Physiological Studies of the Boron Requirement of the Diatom, Cylindrotheca fusiformis Reimann and Lewin

Cylindrotheca fusiformis, a marine pennate diatom, requiresboron for growth both in the light and in the dark. A syntheticsaline nutrient medium and plastic containers were used to demonstratethe boron requirement. A boron concentration of at least 0.5ppm supports the maximum growth-rate, with a generation timeof approximately 10 hours. Boron concentrations below 0.5 ppmresult in growth at a reduced rate, the effect being most apparentduring the first few days of growth of the culture. The growth-rateis also affected by the initial silicon: boron ratio in themedium, indicating some possible interaction between these twoelements.     

Effects of Boron Deficiency on Rubidium Uptake and Photosynthesis in the Diatom Cylindrotheca fusiformis

Culturing the diatom Cylindrotheca fusiformis under boron-deficient conditions leads to changes in ⁸⁶Rb uptake and photosynthesis prior to any effect on the rate of cell division. The influx rate of ⁸⁶Rb into boron-deficient cells was 79% of the control rate after 5 to 5.5 hours culture. Despite lowered ⁸⁶Rb influx, however, boron-deficient diatoms accumulated more ⁸⁶Rb than did control cells; this was due to the deficient cells' lower efflux rate. After 24 hours culture, boron-deficient cells had accumulated 30% more ⁸⁶Rb than had control cells, while releasing ⁸⁶Rb at only one-half the control rate. Increased photosynthetic rates were another effect of boron deficiency during this early stage of culture. Prior to 20 hours boron-deficient culture, diatoms had photosynthetic rates 37% greater than those of control cells. Corresponding to the increase in photosynthesis, boron-deficient diatoms had 12% more carbohydrate than control cells after 16 hours culture.     

皱肋文蛤氨基酸含量及脂肪酸组成分析

测定了皱肋文蛤(Meretrix lyrata)软体部分的氨基酸含量与脂肪酸组成.共检出17种氨基酸,总含量为软体部干重的52.26%;4种呈味氨基酸(天门冬氨酸、谷氨酸、甘氨酸和丙氨酸)的含量为22.07%,占氨基酸总量的42.23%;必需氨基酸(EAA)总含量为20.72%,其必需氨基酸的构成比例基本符合FAO/W...     

615小鼠血红蛋白α链的氨基酸组成及个别肽段的氨基酸序列

用CM-Cellulose-23柱层析分离纯化了615小鼠珠蛋白α链,测定其N端氨基酸残基为缬氨酸.615小鼠珠蛋白α链含有141个氨基酸残基,其中19个亮氨酸残基,10个组氨酸残基,9个缬氨酸残基,上述氨基酸残基的数目与文献中其亲本C₅₇BL不同.用胰蛋白酶水解615小鼠珠蛋白α链,发现有不溶性的‘核心’和可溶性的酶解片段.其中一个酶解肽段从N端数第8位氨基酸残基发生了突变,由亲本的缬氨酸变为亮氨酸.     

Photochemical Apparatus Organization in the Diatom Cylindrotheca fusiformis: Photosystem Stoichiometry and Excitation Distribution in Cells Grown under High and Low Irradiance

The photochemical apparatus organization in the thylakoid membraneof the diatom Cylindrotheca fusiformis was investigated in cellsgrown under high and low irradiance. High light (HL, 200µE.m^–2.s^–1)grown cells displayed a relatively low fucoxanthin to chlorophyll(Chl) ratio, a low photosystem (PS) stoichiometry (PSII/PS I=1.3/1.0)and a smaller photosynthetic unit size in both PS I and PS II.Low light (LL, 30µE.m^–2.s^–1) grown cells displayeda 30% elevated fucoxanthin content, elevated PS II/PS I=3.9/1.0and larger photosynthetic unit size for PS II (a change of about100%) and for PS I (by about 30%). In agreement, SDS polyacrylamidegel electrophoresis of thylakoid membrane polypeptides showedgreater abundance of PS I, RuBP carboxylase and ATP synthasepolypeptides in HL cells. In contrast, LL grown cells exhibitedgreater abundance of light-harvesting complex polypeptides.Assuming an efficiency of red (670 nm) light utilization of1.0, the measured efficiency of blue (481 nm) light utilizationwas 0.64 (HL cells) and 0.72 (LL cells). The lower efficiencyof blue versus red light utilization is attributed to the quenchingof absorbed energy by non-fucoxanthin carotenoids. Differencesin the efficiency of blue light utilization between HL and LLgrown cells are attributed to the variable content of fucoxanthin.The results support the hypothesis of a variable Chl a-Chl c-fucoxanthinlight-harvesting antenna associated with PS II and PS I in Cylindrotheca. (Received February 10, 1988; Accepted April 6, 1988)     

Assessment of Fatty Acid and Amino Acid Composition of Macroalgae from the Persian Gulf to Characterize their Suitability for Nutritional Supplements

Biology Bulletin - Macroalgae, commonly known as seaweed, can be found worldwide. Seaweeds offer a novel dietary ingredient because of their excellent biochemical composition and health protective...     

Role of Silicon in Diatom Metabolism : Cyclic Nucleotide Levels, Nucleotide Cyclase, and Phosphodiesterase Activities during Synchronized Growth of Cylindrotheca fusiformis

Adenylate cyclase, guanylate cyclase, and the cyclic nucleotide phosphodiesterases of Cylindrotheca fusiformis were characterized in crude and partially purified preparations. Both cyclases were membrane-bound and required Mn²⁺ for activity, though Mg²⁺ gave 50% activity with adenylate cyclase. Properties of adenylate cyclase were similar to those of higher eukaryotic cyclases in some respects, and in other respects were like lower eukaryotic cyclases. Guanylate cyclase was typical of other lower eukaryotic enzymes.

Two phosphodiesterase activities were found, one selective for cyclic AMP, the other for cyclic GMP. The 5′-nucleoside monophosphate was the major product of both activities and each of the enzymes had distinctive divalent cation requirements, pH optima, and kinetic parameters. Both phosphodiesterases were similar to those of other lower eukaryotes with one notable difference: the cyclic AMP enzyme was inhibited by calcium.

Changes in the cyclic nucleotide levels were quantitated in light-dark and silicon-starvation synchronized cultures using a more sensitive radioimmunoassay than used in a previously published study (Borowitzka and Volcani 1977 Arch Microbiol 112: 147-152). Contrary to the previous report, the cyclic GMP level did not change significantly in either synchrony. The cyclic AMP level increased dramatically very early in the period of DNA replication with the peak cyclic AMP accumulation substantially preceding that of DNA synthesis in both synchronies. There was no significant change in the activity of either cyclase or either phosphodiesterase during either synchrony. Thus, the mechanism for the rise in cAMP level remains unclear.

     

Short-Term and Long-Term Responses of Crassulacean Acid Metabolism Plants to Elevated CO(2)

For the leaf succulent Agave deserti and the stem succulent Ferocactus acanthodes, increasing the ambient CO₂ level from 350 microliters per liter to 650 microliters per liter immediately increased daytime net CO₂ uptake about 30% while leaving nighttime net CO₂ uptake of these Crassulacean acid metabolism (CAM) plants approximately unchanged. A similar enhancement of about 30% was found in dry weight gain over 1 year when the plants were grown at 650 microliters CO₂ per liter compared with 350 microliters per liter. Based on these results plus those at 500 microliters per liter, net CO₂ uptake over 24-hour periods and dry weight productivity of these two CAM succulents is predicted to increase an average of about 1% for each 10 microliters per liter rise in ambient CO₂ level up to 650 microliters per liter.     

SuccSite: Incorporating Amino Acid Composition and Informative k-spaced Amino Acid Pairs to Identify Protein Succinylation Sites

Protein succinylation is a biochemical reaction in which a succinyl group (-CO-CH2-CH2-CO-) is attached to the lysine residue of a protein molecule. Lysine succinylation plays important regulatory roles in living cells. However, studies in this field are limited by the difficulty in experimentally identifying the substrate site specificity of lysine succinylation. To facilitate this process, several tools have been proposed for the computational identification of succinylated lysine sites. In this study, we developed an approach to investigate the substrate specificity of lysine succinylated sites based on amino acid composition. Using experimentally verified lysine succinylated sites collected from public resources, the significant differences in position-specific amino acid composition between succinylated and non-succinylated sites were represented using the Two Sample Logo program. These findings enabled the adoption of an effective machine learning method, support vector machine, to train a predictive model with not only the amino acid composition, but also the composition of k-spaced amino acid pairs. After the selection of the best model using a ten-fold cross-validation approach, the selected model significantly outperformed existing tools based on an independent dataset manually extracted from published research articles. Finally, the selected model was used to develop a web-based tool, SuccSite, to aid the study of protein succinylation. Two proteins were used as case studies on the website to demonstrate the effective prediction of succinylation sites. We will regularly update SuccSite by integrating more experimental datasets. SuccSite is freely accessible at http://csb.cse.yzu.edu.tw/SuccSite/.     

Fatty Acid Composition and the Oxidation of Low-Density Lipoproteins

The effect of the fatty acid composition of low-density lipoprotein (LDL) on copper-ion-catalyzed oxidation of isolated LDL was examined in 18 normolipidemic men. The decrease in LDL linoleic acid concentration (ΔL) during oxidation was found to be strongly correlated with initial LDL linoleic acid concentration (r = 0.976, n = 18, P < 0.001), whereas the production of thiobarbituric acid reacting substances (TBARS) was not. The concentration of oleic acid in LDL was then increased significantly (mean increase 20%, P < 0.05) in 8 male volunteers by a daily dietary supplement of rapeseed oil/muesli for 4 weeks. The mean delay before copper-ion-catalyzed production of conjugated dienes (the lag phase) was significantly (P < 0.001) greater in LDL isolated after the study period (67 min) than that of before the study period (40 mm). The rate of formation of conjugated dienes, ΔL and TBARS production during oxidation of LDL was not significantly altered by the rapeseed oil/muesli supplement. These results suggest that the linoleic acid content of LDL is a determinant of individual variability in LDL oxidation, and that a rapeseed oil/muesli dietary supplement reduces the susceptibility of LDL to oxidation.     

Relationship among Cellular Fatty Acid Composition,Amino Acid Uptake and Neomycin Formation in a Mutant Strain of Streptomyces fradiae

Neomycin-producing strains of Streptomyces fradiae, whose cellular fatty acid spectra are of iso 16: 0-type or normal 16: 0-type, had about two to six times larger amino acid and hexosamine pools than a neomycin-nonproducing strain which has the anteiso 15: 0-type cellular fatty acid spectrum. About 50 to 80 percent of the amount of extractable free amino acids were L-glutamic acid in either type of cells. The difference of pool size in these strains seems to be explained by the difference in ability for amino acid uptake. That is, the ability for L-glutamate uptake of anteiso 15:0-type cells was markedly reduced and accumulated glutamate was easily washed out by buffer. Glucose, magnesium ions and L-glutamate were essential for the formation of neomycin by washed cells and, therefore, even the mutant ST–5B of anteiso 15: 0-type could accumulate a large amount of glutamate and produce neomycin as far as it was grown in a medium containing a high concentration of glutamate. These results indicate that a large pool of glutamate is essential for the formation of neomycin and the fatty acid spectrum is a factor governing the capacity to accumulate L-glutamic acid.     

Lipid Fatty Acid Composition and Thermophilicity of Cyanobacteria

An analysis of lipid fatty acid composition in several unicellular and filamentous forms of mesophilic and thermophilic cyanobacteria was performed. At 47°C (the temperature of thermophilic cyanobacteria maintenance in the collection), the unicellular thermophilic Synechococcus strains were devoid of polyenoic acids as distinct from the mesophilic forms of this genus at the temperature of 20°C (the temperature of this cyanobacterial maintenance in the collection). In the thermophilic Synechococcus elongatusIPPAS B-267 strain, a decrease in temperature did not result in the occurrence of C18 polyenoic acids, but the quantitative relationship between the saturated and unsaturated fatty acids (S/U ratio) was decreased twofold. In contrast, the culturing of mesophilic strains at 25–32°C resulted in an increase in the S/U ratio due to an increase in the proportion of the 16:0 acid. In the Synechococcus IPPAS B-434 strain, this treatment resulted in a decrease in the relative content of monoenoic, mainly hexadecenoic, acids. The cyanobacterium Gloeobacter violaceus, which lacks thylakoids, and whose photosystems are formed in a cell membrane, contained polyenoic acids. The filamentous thermophilic cyanobacterium Phormidium laminosum, at the maintenance temperature of 47°C, did contain polyenoic acids, but their proportion was considerably lower than that in the filamentous mesophilic forms, such as Tolypothrix sp. and Spirulina platensis. A relative content of hexadecenoic acids in Ph. laminosum was higher than in the mesophilic forms. A possible role of hexadecenoic acids in the processes of adaptation of cyanobacteria to high temperatures is discussed. A relationship between the characteristics of fatty acid composition fixed by evolution and the changes caused by adaptation to a particular environment is considered.     

Stability of Lactic Acid Bacteria to Freezing as Related to Their Fatty Acid Composition

The viability of Streptococcus lactis and Lactobacillus sp. A-12 after freezing at -17°C for 48 h was better preserved when the cells were grown in medium supplemented with oleic acid or Tween 80 (polyoxyethylene sorbitan monooleate). A pronounced change in the cellular fatty acid composition was noted when the bacteria were grown in the presence of Tween 80. In S. lactis the ratio of unsaturated to saturated fatty acids increased from 1.18 to 2.55 and in Lactobacillus sp. A-12 it increased from 0.85 to 1.67 when Tween 80 was added to the growth medium. The antibiotic cerulenin markedly inhibited the growth of lactic acid bacteria in tomato juice (TJ) medium but had almost no effect on the growth of the bacteria in TJ medium containing Tween 80 (or oleic acid). The antibiotic inhibited markedly the incorporation of [1-¹⁴C]acetate but had no inhibitory effect on the incorporation of exogenous [1-¹⁴C]oleate (or [1-¹⁴C]palmitate) into the lipid fractions of lactic acid bacteria. Thus, the fatty acid composition of lactic acid bacteria, inhibited by the antibiotic cerulenin, can be modulated by exogenously added oleic acid (or Tween 80) without the concurrent endogenous fatty acid synthesis from acetate. The data obtained suggest that cerulenin inhibits neither cyclopropane fatty acid synthesis nor elongation of fatty acid acyl intermediates. The radioactivity of cells grown in the presence of [1-¹⁴C]oleate and cerulenin was associated mainly with cyclopropane Δ19:0, 20:0 + 20:1, and 21:0 acids. As a consequence, cerulenin caused a decrease in the ratio of unsaturated to saturated fatty acids in lactic acid bacteria as compared with cells grown in TJ medium plus Tween 80 but without cerulenin. Cerulenin caused a decrease in the viability of S. lactis and Lactobacillus sp. A-12 after freezing at -17°C for 48 h only when Tween 80 was present in the growth medium. We conclude that the sensitivity of lactic acid bacteria to damage from freezing can be correlated with specific alterations in the cellular fatty acids.     

Orientation of pigments and pigment-protein complexes in the diatom Cylindrotheca fusiformis. A linear-dichroism study

The orientation of pigments and pigment-protein complexes of the marine diatom Cylindrotheca fusiformis was studied by linear dichroism at 77 K. The technique of polyacrylamide gel squeezing was used to orient the diatom intact cells, their isolated thylakoid membranes and the three pigment-protein complexes: chlorophyll ac-fucoxanthin, chlorophyll ac and PS I complexes. The data indicate that specific orientation of various pigments exists at all structure levels. Tentative assignments of various features of the linear-dichroism spectra to the major photosynthetic pigments are presented. The orientation of the three pigment-protein complexes with respect to the thylakoid membrane plane and the major axis of the cell is also discussed.     

Lactate dehydrogenase from autotrophic and heterotrophic cells of the marine diatom Cylindrotheca fusiformis Reimann & Lewin.

Cultures of Cylindrotheca furisormis grown either autotrohpically or heterotrophically on lactate contained significant amounts of NAD-dependent L(+)-lactate dehydrogenase (EC 1.1.1.27). Polyacylamide gel electrophoresis of crude enzyme extracts revealed a single band which was indistinguishable between autotrohpic and heterotrohpic cells. The Km for lactate of partially purified preparations was lower under heterotrophic conditions. The specific activity in crude extracts was higher under autotrophic than heterotrophic conditions; it dropped precipitously when autotrophic cells were transferred to the dark, increasing again only in the presence of lactate. These and related observations suggest that this enzyme has at most only a minor role in the assimilation of lactate during heterotrophic growth on lactate.     

Relative Amino Acid Composition Signatures of Organisms and Environments

Background

Identifying organism-environment interactions at the molecular level is crucial to understanding how organisms adapt to and change the chemical and molecular landscape of their habitats. In this work we investigated whether relative amino acid compositions could be used as a molecular signature of an environment and whether such a signature could also be observed at the level of the cellular amino acid composition of the microorganisms that inhabit that environment.

Methodologies/Principal Findings

To address these questions we collected and analyzed environmental amino acid determinations from the literature, and estimated from complete genomic sequences the global relative amino acid abundances of organisms that are cognate to the different types of environment. Environmental relative amino acid abundances clustered into broad groups (ocean waters, host-associated environments, grass land environments, sandy soils and sediments, and forest soils), indicating the presence of amino acid signatures specific for each environment. These signatures correlate to those found in organisms. Nevertheless, relative amino acid abundance of organisms was more influenced by GC content than habitat or phylogeny.

Conclusions

Our results suggest that relative amino acid composition can be used as a signature of an environment. In addition, we observed that the relative amino acid composition of organisms is not highly determined by environment, reinforcing previous studies that find GC content to be the major factor correlating to amino acid composition in living organisms.     

Analysis of Fatty Acid Content and Composition in Microalgae

A method to determine the content and composition of total fatty acids present in microalgae is described. Fatty acids are a major constituent of microalgal biomass. These fatty acids can be present in different acyl-lipid classes. Especially the fatty acids present in triacylglycerol (TAG) are of commercial interest, because they can be used for production of transportation fuels, bulk chemicals, nutraceuticals (ω-3 fatty acids), and food commodities. To develop commercial applications, reliable analytical methods for quantification of fatty acid content and composition are needed. Microalgae are single cells surrounded by a rigid cell wall. A fatty acid analysis method should provide sufficient cell disruption to liberate all acyl lipids and the extraction procedure used should be able to extract all acyl lipid classes.With the method presented here all fatty acids present in microalgae can be accurately and reproducibly identified and quantified using small amounts of sample (5 mg) independent of their chain length, degree of unsaturation, or the lipid class they are part of.This method does not provide information about the relative abundance of different lipid classes, but can be extended to separate lipid classes from each other.The method is based on a sequence of mechanical cell disruption, solvent based lipid extraction, transesterification of fatty acids to fatty acid methyl esters (FAMEs), and quantification and identification of FAMEs using gas chromatography (GC-FID). A TAG internal standard (tripentadecanoin) is added prior to the analytical procedure to correct for losses during extraction and incomplete transesterification.