In vitro assessment of the cardioprotective, anti-diabetic and antioxidant potential of Palmaria palmata protein hydrolysates

The contribution of protein fraction and proteolytic enzyme preparation to the in vitro cardioprotective, anti-diabetic and antioxidant activity of Palmaria palmata protein hydrolysates was investigated. Aqueous, alkaline and combined aqueous and alkaline P. palmata protein fractions were hydrolysed with the food-grade proteolytic preparations, Alcalase 2.4 L, Flavourzyme 500 L and Corolase PP. The hydrolysates had angiotensin converting enzyme (ACE) and dipeptidyl peptidase (DPP) IV inhibitory activity with IC₅₀ values in the range 0.19–0.78 and 1.65–4.60 mg mL^?1, respectively. The oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) values ranged from 45.17 to 467.54 and from 1.06 to 21.59 μmol trolox equivalents/g, respectively. Furthermore, hydrolysates (1 mg mL^?1) were show to inhibit renin within the range 0–50 %. In general, Alcalase 2.4 L and Corolase PP hydrolysates of aqueous protein displayed the highest in vitro activity. The results indicate that protein fraction and enzyme preparation used have significant effects on in vitro biofunctional activity of the hydrolysates. This study demonstrates the potential of P. palmata protein hydrolysates as multifunctional functional food ingredients for the prevention/control of hypertension and type II diabetes.     

The reproductive response of the sea urchins Paracentrotus lividus (G.) and Psammechinus miliaris (L.) to a hyperproteinated macrophytic diet

The sea urchins Paracentrotus lividus and Psammechinus miliaris are submitted to the same environmental conditions in the Bay of Brest. The relationship between seasonal changes in food source quality and their gonad production was investigated in reproducing experimentally these conditions. In a first stage two macroalgae (Palmaria palmata and Laminaria digitata) were tested. P. miliaris showed a stronger preference for P. palmata and over a year-long experiment both urchins progressively preferred P. palmata. Seasonal variations in the chemical composition of P. palmaria were observed in the Bay of Brest: total carbohydrates were important and the relative maximum (about 50%) was reached between February and August; the lipid level was low and had a relative maximum of about 1% in June and August. Total protein in P. palmaria was high compared to other seaweeds: the maximum value (25%) was observed in June, which was probably due to the maintenance of nitrogen nutrient in the bay.In the second stage of the study, seasonal changes in biochemical components of ingestion and absorption of the two sea urchins were followed in the laboratory using a monospecific diet of P. palmaria. The patterns of total carbohydrates and lipid absorption were very similar for both sea urchin species. Carbohydrates were absorbed strongly and uniformly, year round. Lipid absorption mimicked the lipid nutrient pattern in the food source. Only changes in protein absorption varied slightly between the two urchin species. Protein absorption was maximal for both species in February and June, but the quantity of absorbed protein was significantly higher in P. miliaris than in P. lividus during February. This increase was concomitant with protein storage in the sea urchin gonads, which peaked in February for P. miliaris and in June for P. lividus. P. lividus had a higher gonad production efficiency, based on gonad yield. The comparison between in situ data and the experimental results suggests that an algal diet more nitrogenous than the in situ algal food source would benefit the herbivorous P. lividus, rather than the more omnivorous species P. miliaris. Although P. milaris has been described as a species with large gonad production potential, P. lividus appears to be a more suitable species for echiniculture conditions.     

Bioremediation potential of Palmaria palmata and Chondrus crispus (Basin Head): effect of nitrate and ammonium ratio as nitrogen source on nutrient removal

Palmaria palmata and Chondrus crispus were grown for 4 weeks in 1-L flasks at 10 °C to evaluate nutrient uptake and their potential application as nutrient biofilters in effluent from finfish culture. For greatest bioremediation benefit within an integrated system, we conclude that a seaweed biofilter using these species should be placed prior to bacterial biofiltration for exposure to greater proportions of ammonium than nitrate, though it is apparent that the productivity of both species is not influenced by the nitrogen source. Five combinations of ammonium– and nitrate–nitrogen were compared, each with a total N concentration of 300 μM (300:0, 270:30, 150:150, 30:270, 0:300). Molar nitrogen/phosphorus ratio was 10:1. The maximum growth rates were 8.9 and 6.0 % per day for P. palmata and C. crispus, respectively. For both species, the total nitrogen uptake was highest at 300 μM ammonium, 4.46 mgN gDW^?1 day^?1 for P. palmata and 3.40 mg?N? g?DW^?1?day^?1 for C. crispus. Over a 24-h period, 23–37 % of the available nitrate and 91–100 % of the available ammonium were taken up by P. palmata. In the same period, C. crispus took up 55–87 % of available nitrate and 89–100 % of ammonium. Tissue N in P. palmata was highest (4.1 %) at 270 and 300 μM ammonium, while the nitrogen source did not have a significant effect on the tissue N of C. crispus (mean of 4.6 %).     

Subcomponents C1q of the first component of guinea pig and mouse complement: Comparative study of their asparagine-linked sugar chains

Guinea pig and mouse C1q, subcomponents of the first component of complement, contained six asparagine-linked sugar chains on the C-terminal non-collagenous globular regions of each molecule. After N-acetylation and successive NaB³H₄-reduction of asparagine-linked sugar chains liberated by hydrazinolysis, their structure was analysed by sequential exoglycosidase digestion in combination with sugar composition analyses. The sugar chains of C1q molecules of both animals were very similar and composed of the biantennary complex type sugar chains with the following outer chains in various combinations: (± NeuNAcα → )Galß1 → GlcNAcß1 → and Galß1 → Galß1 → GlcNAcß1 →. These chain moieties were found to be linked to a common core structure of Manα1 → (Manα1 → )Manß1 → GlcNAcß1 → (Fucα1 → )GlcNAc.     

Carotenoids in red algae

The carotenoid composition of the following 8 species of red algae has been studied quantitatively and qualitatively: Bangia fuscopurpurea, Nemalion helminthoides, Bonnemaisonia hamifera (tetrasporophyte), Gigartina stellata, Rhodymenia palmata, Ceramium rubrum, Polysiphonia brodiaei, and Polysiphonia urceolata. Naturally occurring material of G. stellata, R. palmata, and P. brodiaei was investigated, while monoalgally cultured material was obtained from the remaining 5 species. α- and β-carotene, lutein, zeaxanthin, and small amounts of α- or β-cryptoxanthin were commonly present, but the two species P. brodiaei and P. urceolata were devoid of carotenoids containing α-ionone rings. Fucoxanthin was detected in naturally occurring material of B. fuscopurpurea, N. helminthoides, C. rubrum, and P. brodiaei, but could not be found in cultured material of the 3 first-mentioned species. The possible origin of the fucoxanthin is discussed as well as the biochemical and phylogenetic implications of the results obtained. As judged from the electronic spectrum of the total extracts, chlorophyll d was not present in any of the algae investigated. The total amounts of carotenoids and the ratio carotenoids: chlorophyll a were similar to those reported in previous work.     

Oligosaccharides on cathepsin D from porcine spleen

Cathepsin D from porcine spleen contained mannose (3.3%), glucosamine (1.4%), and mannose 6-phosphate (0.08%). Essentially all of the oligosaccharides of cathepsin D could be released by endo-β-N-acetylglucosaminidase H, pointing to oligomajmoside types of structures. Three neutral oligosaccharide fractions, containing 5, 6, and 7 mannose residues, respectively, were isolated by gel permeation chromatography on Bio-Gel P-2. Studies using exoglycosidase digestions and 500-MHz ¹H NMR spectroscopy revealed that their structures are [Manα1 → 2]_{0 or 1}Manα1 → 6[Manα1 → 3]Manα1 → 6[(Manα1 → 2)_{0 or 1}Manα1 → 3]Manβ1 → 4GlcNAcβ1 → 4 GlcNAc. These structures are identical to what have recently been proposed by Takahashi et al. for the major oligosaccharide units of cathepsin D from the same source (T. Takahashi P.G. Schimidt, and J. Tang (1983)J. Biol. Chem.258, 2819–2930), except for the occurrence of two isomeric oligosaccharides containing six mannoses. Only a part (3.4%) of the oligosaccharides were acidic, containing phosphates in monoester linkage. The phosphorylated oligosaccharides also consisted of oligomannoside-type chains which were analogous to, but more heterogeneous in size than the neutral oligosaccharides. Cathepsin D was bound to a mannose- and N-acetylglucosamine-specific lectin (mannan-binding protein) isolated from rabbit liver with the K_i value of 5.4 × 10^?6m.     

Stress tolerance and antioxidant enzymatic activities in the metabolisms of the reactive oxygen species in two intertidal red algae Grateloupia turuturu and Palmaria palmata

This investigation was designed to compare the differential stress tolerance in young thalli of two similar intertidal red seaweeds, Grateloupia turuturu Yamada and Palmaria palmata Kuntze, and to identify whether the invasive alga G. turuturu was more stress tolerant than P. palmata to cope with adverse environmental conditions. To do so, we measured the production of reactive oxygen caused by methyl viologen (MV) by assessing the oxidation of dichlorohydrofluorescein (DCFH) to dichlorofluorescein (DCF), the activities of reactive oxygen scavenging enzymes and the changes of the optimal fluorescence quantum yield (Fv/Fm) when the thalli of the two species were exposed to oxidative stresses caused by the addition of MV, H₂O₂, 3(3, 4-dichlorophenyl)-1,1-dimethyl urea (DCMU), heavy metal, changes of salinities, heat and freezing. Results demonstrated that the activities of superoxide dismutase (SOD) and peroxidase (POD) in G. turuturu were much higher than in P. palmata. Fv/Fm in G. turuturu was less sensitive than that in P. palmata to MV, H₂O₂, DCMU, heavy metal, salinity and heat stress, indicating that G. turuturu could be better acclimatized to changing environments and thus had a higher threshold for oxidative stress than P. palmata. G. turuturu was shown to be more sensitive to freezing treatment (− 20 °C), which explained why the appearance of G. turuturu was rarely reported in colder water environments.     

NONEQUILIBRIUM RATES OF PHOTOSYNTHESIS AND RESPIRATION UNDER DYNAMIC LIGHT SUPPLY1

To identify processes that might account for differences in growth rates of rhodophytes under constant and dynamic light supply, we examined nonequilibrium gas exchange by measuring time courses of photoinduction, loss of photoinduction, and respiration rates immediately after the light–dark transition. Using the rhodophyte species Palmaria palmata (Huds.) Lamour and Lomentaria articulata (Huds.) Lyngb., we compared the effects of growth-saturating constant photon flux density (PFD) (95 μmol photons · m^?2· s^?1) to those of a dynamic light supply modeled on canopy movements in the intertidal zone (25 μmol photons · m^?2· s^?1 background PFD plus light flecks of 350 μmol photons · m^?2· s^?1, 0.1 Hz). The time required for P. palmata and L. articulata to be fully photoinduced was not affected by the dynamics of light supply. L. articulata required only 6 min of illumination with either fluctuating or constant light to be completely induced compared to 20 min for P. palmata. The latter species also lost photoinduction more rapidly than did L. articulata in the dark. There was no significant decline in photoinduction state for either species at the background PFD. The time courses of respiration after illumination with constant and fluctuating light were significantly different for P. palmata but not for L. articulata when the total photon dose was equal. In general, gas exchange of P. palmata appeared to be particularly sensitive to the temporal distribution of light supply whereas that of L. articulata was sensitive to the amplitude of variations, being photoinhibited at high PFD. These results are discussed in terms of the different mechanisms of inorganic carbon acquisition in the two species.     

Evidence for Multiple Phototransduction Pathways in a Reef-Building Coral

Photosensitive behaviors and circadian rhythms are well documented in reef-building corals and their larvae, but the mechanisms responsible for photoreception have not been described in these organisms. Here we report the cloning, immunolocalization, and partial biochemical characterization of three opsins and four G proteins expressed in planulae of the Caribbean elkhorn coral, Acropora palmata. All three opsins (acropsins 1–3) possess conserved seven-pass transmembrane structure, and localize to distinct regions of coral planulae. Acropsin 1 was localized in the larval endoderm, while acropsin 2 was localized in solitary cells of the ectoderm. These rod-like cells displayed a remarkably polarized distribution, concentrated in the aboral end. We also cloned four A. palmata G protein alpha subunits. Three were homologs of vertebrate Gi, Go, and Gq. The fourth is presumably a novel G protein, which displays only 40% identity with the nearest known G protein, and we termed it Gc for “cnidarian”. We show that Gc and Gq can be activated by acropsins in a light-dependent manner in vitro. This indicates that at least acropsins 1 and 3 can form functional photoreceptors and potentially may play a role in color preference during settlement, vertical positioning and other light-guided behaviors observed in coral larvae.     

The Molecular mechanism of the action of <Emphasis Type="Italic">Helianthus tuberosus</Emphasis> L. polysaccharide

Evidence for immunoadjuvant activity of the Helianthus tuberosus L. polysaccharide was obtained in an antibody-producing cell model. Dectin-1 and TLR-6 were identified as major receptors required for biological activity of polysaccharide in a TNF-α stimulation model. The CR3 receptor was not implicated in polysaccharide recognition in the same model. Enzyme treatment of the H. tuberosus L. polysaccharide demonstrated the presence of ß-(1 → 4) and ß-(1 → 3) glycosidic bonds.     

Sequential changes in glycolipid expression during human B cell differentiation: enzymatic bases

We have previously reported that human B cell differentiation is accompanied by sequential changes in glycosphingolipid expression. Pre-B cells contain lacto-series type II chain-based glycolipids and GM3 ganglioside; mature/activated B cells do not synthesize lacto-series compounds but express GM3 and globo-series glycolipids (Gb3 and Gb4); terminally differentiated B cells, in addition to these compounds, also contain GM2 ganglioside. At the cell surface, Gb3, Gb4 and GM2 constitute stage-specific antigens. To elucidate the biosynthetic mechanism leading to these modifications we have compared activities of the glycosyltransferases involved in the core structure assembly and the first elongation steps of neo-lacto, ganglio- and globo-series glycolipids. These glycosyltransferase activities have been measured in B cell lines and normal B lymphocytes at various stages of differentiation. We first determined the optimal requirements of the four glycosyltransferases which wynthesize Lc3, GM3, Gb4 and GM2 glycolipids in B lymphocytes and then tested these enzymes and the Gb3 synthetase in the selected B cells. The following results were obtained: ß1 → 3N-Acetylglucosaminyltransferase (Lc3 synthetase) has a high activity in pro- and pre-B cells whereas it is undetectable in more differentiated cells; α2 → 3 sialyltransferase (GM3 synthetase) is activated from the pre-B cell stage to the terminally differentiated myeloma cells; α → 4 galactosyltransferase (Gb3 synthetase) is only detected in cells representing the late stages of B cell differentiation; ß1 → 3N-Acetylgalactosaminyltransferase (Gb4 synthetase) is only found in some lymphoblastoid cell lines, representative of activated B cells whereas the ß1 → 4 N-Acetylgalactosaminyltransferase (GM2 synthetase) has a high activity in these lymphoblastoid cell lines and in terminally differentiated myeloma cells. These results suggest that the sequential shifts in the three major glycosphingolipid series observed during B cell differentiation are mostly due to sequential activations of the corresponding glycosyltransferases.     

The effects of feeding frequency and symbiosis with zooxanthellae on the biochemical composition of Astrangia Danae Milne Edwards & Haime 1849

The coral Astrangia danae Milne Edwards & Haime 1849 occurs naturally with and without symbiotic algae and thus may have two sources of nourishment: (1) particles captured by the coral polyps, and (2) photosynthetic products translocated from their zooxanthellae. Symbiotic colonies may have both sources, and nonsymbiotic ones certainly have only the former. The relative importance of these two food sources was studied in the laboratory by examining the tissues of corals fed with frozen brine shrimp. Stock corals were fed once per week. Two to three weeks prior to each experiment, selected corals were placed on one of three feeding schedules: starved (S), fed once per week (1/wk), and fed three times per week (3/wk). The coral tissues were analyzed for protein, lipid, carbohydrate, and zooxanthellae content. Increased feeding frequency (1/wk → 3/wk) resulted in an increased tissue biomass and lipid to protein (L/P) ratio; starvation (1/wk → S) caused a decrease in these parameters. Symbiosis with zooxanthellae had an effect similar to increased feeding frequency in that the S and 1/wk symbiotic corals had a higher L/P ratio than comparable nonsymbiotic ones. There were no significant differences in L/P ratios between the 3/wk symbiotic and nonsymbiotic corals. Freshly collected colonies had a tissue composition most similar to the laboratory animals fed 3/wk. This result is consistent with the hypothesis that ingestion of solid food is the major nutritional source for A. danae in Narragansett Bay, Rhode Island, but our experiments suggest that the algae can have an important effect on tissue L/P ratios during times of food scarcity.     

Biological Activities of Fundamental,Carbohydrate Skeleton of Lipid A Containing Amide-linked 3-Hydroxytetradecanoic Acid

In the initial stage of hydrolysis, exo-ß-(l-→3)-d-glucanase from Basidiomycetes sp. QM 806 cleaved laminaran from Eisenia bicyclis with a pattern resembling an endo-hydrolase. Five kinds of intermediate gluco-oligosaccharides were separated by a combination of gel filtration and HPLC. They were shown to be 3²-O-ß-d-glucosyl-gentiobiose, 3²-O-ß-d-gentiobiosyl-gentiobiose, 3³O-ß-d-glucosyl-gentiotriose, 3⁴-ß-d-glucosyl-3²-O-gentiobiosyl-gentiobiose, and 3³-O-ß-d-gentio-biosylgentiotriose by enzymic hydrolysis and methylation analysis as well as by ¹³C NMR spectroscopy. As a result, such kinds of ß-(l → 3)-ß-(l→6)-linked oligosaccharides could be accounted for in the initial cleavage, and they were hydrolyzed ultimately to glucose, gentiobiose, and gentio-triose. It suggests that a single (1 → 3)-linkage on a block of (1 → 6)-links show some resistance to attack by this enzyme.     

Assessment of Acropora palmata in the Mesoamerican Reef System

The once-dominant shallow reef-building coral Acropora palmata has suffered drastic geographical declines in the wider Caribbean from a disease epidemic that began in the late 1970s. At present there is a lack of quantitative data to determine whether this species is recovering over large spatial scales. Here, we use quantitative surveys conducted in 107 shallow-water reef sites between 2010 and 2012 to investigate the current distribution and abundance of A. palmata along the Mesoamerican Reef System (MRS). Using historical data we also explored how the distribution and abundance of this species has changed in the northern portion of the MRS between 1985 and 2010–2012. A. palmata was recorded in only a fifth of the surveyed reef sites in 2010–2012. In the majority of these reef sites the presence of A. palmata was patchy and rare. Only one site (Limones reef), in the northernmost portion of the MRS, presented considerably high A. palmata cover (mean: 34.7%, SD: 24.5%). At this site, the size-frequency distribution of A. palmata colonies was skewed towards small colony sizes; 84% of the colonies were healthy, however disease prevalence increased with colony size. A comparison with historical data showed that in the northern portion of the MRS, in 1985, A. palmata occurred in 74% of the 31 surveyed sites and had a mean cover of 7.7% (SD = 9.0), whereas in 2010–2012 this species was recorded in 48% of the sites with a mean cover of 2.9% (SD = 7.5). A. palmata populations along the MRS are failing to recover the distribution and abundance they had prior to the 1980s. Investigating the biological (e.g., population genetics) and environmental conditions (e.g., sources of stress) of the few standing reefs with relatively high A. palmata cover is crucial for the development of informed restoration models for this species.     

High-resolution mass spectrometry confirms the presence of a hydroxyproline (Hyp) post-translational modification in the GGGGP linker of an Fc-fusion protein

Flexible and protease resistant (G4S)_n linkers are used extensively in protein engineering to connect various protein domains. Recently, several groups have observed xylose-based O-glycosylation at linker Ser residues that yield unwanted heterogeneity and may affect product quality. Because of this, an engineering effort was implemented to explore different linker sequence constructs. Here, we demonstrate the presence of an unexpected hydroxylation of a prolyl residue in the linker, made possible through the use of high-resolution mass spectrometry (HR-MS) and MS_n. The discovery started with the detection of a poorly resolved ～+17 Da mass addition at the reduced protein chain level of an Fc-fusion construct by liquid chromatography-MS. Upon further investigation at the peptide level using HR-MS, the mass increase was determined to be +15.99 Da and was localized to the linker peptide SLSLSPGGGGGPAR [210–223]. This peptide corresponds to the C-terminus of Fc [210–216], the G4P linker [217–221], and first 2 amino acids of a growth factor [222–223]. The linker peptide was first subjected to MS² with collision-induced dissociation (CID) activation. The fragmentation profile localized the modification to the GGGPA [218–222] portion of the peptide. Accurate mass measurement indicated that the modification is an addition of an oxygen and cannot be CH₄, thus eliminating several possibilities such as Pro→Leu. However, other possibilities cannot be ruled out. Higher-energy collision-induced dissociation (HCD)-MS² and MS³ using CID/CID were both unable to differentiate between Ala222→ Ser222 or Pro221→ Hyp221. Finally, MS³ using high-resolution CID/HCD confirmed the mass increase to be a Pro221→Hyp221 post-translational modification.     

Novel extraction method of antioxidant compounds from Sasa palmata (Bean) Nakai using steam explosion

Antioxidant compounds were extracted from various parts of Sasa palmata (Bean) Nakai, a bamboo plant whose leaves are commonly used to wrap foodstuffs such as Sushi in Japan. Highest concentrations of antioxidant compounds existed in the leaf part of S. palmata. Steam explosion treatment followed by hot water and methanol extractions was used for separating the antioxidant compounds from S. palmata leaf. The steam explosion treatment is the physical–chemical treatment which crushes a sample by sudden reduction of the pressure in reactor to atmospheric pressure after steaming the sample at high temperature and pressures. Sasa palmata leaf was hydrolyzed by steaming and crushed by the rapid decompression. The optimal condition of steam explosion for the effective extraction of antioxidant compounds from S. palmata was determined as a steam of temperature of 250 °C and a steaming time of 1 min. In these conditions 217.41 mg/(g-Sasa leaf) of phenolic compounds and 142.81 mg/(g-Sasa leaf) of radical scavenging activity, that was expressed as butylated hydroxyanisole (BHA), were obtained.     

Use of enzymatic cell wall degradation for improvement of protein extraction from Chondrus crispus,Gracilaria verrucosa and Palmaria palmata

The effect of polysaccharidases (κ-carrageenase, β-agarase, xylanase, cellulase) on the protein extraction from three rhodophytes has been studied. The kinetic parameters (apparent V _m, apparent K _m) and the optimum activity conditions (pH, temperature) of each enzyme were determined by using pure substrates. All the tested enzymes possess Michaelis Menten mechanism with estimated substrate saturating concentrations of 8 000 mg l⁻¹(carrageenan) for κ-carrageenase, 8 000 mg l⁻¹ (agar) for β-agarase, 5000 mg l⁻¹ (xylane) for β-xylanase and 6 000 mg l⁻¹ (carboxymethylcellulose) for cellulase. The optimum activity conditions are pH 6.5–6.8 at 45°C for carrageenase, pH 6–6.5 at 55°C for agarase, pH 5 at 55°C for xylanase and pH 3.8 at 50°C for cellulose. Different alga/enzymes couples (κ-carrageenase/Chondrus crispus, β-agarase/Gracilaria verrucosa, β-xylanase/Palmaria palmata) were tested under the optimum activity conditions. Alga/cellulase + specific enzyme (e.g. Chondrus crispus/carrageenase + cellulase) systems were also studied at the optimum activity conditions of a specific enzyme (e.g. carageenase). The use of the only cellulose was also tested on each alga. Except for Palmaria palmata, the highest protein yields were observed with the procedures using cellulase coupled with carrageenase or agarase for an incubation period limited to 2 h. The Chondrus crispus/carrageenase + cellulose and Gracilaria verrucosa/agarase + cellulase systems gave ten-fold and three-fold improvements, respectively, in protein extraction yield as compared to the enzyme-free blank procedure. The combined action of xylanase and cellulose on protein extraction from Palmaria palmata does not significantly improve protein yield. The best overall protein yield for P. palmata is for P. palmata/xylanase with a 14-h incubation time. This study shows the interest in the use of a polysaccharidase mixture for improving protein extractibility from certain rhodophytes. This biotechnology approach, adapted from procedures for protoplast production or enzymatic liquefaction of higher plants, could be tested as an alternative method to obtain proteins from seaweeds of nutritional interest.     

INORGANIC NITROGEN AND PHOSPHORUS UPTAKE KINETICS IN PALMARIA PALMATA (RHODOPHYTA)1

The N and P uptake responses were studied in a northern Spanish population of the edible red seaweed Palmaria palmata (Linnaeus) Kuntze. The fronds were incubated at different concentrations, and the nutrient depletion in the medium was measured at successive times to calculate uptake rates. Palmaria palmata uptake response was biphasic and nonsaturable for inorganic P. This would allow the species to exploit transient pulses of high P concentration in natural and fertilized conditions. Such a response is a common feature of algae avoiding nutrient deficiency. At average concentrations measured in the ocean, the response was nonsaturable for inorganic N sources, except for ammonium in autumn and winter when it is not the major N source. In contrast to the general rule of ammonium being taken at a higher rate than nitrate, we found similar affinity for both nutrients corresponding to the minor role of ammonium as N source for field populations over the year.     

Structure characterization of haemostatic diosgenin glycosides from paris polyphylia

From Paris polyphylla var. chinensis Hare (Liliaceae), four diosgenin glycosides with haemostatic effects were isolated. The structure of the major component was elucidated by chemical and spectroscopic methods as 3-{[α-L-rhamnopyranosyl(1_Rha → 2_Glu)]-[α-L-arabinofuranosyl(1_Ara → 4_Glu)]-β D-glucopyranosyl}-25(R)-spirost-5-en-3β-ol. This saponin was found to be identical to three previously reported compounds to which other structures were originally assigned, namely the major component from P. polyphylla Smith, the major cytotoxic component of yunnan paiyao, and polyphyllin D from P. polyphylla grown in the Himalaya region.     

Activity of fructanase in batch cultures of oral streptococci

Several strains of oral streptococci produced fructanase when grown in the absence of d-fructan in a complex medium supplemented with d-glucose. The major part of the activity was extracellular, and only 1–5% was associated with the cells. Release of fructanase began early in the exponential phase and the enzyme was stable in the stationary phase for several h if the pH did not fall below 6. Among the strains of Streptococcus mutans, serotypes a, d, and g released the highest amount of fructanase, and the low level of enzyme produced by strains of serotype c was increased when d-fructose replaced d-glucose as carbon source for growth. Fructanase of S. mutans readily hydrolysed (2 → 6)-β-d-fructans, but (2 → 1)-β-d-fructans and inulin were more resistant. Adsorption of fructanase to (2 → 6)-β-d-fructan, or inhibition with Tris buffer, provided effective means of eliminating fructanase activity from culture filtrates. This procedure should permit a more accurate determination of fructosyltransferase activity of S. mutans strains.