A transport system for phosphoenolpyruvate, 2-phosphoglycerate, and 3-phosphoglycerate in Salmonella typhimurium.

Salmonella typhimurium strain LT-2 was found to utilize phosphoenolpyruvate, 2-phosphoglycerate, and 3-phosphoglycerate as sole sources of carbon and energy for growth, but Escherichia coli strains did not. The following evidence suggests that this growth difference was due to the presence in Salmonella cells of an inducible phosphoglycerate permease distinct from previously studied transport systems: (a) The ability of cells to take up 3-phospho[14-C]glycerate was induced by growth in the presence of phosphoenolpyruvate, 2-phosphoglycerate, or 3-phosphoglycerate, but not glycerate, alpha-glycerophosphate, or other carbon sources tested. (b) Uptake of 3-phospho[14-C]glycerate was strongly inhibited by the three nonradioactive inducers of 3-phosphoglycerate uptake, but not by glycerate or alpha-glycerophosphate. (c) Mutants which lost the ability to utilize and take up 3-phosphoglycerate simultaneously lost the ability to utilize 2-phosphoglycerate and phosphoenolpyruvate, but not other compounds tested. (d) Mutant strains which constitutively synthesized the phosphoglycerate transport system could use both phosphoglycerates and phosphoenolpyruvate as sole sources of phosphate at low substrate concentrations. (e) A strain lacking alkaline and acid phosphatases could still grow with 3-phosphoglycerate as sole carbon source. Maximal rates of 3-phospho[14-C]glycerate uptake occurred at pH 6 in the presence of an exogenous energy source. The apparent Km for 3-phosphoglycerate uptake under these conditions was about 10-minus 4 M. The maximal uptake rate (but not the Km) was dependent on potassium ions. Although synthesis of the phosphoglycerate transport system appeared to be under adenosine 3:5-monophosphate control, glucose repressed induction only slightly. The genes controlling synthesis of the phosphoglycerate transport system (pgt genes) appeared to map at about 74 min on the Salmonella chromosome.     

Lipid metabolism during bacterial growth, sporulation, and germination: an obligate nutritional requirement in Bacillus thuringiensis for compounds that stimulate fatty acid synthesis.

The regulation of fatty acid biosynthesis by compounds that are required for growth of Bacillus thuringiensis was investigated using an vivo assay developed to measure fatty acid synthesis in germinating spores. A minimal glucose-ammonium-salts medium does not support growth even though previous radiorespirometric studies have shown B. thuringiensis to possess intact tricarboxylic acid and Embden-Meyerhof-Parnas pathways. Abundant growth does occur, however, when this medium is supplemented with either glutamate, aspartate, citrate, thiosulfate, cystine, or ethylenediaminetetraacetic acid. Cells held under nongrowing conditions incorporate acetate into fatty acids; fatty acid synthesis is stimulated by the compounds that permit growth. These alternate nutritional requirements are not manifestations of a vitamin or trace metal deficiency and do not reflect a chelation phenomenon. These results indicate a direct correlation between the capacity of these compounds to promote growth and to stimulate formation of fatty acids.     

Soluble and membrane-bound aspartate-binding activities in Salmonella typhimurium.

The specificities of the soluble and membrane aspartate-binding activities were compared with each other and with the specificity of aspartate chemotaxis and were found to be distinct. The soluble aspartate-binding protein was purified to homogeneity and had a molecular weight of 30,000. The dissociation constant was 10(-6) M for aspartate, and the protein bound glutamate, cysteic acid, and 2-amino-3-phosphonopropionate. Aspartate transport was inhibited by cysteic acid.     

Scanning electron microscopy of surface and internal features of developing perithecia of Neurospora crassa.

Stages in the development of perithecia of Neurospora crassa, designated by the time elapsed after crossing, were investigated with the scanning electron microscope, from protoperithecia through perithecia. The usual examination of external features of whole specimens with this instrument was augmented by a freeze-fracture technique which allowed the viewing of development internally as well. Rapid increases in perithecial size soon after crossing were followed by the appearance, in section, of a centrum, at first undifferentiated but subsequently developing ascogenous hyphae. The perithecial beak appeared as a compact mass easily distinguishable in whole specimens from the surrounding hyphae by means of texture as well as shape. Two ascospores were photographed during emergence from an ostiole, but ostioles were found more frequently closed than open.     

Response of three phytoplankton bioassay techniques in experimental ponds of known limiting nutrient

In a controlled enrichment study of eight experimental ponds, results from the batch bioassay, primary productivity incubation bioassay, and chemostat techniques for measuring limiting factors of phytoplankton algae were compared to the change in the natural system with nutrient addition. In the ponds, rapid and dramatic increase in both phytoplankton biomass and primary productivity upon the addition of nitrogen and phosphorus fertilizer offered conclusive evidence that these nutrients were limiting in the control ponds to which no nutrients were added. Both the batch bioassay and chemostat techniques clearly indicated nitrogen and possibly phosphorus as the limiting factors; however, the primary productivity incubation bioassay technique showed no increase in ¹⁴C uptake with addition of these nutrients. A species- and/or nutrient-specific time lag between nutrient uptake and increased carbon fixation is suggested to explain the failure of the technique to yield positive results within the 4-hour incubation period used.     

Effects of crp mutations on adenosine 3'',5''-monophosphate metabolism in Salmonella typhimurium.

Wild-type Salmonella typhimurium could not grow with exogenous cyclic adenosine 3',5'-monophosphate (AMP) as the sole source of phosphate, but mutants capable of cyclic AMP utilization could be isolated provided the parental strain contained a functional cyclic AMP phosphodiesterase.All cyclic AMP-utilizing mutants had the growth and fermentation properties of cyclic AMP receptor protein (crp) mutants, and some lacked cyclic AMP binding activity in vitro. The genetic defect in each such mutant was due to a single point mutation, which was co-transducible with cysG. crp mutants isolated by alternative procedures also exhibited the capacity to utilize cyclic AMP. crp mutants synthesized cyclic AMP at increased rates and contained enhanced cellular cyclic AMP levels relative to the parental strains, regardless of whether or not cyclic AMP phosphodiesterase was active. Moreover, adenylate cyclase activity in vivo was less sensitive to regulation by glucose, possibly because the enzyme II complexes of the phosphotransferase system, responsible for glucose transport and phosphorylation, could not be induced to maximal levels. This possibility was strengthened by the observation that enzyme II activity (measured both in vitro by sugar phosphorylation and in vivo by sugar transport and chemotaxis) was inducible in the parental strain but not in crp mutants. The results suggest that the cyclic AMP receptor protein regulates cyclic AMP metabolism as well as catabolic enzyme synthesis.     

Repression of Diaminopimelic Acid Decarboxylase in Escherichia coli: Gene Dosage Effects and Escape Synthesis

Gene dosage and escape synthesis experiments support the hypothesis that diaminopimelate decarboxylase repression by lysine involves a repressor molecule in a negative control system.     

Membrane-bound respiratory of Spirillum itersonii.

The membrane-bound respiratory system of the gram-negative bacterium Spirillum itersonii was investigated. It contains cytochromes b (558), c (550), and o (558) and beta-dihydro-nicotinamide adenine dinucleotide (NADH) and succinate oxidase activities under all growth conditions. It is also capable of producing D-lactate and alpha-glycerophosphate dehydrogenases when grown with lactate or glycerol as sole carbon source. Membrane-bound malate dehydrogenase was not detectable under any conditions, although there is high activity of soluble nicotinamide adenine dinucleotide: malate dehydrogenase. When grown with oxygen as the sole terminal electron acceptor, approximately 60% of the total b-type cytochrome is present as cytochrome o, whereas only 40% is present as cytochrome o in cells grown with nitrate in the presence of oxygen. Both NADH and succinate oxidase are inhibited by azide, cyanide, antimycin A, and 2-n-heptyl-4-hydroxyquinoline-N-oxidase at low concentrations. The ability of these inhibitors to completely inhibit oxidase activity at low concentrations and their effects upon the aerobic steady-state reduction levels of b- and c-type cytochromes as well as the aerobic steady-state reduction levels obtained with NADH, succinate, and ascorbate-dichlorophenolindophenol suggest that presence of an unbranched respiratory chain in S. itersonii with the order ubiquinone leads to b leads to c leads to c leads to oxygen.     

Mode of insertion of lipopolysaccharide into the outer membrane of escherichia coli.

A mutant of Escherichia coli that lacks uridine 5'-diphosphate galactose-4-epimerase makes lipopolysaccharide with less carbohydrate than the parent, unless galactose is present during growth. Carbohydrate is dense, and the outer membrane, which contains lipopolysaccharide, was found to be denser when isolated from cells grown with galactose then when galactose was omitted. Cells given galactose after growth in its absence rapidly formed dense regions within the outer membrane that disappeared when galactose was removed. These results indicate that lipopolysaccharide enters the outer membrane nonrandomly at a minimum of 10 to 22 discrete "insertion points." Isopycnic centrifugation provides a method for isolating these regions.     

INHIBITION OF CHLORELLA (CHLOROPHYCEAE) GROWTH BY FATTY ACIDS, USING THE PAPER DISC METHOD1,2

Using the paper disc-agar plate method, a number of fatty and related acids have been tested for tested activity for inhibiting the growth of Chlorella pyrenoidosa Chick. Of the saturated acids, a peak in growth inhibiting activity wax observed in the C₇–C₁₂ range, where inhibition wax observed when solutions down to 0.02 M were applied to the discs. Most of the unsaturated acids tested showed greater inhibition than did the corresponding saturated acids. Acrylic acid showed detectable inhibition at 0.001 M concentration.