Hydrocarbon metabolism in a marine bacterium]

The marine bacterium L.16.1 (Alcaligenes sp.) grows preferentially on alkanes (C10 to C18) with a very high growth yield (98 per cent); optimal growth depends strictly on the presence of a well-defined NaCl concentration (100 mM). Our strain is constitutive for the enzymatic systems responsible for the oxidation of alkanes to fatty acids, i.e. NADH-dependent hydroxylase, alcohol and aldehyde dehydrogenases, the latter of which located at the cytoplasmic membrane level. The aerobic oxidation of primary alcohols by particulate extracts prepared in the presence of 400 mM NaCl is NAD+-dependent (Km = 0.082 mM, Vmax = 238 with decanol). With extracts prepared in the absence of NaCl, Vmax undergoes a very strong decrease. On the contrary , the NAD+ (P)+-dependent oxidation of aldehydes is carried out anaerobically by the same extracts irrespective of the presence or the absence of added Na+ in the solutions used for the preparation of these extracts. A possible explanation for our results could be that Na+ acts on the enzymatic systems for which the maintenance of the membrane integrity is essential. This interpretation is consistent with the slowing down of the growth speed accompanying the decrease of NaCl concentration in the growth medium. With regard to alcohol and aldehyde-dehydrogenases, it is noteworthy that these enzymes behave like similar enzymatic activities induced by alkanes in other microorganisms.     

ATP hydrolysis in a marine bacterium.

The membrane-bound adenosine triphosphatase of marine pseudomonad B-16, when solubilized, is able to rebind to depleted membrane residues of the bacterium and to those of Escherichia coli.     

Cation interactions and biochemical composition of the cell envelope of a marine bacterium

Envelopes of a marine isolate, c-A1, and of a terrestrial isolate, 121, were compared for their susceptibility to disintegration in distilled water after exposure to 0.05 m MgCl(2) and to 0.1 and 1.0 m NaCl. After exposure to MgCl(2) alone, both types of envelopes remained intact in distilled water. Envelopes of marine isolate c-A1, but not of the terrestrial isolate, fragmented in distilled water after exposure to 1.0 m NaCl. Partial reaggregation of the c-A1 envelope fragments occurred on addition of MgCl(2). In cation-exchange experiments, bound Mg(++) in the envelopes of both organisms was displaced by Na(+). The envelopes of c-A1 were found to contain lipopolysaccharide, muramic acid, and a variety of phospholipids, of which the major component was phosphatidylethanolamine, accompanied by lesser amounts of phosphatidic acid, diphosphatidylglycerol, and phosphatidylserine. Analyses of envelope acid hydrolysates revealed a similar amino acid distribution in the marine and terrestrial isolates, but envelopes of c-A1 had less than half the total amino acid content of envelopes of 121 per envelope dry weight. Possible relationships between cations and biochemical components of the envelopes are considered in terms of differences in behavior of the two organisms in low ionic environments.     

Total energy flux in a marine bioluminescent bacterium

Abstract The total heat production from luminous bacteria growing on glycerol as a sole source of carbon and energy was measured in an isothermal microcalorimeter and compared to the light emitted and growth yield. Substrate concentration was measured before and after growth to calculate the amount of substrate utilized. From these data the total energy flux was determined for Vibrio harveyi : a 25 ml culture consumed 328 micromoles of glycerol (130 cal) and produced 54 calories of heat, 76.3 cal of biomass, and 3.5 millicalories of light. Bioluminescence was then only a very small fraction (0.007%) of the total energy expended.     

Sodium ion-proton antiport in a marine bacterium.

Alteromonas haloplanktis ejected protons in response to a brief respiratory pulse; the rate of decay of the resulting pH change was accelerated when Na+ was present in the suspension medium. The addition of an anaerobic NaCl solution to an essentially Na+-free anaerobic bacterial suspension induced the acidification of the suspension medium. These results and others discussed provide substantial evidence for the existence of an Na+-H+ antiporter in this organism.     

Biofilm development and cell death in the marine bacterium Pseudoalteromonas tunicata

The newly described green-pigmented bacterium Pseudoalteromonas tunicata (D2) produces target-specific inhibitory compounds against bacteria, algae, fungi, and invertebrate larvae and is frequently found in association with living surfaces in the marine environment. As part of our studies on the ecology of P. tunicata and its interaction with marine surfaces, we examined the ability of P. tunicata to form biofilms under continuous culture conditions within the laboratory. P. tunicata biofilms exhibited a characteristic architecture consisting of differentiated microcolonies surrounded by water channels. Remarkably, we observed a repeatable pattern of cell death during biofilm development of P. tunicata, similar to that recently reported for biofilms of Pseudomonas aeruginosa (J. S. Webb et al., J. Bacteriol. 185:4585-4595, 2003). Killing and lysis occurred inside microcolonies, apparently resulting in the formation of voids within these structures. A subpopulation of viable cells was always observed within the regions of killing in the biofilm. Moreover, extensive killing in mature biofilms appeared to result in detachment of the biofilm from the substratum. A novel 190-kDa autotoxic protein produced by P. tunicata, designated AlpP, was found to be involved in this biofilm killing and detachment. A Delta alpP mutant derivative of P. tunicata was generated, and this mutant did not show cell death during biofilm development. We propose that AlpP-mediated cell death plays an important role in the multicellular biofilm development of P. tunicata and subsequent dispersal of surviving cells within the marine environment.     

Properties of lactate dehydrogenase in a psychrophilic marine bacterium

Lactate dehydrogenase (EC 1.1.1.27) from Vibrio marinus MP-1 was purified 15-fold and ammonium activated. The optimum pH for pyruvate reduction was 7.4. Maximum lactate dehydrogenase activity occurred at 10 to 15 degrees C, and none occurred at 40 degrees C. The crude-extract enzyme was stable between 15 and 20 degrees C and lost 50% of its activity after 60 min at 45 degrees C. The partially purified enzyme was stable between 8 and 15 degrees C and lost 50% of its activity after 60 min at 30 degrees C. The thermal stability of lactate dehydrogenase was increased by mercaptoethanol, with 50% remaining activity at 42 degrees C.     

A gas-producing marine bacterium

A gram-negative, aerogenic fermentative, marine bacterium is described and illustrated.     

Specific sodium dependence of a nitrate reductase in a marine bacterium

Abstract The two key enzymes of denitrification, the nitrate and nitrate reductases, were studied in a marine organism, Pseudomonas nautica , strain 617. Both enzymes were grown under anaerobic conditions with either nitrate or nitrous oxide as electron acceptor. The effect of sodium on these enzymes was studied. Only sodium activated the membranous and purified nitrate reductase, and none of the salts affected nitrite reductase.     

Interaction of respiration and luminescence in a common marine bacterium

Investigators have proposed for some time that bacterial luciferase forms a shunt around the pathway of respiratory electron transport. Certain physiologic evidence for coupling between luminescence and respiration has supported such a view. In this study, Vibrio harveyi cells were monitored for luminescent responses to artificial manipulation of respiratory electron flow. The effects of cyanide under aerobic and anaerobic conditions confirmed that luminescence and respiration compete for oxygen. The effects of an uncoupler of oxidative phosphorylation indicated that luminescence and respiration compete for a common reductant. Treatment with uncoupler also induced aldehyde deficiency in vivo.Abbreviations CCCP carbonyl cyanide m-chlorophenylhydrazone - Tris tris(hydroxymethyl) aminomethane     

High density cell culture of a marine photosynthetic bacterium Rhodovulum sp. with self-flocculated cells

High density cell cultivation of a marine photosynthetic bacterium, Rhodovulum sp. PS88 with self-flocculated cells was established by using a single-tower fermenter. High density cell culture with continuous cultivation was yielded 43 g dry matter l–1 with acetate as a substrate consumed at 22.5 g/l day. © Rapid Science Ltd. 1998     

The smallest ssDNA phage infecting a marine bacterium

In the marine environment, only a few lytic single-stranded DNA (ssDNA) phages have been isolated and characterized, despite the fact that diverse ssDNA bacteriophages have been discovered via metagenomic studies. In this study, we isolated and characterized a new ssDNA phage, vB_RpoMi-Mini, which infects a marine bacterium Ruegeria pomeroyi DSS-3. With a genome size of 4248 bp and only four putative open reading frames (ORF), vB_RpoMi-Mini becomes the smallest ssDNA phage among the known ssDNA phage isolates and represents the DNA bacteriophage with the least number of ORFs. Genome-wide analysis reveals that bacteriophage Mini is distantly related to the known ssDNA phages and belongs to an unclassified ssDNA phage within the Microviridae family. The presence of peptidase in vB_RpoMi-Mini genome further implies that horizontal gene transfer could be an important driving force in the evolution of ssDNA phages. Bacteriophage Mini seems to have lost the spike protein commonly seen in ssDNA phages, suggesting that ssDNA phage can be more diverse than previously thought. Metagenomic analysis indicates that Mini-like phages are widely distributed in the environments. The discovery of vB_RpoMi-Mini expands our understanding of ssDNA phages in nature, and also indicates our dearth of knowledge regarding of ssDNA phages.     

Novel epibiotic thiothrix bacterium on a marine amphipod

Comparative analysis of the 16S rRNA gene and fluorescent in situ hybridization (FISH) was used to identify epibiotic filamentous bacteria living on the marine amphipod crustacean Urothoe poseidonis. The epibionts belong to the gamma proteobacteria and represent a novel marine phylotype within the genus Thiothrix. FISH and denaturing gradient gel electrophoresis revealed that the Thiothrix filaments are present on the majority of the amphipods examined.