Characterization of two halophilic adenylate cyclases from Thermobifida halotolerans and Haloactinopolyspora alba

Adenylate cyclase(AC)is the key enzyme that catalyzes the formation of cAMP from ATP.In this study,we discovered two novel class Ⅲ ACs with a halophilic property from Thermobifida halotolerans DSM 44931(ThAC)and Haloactinopolyspora alba DSM 45211(HaAC),respectively.The recombinant ThAC and HaAC were expressed in Escherichia coli with molecular weights of 36.1 and 36.0 kDa respectively.The presence of 2500 and 2200 mmol
L^(-1)1 NaCl significantly enhanced the enzyme activities of ThAC and HaAC,with 22-fold and 7.4-fold higher activities compared to those without NaCl,respectively.Several divalent metal ions were found to activate the recombinant ACs to different extents,and the optimal metal ion was Mg^(2+)for both ThAC and HaAC with concentrations of 80 mmol·L^(-1) and 40 mmol·L^(-1) respectively.Purified ThAC and HaAC had the optimal specific activities((4.59±0.35)×10^(4) and(7.76±0.52)×10^(4) U·mg^(-1))and catalytic efficiency(4.47 and 5.30 L·mmol^(-1)·s^(-1))at 45
℃ and 40
℃ respectively,while the optimum pH of both two recombinant ACs was 10.0.This is the first report of the halophilic Class III ACs,which could make new contributions to explore and study ACs for further associated investigations.

Isolation and characterization of halophilic Bacillus sp.BS3 able to produce pharmacologically important biosurfactants

Objective:To characterize the pharmacological importance of biosurfactants isolated from halophilic Bacillus sp BS3.Methods:Halophilic Bacillus sp.BS3 was isolated from solar salt works,identified by 16S rRNA sequencing and was used for screening their biosurfactant production.Characters of the biosurfactant and their anticancer activity were analyzed and performed in mammary epithelial carcinoma cell at different concentrations.Results:The biosurfactant were characterized by TLC,FTIR and GC-MS analysis and identified as lipopeptide type.GC-MS analysis revealed that,the biosurfactant had various compounds including 13Docosenamide.(Z);Mannosamine,9- and N,N,N',N'-tetramethyl.Surprisingly the antiviral activity was found against shrimp white spot syndrome virus(WSSV) by suppressing the viral replication and significantly raised shrimp survival(P<0.01).Anticancer activity performed in the mammary epithelial carcinoma cell at different concentrations of biosurfactants,among the various concentrations of biosurfactants such as 0.000 25,0.002 5,0.025,0.25 and 2.5 μ g,the 0.25 μ g concentration suppressed the cells significantly(P<0.05) to 24.8%.Conclusions:Based on the findings,the present study concluded that,there is a possibility to develop eco-friendly antimicrobial and anticancer drugs from the extremophilic origin.

Floristic composition of the halophilic and salt-resistant plant population in Hammam-Boughrara (Oran-Algeria)

This phyto-ecological study is on halophilic and salt-resistant vegetation of Oran region. The semiarid and sometimes arid climate has been defined and confirmed from a bioclimatic point of view. The pedological approach used shows a soil with sandy to silty-sandy texture, favoring regression of the vegetation and a halophilic vegetation set up. In this study, we analyze the floristic composition of the northern region of Hammam Boughrara using multiple floristic surveys conducted at three stations along the Tafna wadi. Dominated by Mediterranean and Saharo-Sindian elements, the relatively poor flora (88 species in total) is biologically characterized by a clear dominance of therophytes (>33%) and chamaephytes (>19%) to the detriment of phanerophytes.

Halophilic Vibrios in the waters of Xiamen Harbor and Daya Bay

The distribution featires, spotes composition and seasonal variation of halophilic Vibrios in 7 stations ofXiamen Harbor and 9 stations of the Daya Bay were studied. The counts of Vibrios were analyzed with the most probable number(MPN) technique. The media TCBS was used to isolate Vibrios, and API20E system employed to identifythe Vibrios. The results reveals that the density Of Vibrios in the Daya Bay ranged from 30. 0 x 105 to 2. 4 x 105 celldin-3 and the average density was 6. 61 x 103 cell·din-3, and that in Xiamen Harbor was 2. 3 x 102-2. 4 x 105 cen·din-3 and 7. 8 x 103 cell·din-3 on the average. The number of Vibrios varied seasonally with the water temperature,and was heher in the summer than in the autumn. The Vibrio species in the two bays mainly included Vibrioalginolyticus, V. parahaemolytic-us, V. fluvialis, V. vulnificus, V. mimices and V. metshnikovii and V.alginolyticus was the predominant spotes.

Biodiversity and Screening of Moderately Halophilic Bacteria with Hydrolytic and Antimicrobial Activities from Yuncheng Salt Lake,China

1 Introduction Yuncheng Salt Lake is an old lake with a history of more than 5000 years,which locates in the margins of the central plains of China,with an area of about 130 km.It was an important resource of salt for human consumption from early times.Major chemicals in Yuncheng Salt Lake contain Na Cl,Na2SO4 and Mg SO4(Gao et al.2007).In recent years,some halophilic bacteria and archaea were

Effect of salinity on community structure and naphthalene dioxygenase gene diversity of a halophilic bacterial consortium

The aim of this study is to analyze the effect of salinity on polycyclic aromatic hydrocarbons （PAHs） biodegradation, community structure and naphthalene dioxygenase gene （ndo） diversity of a halophilic bacterial consortium with the denaturing gradient gel electrophoresis （DGGE） approach. The consortium was developed from oil-contaminated saline soil after enrichment for six times, using phenanthrene as the substrate. The prominent species in the bacterial consortium at all salinities were identified as halophilic bacteria Halomonas, Alcanivorax, Marinobacter, Idiomarina, Martelella and uncultured bacteria. The predominant microbes gradually changed associating with the saline concentration fluctuations ranging from 0.1% to 25% （w/v）. Two ndo alpha subunits were dominant at salinities ranging, from 0.1% to 20%. while not been clearly detected at 25% salinity. Consistently. the biodegradation occurred at salmltles rangmg from 0.1% to 20%, while no at 25% salinity, suggesting the two ndo genes played an important role in the degradation. The phylogenetic analysis revealed that both of the two ndo alpha subunits were related to the classic nab-like gene from Pseudomonas stutzeri AN10 and Pseudomonas aeruginosa PaK1, while one with identity of about 82% and the other one with identity of 90% at amino acid sequence level. We concluded that salinity greatly affected halophilic bacterial community structure and also the functional genes which were more related to biodegradation.

Genetic analysis of the br gene in halophilic archaea isolated from Xinjiang region, China

Some novel members of extremely halophilic archaea,strains AJ11,AJ12 and AJ13,were isolated from the Aularz Lake located in the Altun Mountain National Nature Reserve of Xinjiang,Uygur Autonomous Region in China.Partial DNA fragments encoding a bacteriorhodopsin(BR),as well as for 16S rRNA of isolated strains,were amplified by PCR and their DNA sequences were determined subsequently.On the basis of homology and phylogenetic analysis of the 16S rDNA,we thought that the isolated strains forming a microbiological population are the members of the genus Natrinema.The results of genetic analysis,such as GC content,transition/transversion(Ti/Tv)rate ratios and synonymous substitution rates(Ks)indicate that the br fragments,with a high level of genetic divergence,are faced with both purifying selection and bias mutation pressure.The study provides the basis for use of species and BR proteins resources.

Screening and Denitrification Characteristics of a Slight Halophilic Heterotrophic Nitrobacteria

A slight halophilic heterotrophic nitrobacteria named gs1 was separated from the matured activated sludge. According to the morphological observation,physiological biochemical tests and sequence analysis of the 16S rDNA,strain gs1 was identified to be as Pseudomonas sp. Sodium acetate and ammonium chloride were used as carbon and nitrogen sources,respectively,to investi-gate the characteristics of the bacterium. When cultured for 24 h under aerobic conditions,with the removal rates of the NH4+-N and COD being 82.2% and 74.73%,respectively,strain gs1 will have a nitrification function of producing NO2--N. When cultured for 24 h under aerobic conditions in nitrite medium,the removal rate of the NO2--N became 100%,and when cultured for 24 h under aerobic conditions in nitrate medium,the removal rate of the NO3--N became 97%. The result shows that this strain functions for either nitrification or denitrification,i.e.,it can complete the full process of biological deoxidation.

Isolation and Phylogenetic Analysis of Halophilic Archaeon AJ6

Halophilic archaeon AJ6 was isolated and purified from the Altun Mountain National Nature Reserve of the Xinjiang Uygur Autonomous Region.Strain AJ6 is a Gram-negative rod whose size is 0.2–0.6 by 1.6–4.2μm,wherein a few cells are globular.The optimum salt concentration for its growth is 20%NaCl and 0.6%Mg^(2+),and the optimum pH is 6.0–7.0.Morphological,physiological,and biochemical characteristics of strain AJ6 were observed.The 16S rRNA encoding gene(16S rDNA)sequence of strain AJ6 was amplified by PCR,and its nucleotide sequence was determined subsequently.“Clustalw”and“PHYLIP”software bags were used to analyze the 16S rDNA sequence;the homology was compared,and then the phylogenetic tree was established.The results indicate that strain AJ6 is a novel species of the genus Natrinema.The GenBank accession number of the 16S rDNA sequences of strain AJ6 is AY277584.

Relative reactivities of halogen-substituted substrates (R-Br,R-C1) toward the halophilic attack by a carbanion

Relative reactivities of bromine-substituted substrates (R-Br) or chlorine-substituted substrates (R-CI) toward bromophilic or chlorophilic attack by a carbanion have been evaluated by the intermolecular competition kinetics. Relative reactivity orders are CF3CFBr2 >CF3CBr3≥CBr4 > CHBr3 > CF3CFBrCF2Br > CF2Br2 > BrCF2CF2Br > BrCH2CO2Et≥ BrCF2CFHBr > CH2Br2 > BrCH2CH2Br, and CI3CNO2 > CI3CCN > CI3CCOPh > cyclo-C5CI6> CI3CCOCI > CCI3CF2CI > CCI3CF3 ≥ CCI4 > CCI3CCI3 ≥ CCI3(CF2)2CI > CI3CCOCCI3 > CCI3(CF2)6CI > CI3CCO2Et > CI3CF > CI3CPh>CI3CCH2O2CCH3.

An analysis of Great Salt Lake Winogradsky columns

Sergei Winogradsky illuminated revolutionary concepts and produced a tool to visualize complex microbial communities and their metabolisms over time:columns displaying aquatic consortia with variety of niches.We worked with museums in Utah to create Winogradsky columns that would highlight aesthetic properties of the Great Salt Lake(GSL)ecosystem,which has a salinity gradient from the freshwater wetlands to salt saturation.One column,constructed using haloarchaea-rich hypersaline brine and oolitic sand of the lake’s north arm,was enriched with nutrients,and resulted in the desired pink hue over time.After a seven-year maturation period,we examined the microbial taxa present in the water through 16 S/18 S rRNA and Internal Transcribed Spacer(ITS)gene sequencing.A pigment analysis revealed an abundance of bacteriochlorophyll a.The presence of this pigment coupled with the DNA sequencing results,suggest that the haloarchaea that dominate the GSL brine,were not responsible for the pink coloration,but instead Gammaproteobacteria,especially Halorhodospira species.Among the eukaryotes,the lack of phytoplankton and the abundance of fungi were noteworthy observations.These data likely relate to the reduction of oxygen in a non-aerated sealed system over time.Our second exhibit had the goal of educating museum goers about the varying salinities of Great Salt Lake.Here we employed three distinct columns of water and sediment from this salinity gradient.Observations of these columns overtime gave us information about invertebrate communities in addition to the microbial consortia.Both installations taught us about comparing an artificial environment in a museum setting to the natural ecosystem.Taken together,we present the data collected and lessons learned from using Winogradsky columns in public spaces for teaching about an important saline lake.

A Study of the Characteristics of Microorganisms for Effective Degradation of Marine Oil Spills

Four microorganism strains were isolated from coastal petroleum-polluted soil and sand samples of Bohai Sea oilfield; they were found to degrade marine oil spills effectively. The experimental results show that the degradation efficiency of crude oil with these four strains （XT-4, SZ-1-25, B-4-9, BS-3-12） is 95.97%, 96.01%, 97.99% and 98.99%, respectively, in their optimum conditions. The characteristics of bacterial biodegradation are investigated. The simulation biotreatment of oil-contaminated beach sand, with an initial oil content of 5,664mg/kg-dry-sand, shows that the residual oil content is 2,700 mg/kg-dry-sand and 2,679 mg/kg-dry-sand after 170 days＇ treatment with two bacteria （B-4-9, BS-3- 12）, respectively.

Biogeographical note on Antarctic microforae: Endemism and cosmopolitanism

This study deals with the biogeography of Antarctic microflora （Antarctica acts as best model to study microbial biogeography） such as cyanobacteria and selected halophiles with special emphasis on Halomonas variabilis and Bacillus licheniformis.Halophiles are known to be resistant not only to salt stress,but also to extreme temperature,pressure,and aridity and they are capable of surviving in harsh environments such as polar regions,deep-sea habitats,and deserts.Many microbes are known to be resistant to hostile environmental conditions,and are capable of surviving in harsh environments.Our group has isolated 444 strains belonging to 28 genera of halophiles from various environments around the world.The 16S rRNA gene sequences revealed that many of the isolated strains from geographically distant habitats having different environmental conditions,were closely related to each other,with some strains possessing 100％ identical sequences.Organisms possessing survival mechanism such as spore formation are usually ubiquitous.The genus Halomonas is represented by potentially endemic strains and the ubiquitous H.variabilis,while spore-forming B.licheniformis showed cosmopolitan distribution.One potentially endemic （moderate endemicity that is regional and/or continental distribution） strain was reported from Syowa station,East Antarctica,and Mario Zucchelli station,West Antarctica,which are geographically separated by 3000 km.Moreover,15 strains having 100％ similarity with B.licheniformis were considered cosmopolitans.The results of this work provide support for the middle-ground model that some microbes have moderate endemicity and others have cosmopolitan distribution.These results will contribute to a greater understanding of microbial biogeography with special emphasis on Antarctica.

Effects of Sea Salts on Induction of Cell Proliferation in Liquid Cultures of Mangrove Plants, Sonneratia caseolaris and S. alba

The effects of five salt ingredients of sea water, KCl, NaCl, CaCl2, MgCl2 and MgSO4, on induction of cell prolif-eration in Sonneratia caseolaris were investigated. Proliferation was examined in tissue explants derived from such as leaves, cotyledons, and hypocotyls using a small-scale liquid culture method. Addition of 12.5-25 mM of MgCl2 was unique in stimulating cell proliferation in all tissues of S. caseolaris. Otherwise, different effects of salts were observed among the three tissues. In hypocotyl culture, 25-50 mM of NaCl and CaCl2 stimulated cell divisions. Tolerance to 100 mM of MgSO4 was observed in leaves. Three osmotically active compounds commonly used in tissue culture, sorbitol, mannitol and glycinebetaine, were also tested to assess the importance of osmotic effects on cell proliferation. No significant stimulation by these was observed over a wide range of concentrations. Data were compared with those of cotyledon cultures of another mangrove, S. alba, which exhibits no stimulation by MgCl2, stimulation by KCl and tolerance to NaCl. Mechanisms for adaptation of mangrove plants to various and high salts were discussed by comparing the differences in reaction to salts in cultures of two Sonneratia mangrove species of the same genera growing different salt environment.

Salt tolerant culturable microbes accessible in the soil of the Sundarban Mangrove forest, India

Sundarban Mangrove forest is highly productive marine ecosystem where halophilic microbes actively participate in bio-mineralization and biotransformation of minerals. The population of aerobic halophilic microbes was studied to determine their distribution with the availability of different physicochemical parameters with increasing depth of this forest sediment. The present study revealed that microbes present in the top soil region were less tolerant to fluctuation in salinity than the middle and bottom segment. Microbes isolated from bottom segment showed higher growth rate in anaerobic condition. A decreasing trend of total microbial population and organic carbon content of soil were found with increase in depth. In contrary a reverse profile was found for salinity. A significant stratification was found to exist among microbial population and the salty nature of the soil of Sundarban Mangrove forest.

Germination Behaviour of a Perennial Amaranthaceae of North Western Algeria(Oran):Arthrocnemum glaucum(Del.)Ung.Respectively Submitted to Thermal Factor and Salt Stress

In light of the importance of seeds germinative phase in the progress of later stages of development of any particular plant species in semi-arid and arid area, it is essential to study the germinative behavior and to evaluate the tolerance of a perennial halophyte of Amaranthacaea family, Arthrocnemum glaucum (Del.) Ung. during this crucial phase. A study is devoted to this species seeds germination, which although it’s a mandatory halophyte, it’s less tolerant to salt stress in germination’ phase. Germinative tests are carried out in laboratory, on seeds putted in Petri dishes placed at different temperatures. They enable us to determine the optimal germination temperature, which is 40&#176C while the cold one (5&#176C) completely inhibits it. The seeds of Arthrocnemum glaucum show an optimum germination of 44% in distilled water (NaCl 0 mm) similar to glycophytes but they differ in their ability to germinate at higher salinities. Beyond 300 mM germination is completely inhibited. Prior to break dormancy seeds, a cold treatment at 5&#176C has been achieved;the germination capacity of the seeds increases with the duration of the cold treatment. It increases from 30% after 2 weeks of treatment to 80% after a treatment of 27 weeks.

Characterization and recovery of the halophiles in the fluid inclusions in halite

1 Introduction It is the focus of geology and biology that the creature preserved in the geological history and the organic evolution.The creature preserved in geological history by these things:sedimentary,frozen earth,chrysophoron and evaporation salt.Evaporation salt can preserve the microbe

Isolation and Identity of Halophiles in The Floating Bath Water in Yuncheng

1 Introduction The study of floating bath began in 1914,by Dor.John C.Lily,the expert of medical and neurology.She has done different kinds of floating experiments to study about the origin of consciousness and the relationship with brain.Previous research showed that the effect of floating

Ancient Microorganisms and Carotenoids Preserved in Fluid Inclusions in Halite from Chaka Salt Lake,Western China:Evidence from Micro-observation and in situ Raman Spectroscopy

Trapped ancient microorganisms in halite fluid inclusions are of special interest to the understanding of biology and ecology in salt lake systems.With the integration of petrologic,microthermometric,and Raman spectroscopic analyses,this study utilizes fluid inclusions from Chaka Salt Lake,eastern Qaidam Basin,NW China,to assess the possibility of microorganism-trapping by fluid inclusions.Here,we report that the solid phase of some primary fluid inclusions contains carotenoids,which is interpreted as evidence of Dunaliella algae,and that the coexisting liquid phase comprises SO_(4)^(2-).The homogenization temperatures of single-phase primary fluid inclusions indicate that the precipitation temperature of the Holocene halite in Chaka Salt Lake ranges from 13.5℃ to 36.4℃.This suggests that fluid inclusions in halite are a good medium for trapping and preserving ancient microorganisms and organic matter in salt lakes,and that Raman spectroscopy has good potential to identify halophilic archaea.

THE DISCOVERY AND ITS GEOLOGICAL SIGNIFICANCE OF FOSSIL FAECAL PELLETS OF BRINE SHRIMPS IN MIRABILITE DEPOSIT

The brine shrimp, Artemia salina, is affiliated with a lineage of Arthropoda-class Crustacea-subclass Branchiopodaorder Anostraca-family Arlemiidae. The individuals, 10 to 11 mm long for male but slightly longer for female, show a striking adaptation to high-salinity waters and salt pans, for example, in Great Salt Lake of the U. S. A., a num-