Biomedical subjects

Ken Takai

Publications and source records attributed to Ken Takai.

49 records · Page 3Linked to original sources

PubMed2003

Related assemblages of sulphate-reducing bacteria associated with ultradeep gold mines of South Africa and deep basalt aquifers of Washington State.

We characterized the diversity of sulphate-reducing bacteria (SRB) associated with South African gold mine boreholes and deep aquifer systems in Washington State, USA. Sterile cartridges filled with crushed country rock were installed on two hydrologically isolated and chemically distinct sites at depths of 3.2 and 2.7 km below the land surface (kmbls) to allow development of biofilms. Enrichments of sulphate-reducing chemolithotrophic (H2) and organotrophic (lactate) bacteria were established from each site under both meso- and thermophilic conditions. Dissimilatory sulphite reductase (Dsr) and 16S ribosomal RNA (rRNA) genes amplified from DNA extracted from the cartridges were most closely related to the Gram-positive species Desulfotomaculum thermosapovorans and Desulfotomaculum geothermicum, or affiliated with a novel deeply branching clade. The dsr sequences recovered from the Washington State deep aquifer systems affiliated closely with the South African sequences, suggesting that Gram-positive sulphate-reducing bacteria are widely distributed in the deep subsurface.

Biofilms↗

PubMed2003

Shifts in archaeal communities associated with lithological and geochemical variations in subsurface Cretaceous rock.

Subsurface microbial community structure in relation to geochemical gradients and lithology was investigated using a combination of molecular phylogenetic and geochemical analyses. Discreet groundwater and substratum samples were obtained from depths ranging from 182 to 190 m beneath the surface at approximately 10-cm intervals using a multilevel sampler (MLS) that straddled Cretaceous shale and sandstone formations at a site in the southern San Juan Basin in New Mexico. DNA and RNA were extracted directly from quartzite sand substratum loaded into individual cells of the MLS and colonized in situ. Polymerase chain reaction (PCR)-mediated T-RFLP analysis of archaeal rRNA genes (rDNA) in conjunction with partial sequencing analysis of archaeal rDNA libraries and quantitative RNA hybridization with oligonucleotide probes were used to probe community structure and function. Although total microbial populations remained relatively constant over the entire depth interval sampled, significant shifts in archaeal populations, predominantly methanogens, were observed. These shifts coincided with the geochemical transition from relatively high methane (26 mM), low sulphate (< 3 mg l(-1)) conditions in the region adjacent to the organic matter-rich shale to relatively low-methane (< 0.5 mM), high-sulphate (48 mg l(-1)) conditions in the organic-poor sandstone beneath the shale. These results indicated that active, phylogenetically diverse archaeal communities were present in the subsurface Cretaceous rock environment at this site and that major archaeal clades shifted dramatically over scales of tens of centimetres, corresponding to changes in the lithology and geochemical gradients.

Archaea↗

PubMed2003

Marinithermus hydrothermalis gen. nov., sp. nov., a strictly aerobic, thermophilic bacterium from a deep-sea hydrothermal vent chimney.

A novel thermophilic marine bacterium, designated strain T1T, was isolated from a deep-sea hydrothermal vent chimney sample collected from the Suiyo Seamount in the Izu-Bonin Arc, Japan, at a depth of 1,385 m. Cells of strain T1T were rod-shaped, occurring in pairs or filamentous, and stained Gram-negative. Growth was observed between 50.0 and 72.5 degrees C (optimum 67.5 degrees C; 30 min doubling time) and at pH 6.25-7.75 (optimum pH 7.00). The isolate absolutely required NaCl, at a concentration of 0.5-4.5% (optimum 30%). It was a strictly aerobic heterotroph capable of growing solely on complex organic substrates such as yeast extract, tryptone and Casamino acids, utilizing glutamate, proline, serine, cellobiose, trehalose, sucrose, acetate and pyruvate as complementary substrates. The G+C content of the genomic DNA was 68.6 mol%. The 16S rRNA gene sequence of the isolate was most similar to those from members of the genus Thermus, but the isolate was distantly related to them at the genus level (<90%). In addition, phylogenetic analysis indicated that the isolate was on a novel lineage, deeply branched prior to divergence of the genus Thermus. On the basis of phylogenetic analysis and physiological traits of the isolate, it should be described as a member of a novel genus distinct from the previously described genus Thermus. The name Marinithermus gen. nov. is proposed, with Marinithermus hydrothermalis gen. nov., sp. nov. as the type species. The type strain of M. hydrothermalis gen. nov., sp. nov. is strain T1T (=JCM 11576T =DSM 14884T).

Base Composition↗

PubMed2003

Deferribacter desulfuricans sp. nov., a novel sulfur-, nitrate- and arsenate-reducing thermophile isolated from a deep-sea hydrothermal vent.

A novel anaerobic, heterotrophic thermophile was isolated from a deep-sea hydrothermal vent chimney at the Suiyo Seamount in the Izu-Bonin Arc, Japan. The cells were bent, flexible rods, with a single polar flagellum. Growth was observed between 40 and 70 degrees C (optimum temperature: 60-65 degrees C; doubling time, 40 min) and between pH 5.0 and 7.5(optimum pH 6.5). The isolate was a strictly anaerobic heterotroph capable of using complex organic compounds (yeast extract, tryptone, peptone, casein and Casamino acids), ethanol and various organic acids as energy and carbon sources. Hydrogen could serve as a supplementary energy source. Elemental sulfur (S(0)), nitrate or arsenate was required for growth as an electron acceptor. The G + C content of the genomic DNA was 38.6 mol%. Phylogenetic analysis based on 16S rDNA sequences indicated that isolate SSM1(T) is closely related to Deferribacter thermophilus BMA(T) (98.1%). However, the novel isolate could be clearly differentiated from D. thermophilus BMA(T) on the basis of its physiological and genetic properties. The name Deferribacter desulfuricans sp. nov. (type strain SSM1(T) = JCM 11476(T) = DSM 14783(T)) is proposed.

Arsenates↗

PubMed2003

Persephonella hydrogeniphila sp. nov., a novel thermophilic, hydrogen-oxidizing bacterium from a deep-sea hydrothermal vent chimney.

A novel thermophilic, hydrogen-oxidizing bacterium, designated 29W(T), was isolated from a deep-sea hydrothermal vent chimney sample collected from the Suiyo Seamount in the Izu-Bonin Arc, Japan, at a depth of 1385 m. The cells were cocci (0.9-1.0 microm in diameter) and straight rods (2.3-2.7 microm long) under static and agitated culture conditions, respectively. The new isolate was an obligate chemolithoautotroph growing by respiratory nitrate reduction with H2, forming N2 as a final product. A very low concentration of O2 (optimum 0.6-0.8%, v/v) was also used as an alternative electron acceptor while reduced sulfur compounds did not serve as electron donors. Anoxic hydrogen-oxidizing growth with nitrate was observed between 50 and 72.5 degrees C (optimum 70 degrees C; 40 min doubling time), pH 5.5 and 7.6 (optimum pH 7.2), and in the presence of 1.5 and 5.0% NaCl (optimum 2.5%). The G + C content of the genomic DNA was 37.3 mol%. Phylogenetic analysis based on 16S rDNA sequences indicated that the isolate was a member of the recently described genus Persephonella in a potential new family within the order Aquificales. On the basis of the physiological and molecular properties of the new isolate, the name Persephonella hydrogeniphila sp. nov. is proposed. The type strain is strain 29W(T) (= JCM 11663(T) = DSM 15103(T)).

Bacterial Typing Techniques↗

PubMed2003

Sulfurihydrogenibium subterraneum gen. nov., sp. nov., from a subsurface hot aquifer.

A polyphasic taxonomic study was performed on a novel facultatively anaerobic, hydrogen- or sulfur/thiosulfate-oxidizing, thermophilic chemolithoautotroph recently isolated from subsurface hot aquifer water in a Japanese gold mine. The cells were straight to slightly curved rods, with a single polar flagellum. Growth was observed at 40-70 degrees C (optimum 60-65 degrees C; 80 min doubling time) and at pH 6.4-8.8 (optimum pH 7.5). The isolate was unable to use complex organic compounds, carbohydrates, amino acids or organic acids as sole energy and carbon sources. The G + C content of the genomic DNA was 31.3 mol%. Phylogenetic analysis based on 16S rDNA sequences indicated that the isolate was closely related to an uncultivated group of micro-organisms within the order Aquificales obtained from Icelandic and Japanese hot spring microbial mats, but distantly related to previously identified genera of the Aquificales such as Persephonella, Aquifex and Hydrogenobacter. The name Sulfurihydrogenibium subterraneum gen. nov., sp. nov. is proposed for this novel species; the type strain is HGMK1(T) (= JCM 11477(T) = ATCC BAA-562(T) = DSM 15120(T)).

Bacterial Typing Techniques↗

PubMed2003

Sulfurimonas autotrophica gen. nov., sp. nov., a novel sulfur-oxidizing epsilon-proteobacterium isolated from hydrothermal sediments in the Mid-Okinawa Trough.

A novel mesophilic, sulfur- and thiosulfate-oxidizing bacterium, strain OK10(T), was isolated from deep-sea sediments at the Hatoma Knoll in the Mid-Okinawa Trough hydrothermal field. Cells of strain OK10(T) were short rods, each being motile by means of a single polar flagellum. The isolate grew at 10-40 degrees C (optimum 25 degrees C) and pH 4.5-9.0 (optimum pH 6.5). It grew chemolithoautotrophically with elemental sulfur, sulfide and thiosulfate as sole electron donors and oxygen as electron acceptor. Molecular hydrogen did not support growth. The G+C content of the genomic DNA of strain OK10(T) was 35.2 mol%. Phylogenetic analysis, based on 16S rRNA gene sequences, indicated that the isolate belonged to the epsilon-Proteobacteria. On the basis of its physiological and molecular characteristics, strain OK10(T) (=ATCC BAA-671(T)=JCM 11897(T)) represents the sole species of a new genus, Sulfurimonas, for which the name Sulfurimonas autotrophica is proposed.

Campylobacter↗

PubMed2003

Balnearium lithotrophicum gen. nov., sp. nov., a novel thermophilic, strictly anaerobic, hydrogen-oxidizing chemolithoautotroph isolated from a black smoker chimney in the Suiyo Seamount hydrothermal system.

A novel, extremely thermophilic bacterium, designated strain 17S(T), was isolated from a deep-sea hydrothermal vent chimney at the Suiyo Seamount in the Izu-Bonin Arc, Japan. The cells were rods with no apparent motility, most of which were narrow in the middle in the exponential-growth phase and had several polar flagella at both ends. Growth was observed between 45 and 80 degrees C (optimum temperature, 70-75 degrees C; doubling time, 80 min) and between pH 5.0 and 7.0 (optimum pH, 5.4). The isolate was a strictly anaerobic chemolithoautotroph that was capable of using molecular hydrogen as its sole energy source and carbon dioxide as its sole carbon source. Elemental sulfur (S(0)) was required for growth as an electron acceptor. The G+C content of the genomic DNA was 34.6 mol%. Phylogenetic analysis based on 16S rDNA sequences indicated that the isolate was related to Thermovibrio ruber ED11/3LLK(T) and Desulfurobacterium thermolithotrophum BSA(T), whilst it appeared to be a novel lineage prior to the divergence of these genera. This isolate could also be differentiated from both T. ruber ED11/3LLK(T) and D. thermolithotrophum BSA(T) on the basis of physiological properties. The name Balnearium lithotrophicum gen. nov., sp. nov. is proposed for this isolate (type strain, 17S(T)=JCM 11970(T)=ATCC BAA-736(T)).

Bacteria↗

PubMed2003

Microbial communities associated with geological horizons in coastal subseafloor sediments from the sea of okhotsk.

Microbial communities from a subseafloor sediment core from the southwestern Sea of Okhotsk were evaluated by performing both cultivation-dependent and cultivation-independent (molecular) analyses. The core, which extended 58.1 m below the seafloor, was composed of pelagic clays with several volcanic ash layers containing fine pumice grains. Direct cell counting and quantitative PCR analysis of archaeal and bacterial 16S rRNA gene fragments indicated that the bacterial populations in the ash layers were approximately 2 to 10 times larger than those in the clays. Partial sequences of 1,210 rRNA gene clones revealed that there were qualitative differences in the microbial communities from the two different types of layers. Two phylogenetically distinct archaeal assemblages in the Crenarchaeota, the miscellaneous crenarchaeotic group and the deep-sea archaeal group, were the most predominant archaeal 16S rRNA gene components in the ash layers and the pelagic clays, respectively. Clones of 16S rRNA gene sequences from members of the gamma subclass of the class Proteobacteria dominated the ash layers, whereas sequences from members of the candidate division OP9 and the green nonsulfur bacteria dominated the pelagic clay environments. Molecular (16S rRNA gene sequence) analysis of 181 isolated colonies revealed that there was regional proliferation of viable heterotrophic mesophiles in the volcanic ash layers, along with some gram-positive bacteria and actinobacteria. The porous ash layers, which ranged in age from tens of thousands of years to hundreds of thousands of years, thus appear to be discrete microbial habitats within the coastal subseafloor clay sediment, which are capable of harboring microbial communities that are very distinct from the communities in the more abundant pelagic clays.

Colony Count, Microbial↗

PubMed2002

Thermaerobacter nagasakiensis sp. nov., a novel aerobic and extremely thermophilic marine bacterium.

A novel, extremely thermophilic bacterium was isolated from a shallow marine hydrothermal vent at depth of 22 m in Tachibana Bay, Nagasaki Prefecture, Japan. Cells were gram-negative, non-spore-forming, motile rods. Growth was observed between 52 and 78 degrees C (optimum 70 degrees C), pH 5 and 8 (optimum pH 7) and 0-4.5% NaCl (optimum 1.0%). The isolate was a strictly aerobic heterotroph utilizing yeast extract and trypticase peptone. The G+C content of the genomic DNA is 69 mol%. Analysis of 16S rDNA sequences indicated that strain Ts1a is closely related to Thermaerobacter marianensis. The differences in physiology and DNA-DNA similarity between strain Ts1a and T. marianensis showed that strain Ts1a represents a new species of Thermaerobacter. The type strain of T. nagasakiensis is strain Ts1a (=JCM11223, DSM 14512).

Bacillaceae↗

PubMed2002

Methanothermococcus okinawensis sp. nov., a thermophilic, methane-producing archaeon isolated from a Western Pacific deep-sea hydrothermal vent system.

A novel thermophilic, methane-producing archaeon was isolated from a deep-sea hydrothermal vent chimney at the Iheya Ridge, in the Okinawa Trough, Japan. The cells were highly motile, irregular cocci, with a polar bundle of flagella. Growth was observed between 40 and 75 degrees C (optimum 60-65 degrees C; 30 min doubling time) and between pH 4.5 and 8.5 (optimum pH 6.7). The isolate was a strictly anaerobic autotroph capable of using hydrogen and carbon dioxide as sole sources of energy and carbon. Formate can serve as an alternative energy source. The G+C content of the genomic DNA was 33.5 mol%. Phylogenetic analysis based on 16S rDNA sequences and DNA-DNA hybridization analysis indicated that the isolate was closely related to members of the genera Methanococcus and Methanothermococcus. This isolate, however, could be differentiated from the previously described species of these genera on the basis of its physiological and molecular properties. The name Methanothermococcus okinawensis sp. nov is proposed, with the type strain IH1T (=JCM 11175T=DSM 14208T).

DNA, Ribosomal↗

PubMed2002

Isolation and metabolic characteristics of previously uncultured members of the order aquificales in a subsurface gold mine.

Culture-dependent and -independent techniques were combined to characterize the physiological properties and the ecological impacts of culture-resistant phylotypes of thermophiles within the order Aquificales from a subsurface hot aquifer of a Japanese gold mine. Thermophilic bacteria phylogenetically associated with previously uncultured phylotypes of Aquificales were successfully isolated. 16S ribosomal DNA clone analysis of the entire microbial DNA assemblage and fluorescence in situ whole-cell hybridization analysis indicated that the isolates dominated the microbial population in the subsurface aquifer. The isolates were facultatively anaerobic, hydrogen- or sulfur/thiosulfate-oxidizing, thermophilic chemolithoautotrophs utilizing molecular oxygen, nitrate, ferric iron, arsenate, selenate, and selenite as electron acceptors. Their versatile energy-generating systems may reflect the geochemical conditions of their habitat in the geothermally active subsurface gold mine.

Bacteria↗

PubMed1998

ppc, the gene for phosphoenolpyruvate carboxylase from an extremely thermophilic bacterium, Rhodothermus obamensis: cloning, sequencing and overexpression in Escherichia coli.

The ppc gene, which encodes phosphoenolpyruvate carboxylase (PEPC) of an extremely thermophilic bacterium, Rhodothermus obamensis, was directly sequenced by the thermal asymmetric interlaced (TAIL) PCR method. An ORF for a 937 amino acid polypeptide was found in the gene. The ppc gene had a high G+C content (66.2 mol%) and the third position of the codon exhibited strong preference for G or C usage (85.0 mol%). The calculated molecular mass was 107,848 Da, which was consistent with the molecular mass of the enzyme as determined by SDS-PAGE (100 kDa). The amino acid sequence of R. obamensis PEPC was closely related to that of PEPC from another thermophile, a Thermus sp., and from a mesophile, Corynebacterium glutamicum, exhibiting 45.3% or 37.7% identity and 61.5% or 56.5% similarity, respectively. By Southern analysis, the ppc gene was found to be present in a single copy in the genomic DNA of this organism. The cloned gene was expressed in Escherichia coli using a pET expression vector system and a thermostable recombinant PEPC was obtained. Comparison of the deduced amino acid sequences of the thermophilic and mesophilic PEPCs revealed distinct or common preferences for specific amino acid composition and substitutions in the two thermophilic enzymes.

Amino Acid Sequence↗