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1.
Strictly anaerobic bacteria were enriched and isolated from freshwater sediment sources in the presence and absence of sulfate with sorbic acid as sole source of carbon and energy. Strain WoSo1, a Gram-negative vibrioid sulfate-reducing bacterium which was assigned to the species Desulfoarculus (formerly Desulfovibrio) baarsii oxidized sorbic acid completely to CO2 with concomitant stoichiometric reduction of sulfate to sulfide. This strain also oxidized a wide variety of fatty acids and other organic compounds. A Gram-negative rod-shaped fermenting bacterium, strain AmSo1, fermented sorbic acid stoichiometrically to about equal amounts of acetate and butyrate. At concentrations higher than 10 mM, sorbic acid fermentation led to the production of pentanone-2 and isopentanone-2 (3-methyl-2-butanone) as byproducts. Strain AmSo1 fermented also crotonate and 3-hydroxybutyrate to acetate and butyrate, and hexoses to acetate, ethanol, hydrogen, and formate. The guanine-plus-cytosine content of the DNA was 41.8±1.0 mol%. Sorbic acid at concentrations higher than 5 mM inhibited growth of this strain while strain WoSo1 tolerated sorbic acid up to 10 mM concentration.  相似文献   
2.
NirD is part of the nitrite reductase complex NirBD that catalyses the reduction of nitrite to NH3 in nitrate assimilation and anaerobic respiration. The crystal structure analysis of NirD from Mycobacterium tuberculosis shows a double β‐sandwich fold. NirD is related in three‐dimensional structure and sequence to the Rieske proteins; however, it does not contain any Fe–S cluster or other cofactors that might be involved in electron transfer. A cysteine residue at the protein surface, conserved in NirD homologues lacking the iron–sulfur cluster might be important for the interaction with NirB and possibly stabilize one of the Fe–S centers in this subunit. Proteins 2012. © 2012 Wiley Periodicals, Inc.  相似文献   
3.
The diaminopimelic acid pathway of lysine biosynthesis has been suggested to provide attractive targets for the development of novel antibacterial drugs. Here we report the characterization of two enzymes from this pathway in the human pathogen Pseudomonas aeruginosa, utilizing structural biology, biochemistry and genetics. We show that tetrahydrodipicolinate N-succinyltransferase (DapD) from P. aeruginosa is specific for the L-stereoisomer of the amino substrate L-2-aminopimelate, and its D-enantiomer acts as a weak inhibitor. The crystal structures of this enzyme with L-2-aminopimelate and D-2-aminopimelate, respectively, reveal that both compounds bind at the same site of the enzyme. Comparison of the binding interactions of these ligands in the enzyme active site suggests misalignment of the amino group of D-2-aminopimelate for nucleophilic attack on the succinate moiety of the co-substrate succinyl-CoA as the structural basis of specificity and inhibition. P. aeruginosa mutants where the dapA gene had been deleted were viable and able to grow in a mouse lung infection model, suggesting that DapA is not an optimal target for drug development against this organism. Structure-based sequence alignments, based on the DapA crystal structure determined to 1.6 Å resolution revealed the presence of two homologues, PA0223 and PA4188, in P. aeruginosa that could substitute for DapA in the P. aeruginosa PAO1ΔdapA mutant. In vitro experiments using recombinant PA0223 protein could however not detect any DapA activity.  相似文献   
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The feasibility of genetically engineering soybean seed coats to divert metabolism towards the production of novel biochemicals was tested. The genes phbA, phbB, phbC from Ralstonia eutropha each under the control of the seed coat peroxidase promoter were introduced into soybean and the production of polyhydroxybutyrate (PHB) was assayed. The analysis of seed coats arising from 4 independent transformation events demonstrated that PHB was produced at a mean of 0.12% seed coat dried weight with individual values up to 0.36%. These values demonstrate that it is possible to metabolically engineer soybean seed coats.  相似文献   
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Internalization of activated signaling receptors by endocytosis is one way cells downregulate extracellular signals. Like many signaling receptors, the yeast alpha-factor pheromone receptor is downregulated by hyperphosphorylation, ubiquitination, and subsequent internalization and degradation in the lysosome-like vacuole. In a screen to detect proteins involved in ubiquitin-dependent receptor internalization, we identified the sphingoid base-regulated serine-threonine kinase Ypk1. Ypk1 is a homologue of the mammalian serum- and glucocorticoid-induced kinase, SGK, which can substitute for Ypk1 function in yeast. The kinase activity of Ypk1 is required for receptor endocytosis because mutations in two residues important for its catalytic activity cause a severe defect in alpha-factor internalization. Ypk1 is required for both receptor-mediated and fluid-phase endocytosis, and is not necessary for receptor phosphorylation or ubiquitination. Ypk1 itself is phosphorylated by Pkh kinases, homologues of mammalian PDK1. The threonine in Ypk1 that is phosphorylated by Pkh1 is required for efficient endocytosis, and pkh mutant cells are defective in alpha-factor internalization and fluid-phase endocytosis. These observations demonstrate that Ypk1 acts downstream of the Pkh kinases to control endocytosis by phosphorylating components of the endocytic machinery.  相似文献   
8.
Animal behaviour studies often require the diagnosis and statistical evaluation of patterns among individuals of a group. However, association values derived for all pairs of individuals are mathematically interrelated and, hence, difficult to evaluate. A general method of matrix comparison, called the Mantel test, is described which accounts for such interdependencies. In this test, two square difference matrices are compared to determine whether a statistical association exists between corresponding elements. For example, distances in one of the matrices might represent differences in songs for each pair of a group of birds, while distances in the other matrix could be the geographic distances between each of the same bird pairs; the test would assess whether or not birds close to one another have songs that are more similar than those pairs that are located further apart geographically. Distances in the matrices can be of a variety of sorts (depending on the application), including for instance geographic distances, morphological differences, or behavioural differences. The test has wide applicability in studies of animal behaviour, and we present three examples. First, dialects of splendid sunbirds (Nectarinia coccinigaster) were investigated to determine whether local or regional patterns of geographic variation in song were present, and whether birds with similar dialects were concentrated within the same habitat. Second, dominance hierarchies of white-throated sparrows (Zonotrichia albicollis) and dark-eyed juncos (Junco hyemalis oreganus) were analysed by constructing appropriate hypothesis matrices that were contrasted against a matrix summarizing behavioural interrelationships of flock members. In the third application, involving progressions of yellow baboons (Papio cynocephalus), an approach was developed to assess whether members of a given sex occur adjacent (or at least closer) to one another in progression order more often than expected by chance, and also whether individuals of a given sex tend to be found at or near the end of a progression more frequently than predicted. Other possible applications of the Mantel test are discussed, and a detailed computational example is included.  相似文献   
9.
1. Ionophore-induced osmotic swelling was used to study Cl- transport in isolated rat liver mitochondria. 2. Energy-dependent, neutral ionophore-induced swelling in Cl- salts at pH 7.2 required K+ and was preceded by a brief lag phase that was absent in chlorotributyltin-induced swelling. 3. Treatments that stimulated or inhibited mitochondrial K+/H+ exchange had qualitatively similar effects on both valinomycin-induced swelling and the associated lag phase. 4. The results suggest that valinomycin-induced Cl- permeability results from an interaction between the K+/H+ antiporter and neutral ionophore K+ complexes.  相似文献   
10.
Summary The anion transport across the red blood cell membrane is assumed to occur by ionic diffusion through dielectric pores which are formed by protein molecules spanning the red blood cell membrane. The access of anions to the dielectric pores is regulated by anion adsorption sites positioned at the entrances of the pores. The adsorption of small inorganic anions to the adsorption sites is facilitated by ionizing cationic groups setting up a surface potential at the respective membrane surfaces. Applying the transition state theory of rate processes, flux equations for the unidirectional flux were derived expressing the unidirectional flux as a function of the fractional occupancies of anion adsorption sites at both membrane surfaces.The basic properties of the transport model were investigated. The concentration-dependence and the pH-dependence of the unidirectional fluxes were shown to depend upon the surface charge density and upon the affinity of the transported anion species to the anion binding sites. The concentration-response and the pH-response of the unidirectional fluxes of different anion species may differ substantially even if the anion species are transported by the same anion transport system. The model predicts a characteristic behavior of the Lineweaver-Burk plot and of the Dixon plot.A comparison between computer simulated and experimentally determined flux curves was made. By choosing a suitable set of parameters, the anion transport model is capable of simulating the concentration-dependencies and the pH-dependencies of the unidirectional sulfate and chloride flux. It is sufficient to change one single constant in order to convert the sulfate transport system into a chloride transport system. Furthermore, the model is capable of predicting the inhibitory action of chloride on the sulfate transport system. No attempts were made to fit the experimental data to the model. The behavior of the model was qualitatively in accordance with the experimental results.  相似文献   
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