Biosynthesis of medium chain length poly(3-hydroxyalkanoates) (mcl PHAs) from cosmetic co-products by Pseudomonas raguenesii sp. nov., isolated from Tetiaroa, French Polynesia

A new bacterium, designated as strain TE9 was isolated from a microbial mat in French Polynesia and was studied for its ability to synthesize medium chain length poly-β-hydroxyalkanoates (mcl PHAs) during cultivation on cosmetics co-products. The composition of PHAs was analysed by coupled gas chromatography mass spectroscopy (GC/MS), nuclear magnetic resonance (NMR) and Fourier Transform InfraRed (FTIR) spectroscopy. PHAs were composed of C6–C14 3-hydroxyacids monomers, with a predominance of 3-hydroxyoctanoate (3HO), 3-hydroxydecanoate (3HD) and 3-hydroxydodecanoate (3HDD). Differential scanning calorimetry (DSC) experiments allowed the characterization of elastomeric materials with a melting point T_m near 50 °C, enthalpy of fusion ΔH_m from 27 to 32 J/g, and glass transition temperature T_g of −43 °C. Molecular weights ranged from 175,000 to 358,000 g/mol. On the basis of the phenotypical features and genotypic investigations, strain TE9 was assigned to the Pseudomonas genus and the name of Pseudomonas raguenesii sp. nov. is proposed.     

A novel mcl PHA-producing bacterium, Pseudomonas guezennei sp. nov., isolated from a 'kopara' mat located in Rangiroa, an atoll of French Polynesia

Aims: The aim of the present study was to describe an aerobic, mesophilic and heterotrophic bacterium, designated RA26, able to produce a medium‐chain‐length polyhydroxyalkanoate (PHA). It was isolated from a French Polynesian bacterial mat located in the atoll of Rangiroa. Methods and Results: This micro‐organism, on the basis of the phenotypical features and genotypic investigations can be clearly assigned to the Pseudomonas genus and the name of Pseudomonas guezennei is proposed. Optimal growth occurs between 33 and 37°C, at a pH between 6·4 and 7·1 and at ionic strength of 15 g l^?1 of sea salts. The G+C content of DNA is 63·2%. Under laboratory conditions, this bacterium produced a novel, medium‐chain‐length PHA, mainly composed of 3‐hydroxydecanaote (64 mol.%) and 3‐hydroxyoctanoate (24 mol.%) (GC‐MS, NMR) from a single nonrelated carbon substrate, i.e. glucose. Conclusions: The bacterium P. guezennei produces a novel PHA mcl with elastomeric properties. Significance and Impact of the Study: PHAs share physical and material properties that recommend them for application in various areas, and are considered as an alternative to nonbiodegradable plastics produced from fossil oils. In this study, we describe a new bacteria with the capability to synthesize a novel PHA with promising biotechnological applications.     

A single-cell sequencing approach to the classification of large, vacuolated sulfur bacteria

The colorless, large sulfur bacteria are well known because of their intriguing appearance, size and abundance in sulfidic settings. Since their discovery in 1803 these bacteria have been classified according to their conspicuous morphology. However, in microbiology the use of morphological criteria alone to predict phylogenetic relatedness has frequently proven to be misleading. Recent sequencing of a number of 16S rRNA genes of large sulfur bacteria revealed frequent inconsistencies between the morphologically determined taxonomy of genera and the genetically derived classification. Nevertheless, newly described bacteria were classified based on their morphological properties, leading to polyphyletic taxa. We performed sequencing of 16S rRNA genes and internal transcribed spacer (ITS) regions, together with detailed morphological analysis of hand-picked individuals of novel non-filamentous as well as known filamentous large sulfur bacteria, including the hitherto only partially sequenced species Thiomargarita namibiensis, Thioploca araucae and Thioploca chileae. Based on 128 nearly full-length 16S rRNA-ITS sequences, we propose the retention of the family Beggiatoaceae for the genera closely related to Beggiatoa, as opposed to the recently suggested fusion of all colorless sulfur bacteria into one family, the Thiotrichaceae. Furthermore, we propose the addition of nine Candidatus species along with seven new Candidatus genera to the family Beggiatoaceae. The extended family Beggiatoaceae thus remains monophyletic and is phylogenetically clearly separated from other related families.     

A bacterial extracellular proteinase degrading silk fibroin

The bacterium Variovorax paradoxus, grown in a minimal medium in which silk fibroin represents the sole source of carbon and nitrogen, produces an extracellular protease that hydrolyzes fibroin as well as casein and, to a smaller extent, collagen and albumin. The optimal pH for activity was found to be in the acid range (optimum pH 5.8–6.4) and the enzyme activity was stimulated by the addition of divalent cations, either manganese or magnesium. Gel permeation chromatography and SDS-PAGE provided evidence that the native enzyme is a monomer with a M_r of ca. 21 kDa.     

Several pseudomonads, associated with the cultivated mushrooms Agaricus bisporus or Pleurotus sp., are hemolytic

Pseudomonas tolaasii, causing brown blotch disease on cultivated mushrooms, and yielding a white line precipitate towards P. “reactans”, has been shown to induce lysis of erythrocytes. Some Finnish strains isolated from diseased mushroom fruit bodies, although harboring the typical features of P. tolaasii, proved to be distinct, and have been allocated to a nov. sp. P. costantinii. We examined in these study whether all brown blotch causing agents were hemolytic. The induction of erythrocytes lysis seemed to be a rather common feature of mushroom associated-pseudomonads, especially for strains involved in the production of a white-line-in agar.     

Degraded temperature sensitivity of a stone pine chronology explained by dendrochemical evidences

Fluctuations in sulphur (S) content in tree rings were analysed for a 93 years long period (1915–2007) by laser ablation system coupled to an inductively coupled plasma mass spectrometer from three Swiss stone pine (Pinus cembra L.) trees grown in the Rachitis Cirque, Calimani Mts, Eastern Carpathians. Investigated domain envelops the period of degraded climatic sensitivity of pines’ growth. Chemical data were standardized for sapwood and heartwood separately. The averaged sulphur record calculated from the three individual records unequivocally presents anomalously high S content for the tree rings dated to 1970s and 1980s. Ring-width fluctuation portrays changes of summer temperature but from 1966 to 1986 pines produced apparently larger rings than the corresponding summer temperatures can explain. This deviating interval coincides with the period of exploitation in the nearby sulphur mine. The elevated S content in tree rings refers probably to the dispersion of S-rich dust around the mine owing to the opencast type of exploitation. The divergent growth trend and degraded climatic signal likely due to the anthropogenic altered nutritional status of the site, as residues from nitrogen fertilizer, applied as a compound of explosive in the opencast sulphur exploitation, could be deposited on the surrounding forest.     

Arsukibacterium ikkense gen. nov., sp. nov, a novel alkaliphilic, enzyme-producing gamma-Proteobacterium isolated from a cold and alkaline environment in Greenland

A novel aerobic, Gram-negative, non-pigmented bacterium, GCM72(T), was isolated from the alkaline, low-saline ikaite columns in the Ikka Fjord, SW Greenland. Strain GCM72(T) is a motile, non-pigmented, amylase- and protease-producing, oxidase-positive, and catalase-negative bacterium, showing optimal growth at pH 9.2-10.0, at 15 degrees C, and at 3% (w/v) NaCl. Major fatty acids were C(12:0) 3-OH (12.2+/-0.1%), C(16:00) (18.0+/-0.1%), C(18:1)omega7c (10.7+/-0.5%), and summed feature 3 comprising C(16:1)omega7c and/or iso-C(15:0) 2-OH (36.3+/-0.7%). Phylogenetic analysis based on 16S rRNA gene sequences showed that isolate GCM72(T) was most closely related to Rheinheimera baltica and Alishewanella fetalis of the gamma-Proteobacteria with a 93% sequence similarity to both. The G+C content of DNA isolated from GCM72(T) was 49.9mol% and DNA-DNA hybridization between GCM72T and R. baltica was 9.5%. Fatty acid analysis and G+C content supports a relationship primarily to R. baltica, but several different features, such as a negative catalase-response and optimal growth at low temperature and high pH, together with the large phylogenetic distance and low DNA similarity to its closest relatives, lead us to propose a new genus, Arsukibacterium, gen. nov., with the new species Arsukibacterium ikkense sp. nov. (type strain is GCM72(T)).     

Photosynthetic characteristics of the economic brown seaweed <Emphasis Type="BoldItalic">Hizikia fusiforme</Emphasis> (Sargassaceae,Phaeophyta), with special reference to its “leaf” and receptacle

Photosynthetic responses to irradiance and temperature of “leaves” and receptacles were compared in February (vegetative stage) and May (reproductive stage) in the seaweed, Hizikia fusiforme (Harvey) Okamura (Sargassaceae, Phaeophyta) from Nanao Island, Shantou, China. Irradiance-saturated photosynthesis (P_max) was significantly higher in receptacles than in “leaves” on a fresh weight basis, and that of “leaves” was greater in May than in February at ambient seawater temperatures. The optimum temperature for P_max was 30^∘C for both “leaves” and receptacles, being 5–10^∘C higher than the ambient seawater temperature. The apparent photosynthetic efficiencies were greater in receptacles than in “leaves” within the tested temperature range of 10–40^∘C. The irradiance for saturating photosynthesis for both “leaves” and receptacles was temperature-dependent, with the highest values (about 200μmolphotonsm⁻²s⁻¹) at 30^∘C.     

Desulfurization of light gas oil by a novel recombinant strain from Pseudomonas aeruginosa

The dsz desulfurization gene cluster from Rhodococcus erythropolis KA2-5-1 was transferred into the chromosomes of Pseudomonas aeruginosa NCIMB 9571 by using a transposon vector. Resting cells of the recombinant strain, PAR41, desulfurized 63 mg sulfur l^–1 of light gas oil (LGO) containing 360 mg S l^–1. The desulfurization activity for LGO by the resting cells of strain PAR41 grown with n-tetradecane (50% v/v) was much higher (1018-fold) than in glucose-grown cells. P. aeruginosa NCIMB 9571 is able to take up water-insoluble compounds from an oil phase which is enhanced by n-alkane.     

The Casal de' Pazzi archaic parietal: comparative analysis of new fossil evidence from the late Middle Pleistocene of Rome

A fossilized fragment of human parietal bone has been recently recovered from the lowest layer of the Casal de' Pazzi fluvial deposit (stratigraphically dated at about 200–250 ky BP). The fossil presents characters-i.e., thickness, degree and development of curvature, type of endocranial vascularization-which distinguish it from the corresponding cranial regions of both Homo erectus and anatomically modern Homo sapiens. While a morphological orientation towards Neanderthal characters can be considered, the affinities of the Casal de' Pazzi parietal are primarily with other late Middle Pleistocene specimens. The authors conclude that the Casal de' Pazzi human find can be assigned to the “archaic Homo sapiens” group falling within the European pre-Neanderthal range. Its particular morphology constitutes new evidence of human evolution from the geographical area of Rome.     

Symbiotic effectiveness of strains of Rhizobium leguminosarum biovar phaseoli isolated from soils of Rwanda

We have isolated 48 strains of Rhizobium leguminosarum biovar phaseoli from nodules of Phaseolus vulgaris L. cultivated on 32 different soils at 22 various locations in Rwanda, Central Africa. The symbiotic effectiveness of the strains was appraised in the greenhouse by measuring shoots dry matter and total plant nitrogen content after six weeks of growth. Of the strains tested 19%, 58% and 23% were rated very effective, effective and ineffective, respectively. A very significant correlation (r=0.96, P<0.01) was observed between shoots dry matter and total N content. By using the total nitrogen balance method, it was estimated that in the presence of a very effective strain, up to 86% of the N present in the shoots comes from N₂ fixation. No significant correlations were observed between the symbiotic effectiveness of the strains and the pH of the soils from which they originated, the tolerance of the strains to acidity or their ability to produce organic acids. The nine very effective strains selected were highly competitive against two ineffective strains with the two P. vulgaris cultivars Rubona-5 and Kiryumukwe.Contribution no 367, Station de recherches, Agriculture Canada.Contribution no 367, Station de recherches, Agriculture Canada.     

Cloning and expression of a lipoxygenase from <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> 42A2

In order to produce (S) 10-monohydroxy-8E-octadecenoic acid (MHOD) from oleic acid, a full-length probable lipoxygenase cDNA from Pseudomonas aeruginosa 42A2 was cloned and expressed in Escherichia coli BL21(DE3). The recombinant protein was purified by affinity chromatography to electrophoretic homogeneity and specifically stained. Its molecular mass was 70 kDa. The activity of the rec-LOX with oleic acid was about 30% of that of the prefered substrate, linoleic acid (100%). Bacterial LOX forms a new subfamily in the lipoxygenase phylogenetic tree.     

Preliminary Observations on the Nature,Taxonomic Composition,Distribution, Behaviour and Nutrient Requirements of the “Unusual” Algal “Jellies” of Oguta Lake in Southeastern Nigeria

Preliminary observations were made between October 1982 and May 1983 on the nature, taxonomic composition, spatial distribution, buoyancy behaviour and nutrient requirements of some unusual planktonic algal “jellies” in Oguta Lake in Southeastern Nigeria. These “jellies” are macroscopic, blue-green algal masses whose excessive mucilage of high consistency acts as substrata for other epiphytic components viz. diatoms and green algae. They are more abundant in the lentic areas of the lake, undergo diurnal vertical movements and have their growth enhanced by spikes of nitrogen and phosphorus salts into their aquatic medium. The effects of these “jellies” on the ecology and general water use of the lake are discussed. Speculations on the reasons for their occurrence are given and possible control measures outlined.     

Planococcus rifietensis sp. nov, isolated from algal mat collected from a sulfurous spring in Campania (Italy)

The taxomony of strain M8, isolated from algal mat formed at the origin of a sulfurous spring in Rifieto (Savignano Irpino, Campania, Italy), was investigated in a polyphasic approach. The morphological, physiological and genetic characteristics were compared with of Planococcus and Planomicrobium species. The isolate grew optimally at pH 9.0, 1.8 M NaCl at 37 degrees C. The cells were Gram-positive cocci that form pairs, tetrads and aggregates of several cells. The isolate was aerobic/microaerophilic and accumulated glycine-betaine, as a major osmolyte, with minor components glutamate and an unknown compound. M8 was able to hydrolyse X-Glc (5-bromo-4-chloro-3-indoyl beta-d-glucopyranoside). The polar lipid profile consisted of phosphatidylglycerol and diphosphatidylglycerol as major components, and phosphocholine as a minor compound. MK8 was the only quinone found and the fatty acid composition was dominated by branched acids, mainly aiC15:0. The G+C content of DNA was 47.9% and its phylogenetic position was established by 16S rRNA gene sequencing as a member of the genus Planococcus. The DNA/DNA similarity of M8 to the type species Planococcus citreus was less than 55%. For this reason and for physiological and chemotaxonomic features, it is proposed to create a new species Planococcus rifietensis sp. nov.     

Growth promotion of maize by phosphate-solubilizing bacteria isolated from composts and macrofauna

Five bacterial strains with phosphate-solubilizing ability and other plant growth promoting traits increased the plant biomass (20-40%) by paper towel method. Glasshouse and field experiments were conducted using two efficient strains Serratia marcescens EB 67 and Pseudomonas sp. CDB 35. Increase in plant biomass (dry weight) was 99% with EB 67 and 94% with CDB 35 under glasshouse conditions. Increase in plant biomass at 48 and 96 days after sowing was 66% and 50% with EB 67 and 51% and 18% with CDB 35 under field conditions. Seed treatment with EB 67 and CDB 35 increased the grain yield of field-grown maize by 85% and 64% compared to the uninoculated control. Population of EB 67 and CDB 35 were traced back from the rhizosphere of maize on buffered rock phosphate (RP) medium and both the strains survived up to 96 days after sowing.     

Effect of catabolic plasmids on physiological parameters and efficiency of oil destruction by Pseudomonas bacteria

The ability of microbial degraders of polycyclic aromatic hydrocarbons to grow at 24°C in liquid mineral medium supplemented with oil as the sole source of carbon and energy was studied. Growth characteristics (CFU) and the level of oil destruction by plasmid-bearing and plasmid-free strains were determined after seven days of cultivation. The presence of catabolic plasmids in the degrader strains, including rhizosphere pseudomonads, was shown to increase cell growth and enhance the level of oil degradation. Strain Pseudomonas chlororaphis BS1391 bearing plasmid pBS216 was found to be the most effective oil degrader.     

Metabolism of benzalphthalide by Pseudomonas sp.

A Pseudomonas sp. degraded benzalphthalide to o-phthalate and benzoate. A tentative pathway for the metabolism of benzalphthalide in this Pseudomonas sp. is proposed on the basis of isolated metabolites, oxygraphic assay and enzymatic studies.     

Cloning,expression and characterization of a beta-agarase gene from a marine bacterium,Pseudomonas sp. SK38

A gene (pagA) encoding -agarase from Pseudomonas sp. SK38 was cloned and expressed in Escherichia coli. The structural gene consists of 1011 bp encoding 337 amino acids with a predicted molecular weight of 37326 and has a signal peptide of 18 amino acids. The deduced amino acid sequence showed 57% and 58% homology to -agarase from Pseudoalteromonas atalntica and Aeromonas sp., respectively. The recombinant enzyme was purified and biochemically characterized. The enzyme had maximum activity at pH 9 and 30 °C. It was stable at pHs from 8 to 9 and below 37 °C.