Effects of lipids on n-alkane attenuation in media supporting oil-utilizing microorganisms from the oily Arabian Gulf coasts

The Arabian Gulf is one of the most extensively oil-polluted areas of the world. The major objectives of this work were to study whether hydrocarbon-utilizing microorganisms indigenous to that area would readily accumulate added lipids, and whether this might affect their hydrocarbon consumption potential. Two prokaryotes, Arthrobacter nicotianae KCC B35 and the unidentified organisms KCC B6, as well as one eukaryote, Candida parapsilosis KCC Y1, were selected for this study. Biomass samples of the test organisms were incubated in an inorganic medium containing various concentrations of cholesterol, stearic acid, triolein or egg-phospholipids. The results revealed that all lipid classes were readily accumulated by the three test organisms. In addition, biomass samples were incubated for 6 h in an inorganic medium containing mixtures of individual lipid classes and either n-octadecane or n-docosane. The cells were removed and the residual alkanes in the medium were quantitatively recovered and analyzed by GLC. The results showed that out of the tested lipid classes only stearic acid exhibited a common stimulatory effect on the consumption of both n-alkanes by all test organisms. Other lipid classes were either inhibitory or had less pronounced effects than stearic acid.     

Enzyme polymorphisms in the mullet,Liza dussumeiri from Shatt Al-Arab river Khor Al-Zubair and the Arabian gulf

Samples of the muscles and other tissues of Lisa dussumeiri were collected from the Shatt al-Arab river, Khor al-Zubair and Kuwait (Arabian Gulf) and subjected to starch gel electrophoretic analysis of two enzymes-coding loci. All loci at all localities were in Hardy-Weinberg equilibrium. Allele frequencies at the two loci were remarkably constant throughout the area studied. The data provide no evidence of subpopulation differentiation and there is one stock of L. dussumeiri in the area studied.     

Primary distribution of n-alkane through the structures of yeast cells]

When n-alkane enters the cells as a result of molecular sorption and diffusion, it is distributed within all membrane structures of the cell (microsomes, mitochondria, membranes, vacuoles, cytoplasmic membranes, and cell walls). Sorption of n-alkane by all these structures in vivo comes at equilibrium after incubation of the cells with n-alkane during 3-4 min. Accumulation of the hydrocarbon in the morphological fractions of the cell depends on its concentration in the incubation medium. Isotherms of the sorption are convex curves. Sorption power, maximum sorption capacity, affinity and strength of the bonds with the hydrocarbon differ among the membrane structures of the cell. The maximum capacity of sorption of n-alkane by the structures does not correlate with the content of lipids and phospholipids in the structures. Sorption of n-alkanes is presumed to depend on the structural organization of lipids in the morphological fractions.     

Halophilic Nuclease from a Moderately Halophilic Micrococcus varians

The moderately halophilic bacterium Micrococcus varians, isolated from soy sauce mash, produced extracellular nuclease when cultivated aerobically in media containing 1 to 4 M NaCl or KCl. The enzyme, purified to an electrophoretically homogeneous state, had both ribonuclease and deoxyribonuclease activities. The nuclease had maximal activity in the presence of 2.9 M NaCl or 2.1 M KCl at 40 C. The enzymatic activity was lost by dialysis against low-salt buffer, whereas when the inactivated enzyme was dialyzed against 3.4 M NaCl buffer as much as 77% of the initial activity could be restored.     

Zeta-potential of n-alkane emulsion droplets and its role in substrate transport into yeast cells

Zeta-potential of n-alkane droplets, formed by fatty acids, were studied in model systems of culture liquid of yeasts (Candida maltosa) capable of utilizing n-alkanes. The value of zeta-potential was found to depend on the droplet size. The negative zeta-potential of submicron droplets was so high that it prevented the droplets from being coagulated with cells possessing a high negative zeta-potential. The dominant role of submicron n-alkane droplets in the kinetics of yeast growth could be accounted for by the existence of a mechanism regulating contact interactions of individual cells with the droplets followed by uptake of the substrate.     

Halophilic amylase from a moderately halophilic Micrococcus

A moderately halophilic Micrococcus sp., isolated from unrefined solar salt, produced a considerable amount of extracellular dextrinogenic amylase when cultivated aerobically in media containing 1 to 3 m NaCl. The Micrococcus amylase had maximal activity at pH 6 to 7 in 1.4 to 2 m NaCl or KCl at 50 C. Calcium ion and a high concentration of NaCl or KCl were essential for activity and stability of the amylase. The salt response of the amylase depended greatly on the pH and temperature of the enzyme assay.     

酵母天然酶系生物合成谷胱甘肽

利用酵母细胞本身具有的谷胱甘肽（GSH）合成酶和ATP合成GSH。在含有半胱氨酸、谷氨酸、甘氨酸的反应液中,加入已充分洗涤的酵母细胞,37℃孵育8h后,酵母体内可积累15mg/g(dry cell weight)以上的GSH,而对照组仅5mg/g(dry cell weight)。同时也研究了树脂法和铜盐法相结合分离纯化GSH。实验结果初步表明,利用酵母细胞中的天然谷胱甘肽合成酶和ATP,可使前体物质快速转化成GSH,是可行的。     

The substrate specificity of two yeast strains utilizing hydrocarbons

StrainsCandida lipolytica 4-1 andCandida lipolytica K were compared in their growth and dawaxing capacities during batch growth on model gas oil. The model gas oil was composed of a mixture of even-numbered puren-alkanes (n-decane ton-dotriacontane) dissolved in dewaxed gas oil. The results show that both strains differ in their substrate specificity and in the sequence of utilization of individualn-alkanes. Strain K, previously used for dewaxing of mineral oil, has its substrate specificity shifted toward the highern-alkanes.     

First dinosaur tracks from the Arabian Peninsula

Background

The evolutionary history of Mesozoic terrestrial vertebrates from the Arabian Peninsula is virtually unknown. Despite vast exposures of rocky outcrops, only a handful of fossils have yet been described from the region. Here we report a multi-taxon dinosaur track assemblage near Madar village, 47 km north of Sana''a, Republic of Yemen. This represents the first dinosaur tracksite from the Arabian Peninsula, and the only multi-taxon dinosaur ichnosite in the Middle East.

Methodology/Findings

Measurements were taken directly from trackway impressions, following standard ichnological conventions. The presence of bipedal trackmakers is evidenced by a long series of pes imprints preserving smoothly rounded posterior margins, no evidence of a hallux, bluntly rounded digit tips and digital divarication angles characteristic of ornithopod dinosaurs. Nearby, eleven parallel quadrupedal trackways document a sauropod herd that included large and small individuals traveling together. Based on the morphology of manus impressions along with a narrow-gauged stance, the quadrupedal trackways were made by non-titanosauriform neosauropods. Additional isolated tracks and trackways of sauropod and ornithopod dinosaurs are preserved nearby.

Conclusions/Significance

Taken together, these discoveries present the most evocative window to date into the evolutionary history of dinosaurs of the Arabian Peninsula. Given the limited Mesozoic terrestrial record from the region, this discovery is of both temporal and geographic significance, and massive exposures of similarly-aged outcrops nearby offer great promise for future discoveries.     

Halophilic and halotolerant aerobic methylobacteria from the technogenic Solikamsk biotopes

Seven strains of moderately halophilic and halotolerant aerobic methylobacteria from the technogenic Solikamsk biotopes (Perm krai, Russia) were isolated in pure cultures and characterized. The isolates were represented by gram-negative and gram-positive (strain 2395B) cells. All the cells were shown to multiply by binary fission without formation of spores or prosthecae. All isolates except strain 2395B were able to oxidize methanol by a classical methanol dehydrogenase. The ribulose monophosphate (RMP) (strain LS), serine (strains S12, S3, 2395A), or ribulose bisphosphate (strains SK15 and S3270) pathways of C₁-assimilation were used. In strain 2395B, the key enzymes of the RMP and serine metabolic pathways were determined. Using polyphasic taxonomy, three strains were identified as representatives of the known species: Arthrobacter protophormiae 2395B, Methylophaga thalassica LS, and Ancylobacter rudongensis S3270. Three more strains were identified as members of new species: Methylopila oligotropha sp. nov. (strain 2395AT; VKM B-2788^T = CCUG 63805^T), Ancylobacter defluvii sp. nov. (strain SK15^T; VKM B-2789^T = CCUG 63806^T), and Paracoccus communis sp. nov. (strain S3^T; VKM B-2787^T = CCUG 63804^T). According to the results of 16S rRNA gene sequencing, the obligately methylotrophic strain S12 had less than 94% similarity with the known genera of the Proteobacteria and was probably a representative of a novel genus.     

Rescue of a single yeast artificial chromosome from a cotransformation event utilizing segregation at meiosis

During the construction of yeast artificial chromosome (YAC) libraries to facilitate mapping of the human genome, two YACs may be cotransformed into the same yeast cell, making further analysis very difficult. We present a simple method to rescue the required YAC that utilizes the segregation of chromosomes at meiosis. In brief, we crossed the cotransformed yeast cell with a non-YAC-containing strain and induced the resulting diploid to sporulate and undergo meiosis. The new haploid generation included some yeast cells that contained only the desired YAC. These YACs were analyzed by conventional methods. To exclude the possibility that major rearrangement occurred during the procedure, we analyzed the YACs with restriction enzymes that cut only rarely. We conclude that this is a useful technique to rescue cotransformed YACs.     

Bacillusstearothermophilus from Saudi Arabian soils

Ten thermophilic Gram-positive bacteria were isolated from various soils of Saudi Arabia. The strains are spore-forming rods belonging to the speciesBacillus stearothermophilus. The cells are motile, strictly aerobic, catalase and oxidase positive. The sporangia appear to be swollen and their position varies from terminal in some to sub-terminal in others. The thermal stability of some enzymes of these bacteria was investigated; extracellular α-amylase appears to be very sensitive to pH and temperature. The ultrastructure of these bacteria shows specific changes in the cell wall when grown at the maximum, minimum and optimum growth temperatures, respectively.     

Halophilic Archaea determined from geothermal steam vent aerosols

Hydrothermal vents, known as 'fumaroles', are ubiquitous features of geothermal areas. Although their geology has been extensively characterized, little is known about the subsurface microbial ecology of fumaroles largely because of the difficulty in collecting sufficient numbers of cells from boiling steam water for DNA extraction and culture isolation. Here we describe the first collection, molecular analysis and isolation of microbes from fumarole steam waters in Russia (Kamchatka) and the USA (Hawaii, New Mexico, California and Wyoming). Surprisingly, the steam vent waters from all the fumaroles contained halophilic Archaea closely related to the Haloarcula spp. found in non-geothermal salt mats, saline soils, brine pools and salt lakes around the world. Microscopic cell counting estimated the cell dispersal rate at approximately 1.6 x 10(9) cells year(-1) from a single fumarole. We also managed to enrich microbes in high-salt media from every vent sample, and to isolate Haloarcula from a Yellowstone vent in a 20% salt medium after a month-long incubation, demonstrating both salt tolerance and viability of cells collected from high-temperature steam. Laboratory tests determined that microbes enriched in salt media survived temperatures greater than 75 degrees C for between 5 and 30 min during the collection process. Hawaiian fumaroles proved to contain the greatest diversity of halophilic Archaea with four new lineages that may belong to uncultured haloarchaeal genera. This high diversity may have resulted from the leaching of salts and minerals through the highly porous volcanic rock, creating a chemically complex saline subsurface.     

Halophilic Vibrio species from seafish in Senegal.

Sucrose-positive and sucrose-negative halophilic Vibrio species at counts of up to 10(7)/100 g were isolated from muscles tissue in 27 and 43%, respectively, of 128 seafish from coastal waters in Senegal. Vibrio parahaemolyticus, including 21% urease-positive strains, was the most common isolate, followed by Vibrio alginolyticus, Vibrio vulnificus, Vibrio damsela, and Vibrio fluvialis.     

A First Record of Obligate Halophilic Aspergilli from the Dead Sea

The isolation of obligate halophilic aspergilli from the Dead Sea and the range of salt tolerance of halophilic fungi isolated, are reported here for the first time. The mycobiota of the Dead Sea isolated in this study, was dominated by Aspergillus and Penicillium species; Cladosporium were found in lesser numbers. All three genera were obtained from the water sample; however, Aspergillus was the only genus obtained from the sediment. There was significant difference in growth of each isolate at different salt concentrations and intraspecies analysis revealed dissimilarity in response of strains to different salt concentrations in the growth medium The isolates were euryhaline, with halotolerance up to 20–25% solar salt, Aspergillus and Penicillium species showing a higher level of halotolerance, as compared to that of Cladosporium. Halophilic fungi were found in greater numbers in the sediment sample as compared to that in the water sample. Penicillium and Cladosporium species were exclusively facultative halophiles, while some species of Aspergillus were facultative halophiles. All the obligate halophiles isolated, belonged to the genus Aspergillus and were identified as A. penicillioides and A unguis, the latter being a first record of the species from the Dead Sea.     

A network-based analysis of polyanion-binding proteins utilizing yeast protein arrays

The high affinity of certain cellular polyanions for many proteins (polyanion-binding proteins (PABPs)) has been demonstrated previously. It has been hypothesized that such polyanions may be involved in protein structure stabilization, stimulation of folding through chaperone-like activity, and intra- and extracellular protein transport as well as intracellular organization. The purpose of the proteomics studies reported here was to seek evidence for the idea that the nonspecific but high affinity interactions of PABPs with polyanions have a functional role in intracellular processes. Utilizing yeast protein arrays and five biotinylated cellular polyanion probes (actin, tubulin, heparin, heparan sulfate, and DNA), we identified proteins that interact with these probes and analyzed their structural and amino acid sequence requirements as well as their predicted functions in the yeast proteome. We also provide evidence for the existence of a network-like system for PABPs and their potential roles as critical hubs in intracellular behavior. This investigation takes a first step toward achieving a better understanding of the nature of polyanion-protein interactions within cells and introduces an alternative way of thinking about intracellular organization.