme spectral data showed significant differences between the pyridine hemochromogens of TPO VII and horseradish peroxidase, suggesting that the heme in TPO is not ferriprotoporphyrin IX. Circular dichroism measurements indicated that approximately 40% of TPO VII involves α helix or β structure. 相似文献
19.
4-methyleneglutamine amidohydrolase from peanut leaves : preparation, catalytic properties, and immunological responses of a highly purified form of the enzyme 总被引:1,自引:0,他引:1
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4-Methyleneglutamine amidohydrolase has been extracted and purified over 1000-fold from 14-day-old peanut (Arachis hypogaea) leaves by modification of methods described previously. The purified enzyme shows two bands of activity and three to four bands of protein after electrophoresis on nondenaturing gels. Each of the active bands is readily eluted from gel slices and migrates to its original position on subsequent electrophoresis. Although they are electrophoretically distinct, the two forms of the enzyme are immunologically identical by Ouchterlony double-diffusion techniques and have similar catalytic properties. Activity toward glutamine that has a threefold lower Vmax and a four-fold higher Km value copurifies with MeGln aminohydrolase activity. 4-Methyleneglutamine and 4-methyleneglutamic acid inhibit the hydrolysis of glutamine while glutamine inhibits 4-methyleneglutamine hydrolysis, further indicating the identity of the activity toward both substrates. Amidohydrolase activity is stimulated up to threefold by preincubation with either ionic or non-ionic detergents (0.1%) and also by added proteins (0.5% bovine serum albumin or whole rabbit serum); it is inhibited 50% by 1 millimolar borate or the glutamine analog, albizziin (10 millimolar). Rabbit antiserum to the purified peanut enzyme cross-reacts with one or more proteins in extracts of some plants but not others; in no instance, however, was 4-methyleneglutamine amidohydrolase activity detected in other species. Overall, the results support the hypothesis that 4-methyleneglutamine supplies N, via its hydrolysis by the amidohydrolase, to the growing shoots of peanut plants, whereas glutamine hydrolysis is prevented by the prepon-derance of the preferred substrate. Some results also suggest that this amidohydrolase activity may be regulated by metabolites and/or by association with other cellular components. 相似文献
20.
Scleractinian corals have adapted to live in habitats were the level of ultraviolet radiation (UVR, 280–400 nm) is extremely high. The putative photoprotective molecules called mycosporine-like amino acids (MAAs) contained in the corals' tissues absorb UVR and release it harmlessly as heat. MAA concentration in corals is quite plastic and correlates well with UVR dose, but other ecological factors such as water motion may influence MAA production as well. In this study, the effects of ambient UVR and water motion on MAA concentration and several physiological parameters of the reef coral Porites compressa Dana were investigated in a two by two factorial transplantation experiment. Replicate branches from nine morphologically distinct colonies were transplanted from the windward side of Coconut Island (Kaneohe Bay, HI) to a control area on the windward side (ambient water motion) and to an area on the leeward side (low water motion). The transplanted corals were placed under UV-opaque (UVO) or UV-transparent (UVT) filters fixed to the reef. Initially and at 3 and 6 weeks, coral branches were weighed to determine calcification rate and tissues were extracted in methanol for photosynthetic pigment and MAA analysis via high performance liquid chromatography (HPLC). UVR was a significant factor determining MAA concentration. When UVR was screened from the corals' environment, total MAA concentration decreased by 33% over 6 weeks. However, UVR-exposed corals moved to low water motion also decreased MAA levels, while UVR-exposed corals moved to the control area retained initial levels. Photosynthetic pigments and calcification rate were also significantly reduced in corals moved to low water motion. There was no UVR effect on photosynthetic pigments or calcification rate. This study provides evidence that water motion is important for the maintenance of MAAs. However, there were interesting colony-specific patterns in MAA composition and response to the UVR treatment; some colonies had high total concentrations of MAAs in all treatments, while others displayed a pronounced UVR effect. Also, each genotype seemed to have its own signature MAA composition. These findings indicate a genetic (host, zooxanthellae or both) component to UVR resistance in this population of P. compressa. 相似文献