| atp-independent dna topoisomerase from fervidobacterium islandicum. | thermotogales are thermophilic eubacteria belonging to a very slowly evolving branch in the eubacterial tree. in this report, we describe the purification and characterization of an atp-independent dna topoisomerase from the thermotogale, fervidobacterium islandicum. the enzyme, a monomer of about 75 kda, is a type i dna topoisomerase sharing many properties with the other bacterial topoisomerases i: it absolutely requires mg2+ for activity, relaxes negatively but not positively supercoiled dna ... | 1993 | 8241262 |
| fervidobacterium gondwanense sp. nov., a new thermophilic anaerobic bacterium isolated from nonvolcanically heated geothermal waters of the great artesian basin of australia. | a new thermophilic, carbohydrate-fermenting, obligately anaerobic bacterial species was isolated from a runoff channel formed from flowing bore water from the geothermally heated aquifer of the great artesian basin of australia. the cells of this organism were nonsporulating, motile, gram negative, and rod shaped and generally occurred singly or in pairs. the optimum temperature for growth was 65 to 68 degrees c, and no growth occurred at temperatures below 44 degrees c or above 80 degrees c. gr ... | 1996 | 8573506 |
| new compatible solutes related to di-myo-inositol-phosphate in members of the order thermotogales. | the accumulation of intracellular organic solutes was examined in six species of the order thermotogales by nuclear magnetic resonance spectroscopy. the newly discovered compounds di-2-o-beta-mannosyl-di-myo-inositol-1,1'(3,3')-phosphate and di-myo-inositol-1,3'-phosphate were identified in thermotoga maritima and thermotoga neapolitana. in the latter species, at the optimum temperature and salinity the organic solute pool was composed of di-myo-inositol-1,1'(3,3')-phosphate, beta-glutamate, and ... | 1996 | 8824608 |
| estimation of genome sizes of hyperthermophiles. | genomes of various hyperthermophilic and extremely thermophilic prokaryotes were analyzed with respect to size, physical organization, and 16s rdna copy number. our results show that all the genomes are circular, and they are in the size range of 1.6-1.8 mb for pyrodictium abyssi, methanococcus igneus, pyrobaculum aerophilum, archaeoglobus fulgidus, archaeoglobus lithotrophicus, and archaeoglobus profundus (the two bacteria fervidobacterium islandicum and thermosipho africanus possess genomes of ... | 1998 | 9672684 |
| pullulanase type i from fervidobacterium pennavorans ven5: cloning, sequencing, and expression of the gene and biochemical characterization of the recombinant enzyme. | the gene encoding the type i pullulanase from the extremely thermophilic anaerobic bacterium fervidobacterium pennavorans ven5 was cloned and sequenced in escherichia coli. the pula gene from f. pennavorans ven5 had 50.1% pairwise amino acid identity with pula from the anaerobic hyperthermophile thermotoga maritima and contained the four regions conserved among all amylolytic enzymes. the pullulanase gene (pula) encodes a protein of 849 amino acids with a 28-residue signal peptide. the pula gene ... | 1999 | 10224005 |
| thiosulfate reduction and alanine production in glucose fermentation by members of the genus coprothermobacter. | coprothermobacter platensis is an anaerobic, proteolytic, thermophilic bacterium, which is phylogenetically related to the genera fervidobacterium and thermotoga. the organism was found to reduce thiosulfate to sulfide during growth on carbohydrates and proteinaceous substrates. growth on glucose was inhibited by hydrogen, but this inhibition was overcome by thiosulfate reduction, stirring, increasing the headspace volume and coculturing with a hydrogen-consuming methanogen. alanine was detected ... | 2000 | 10959561 |
| crystallization and preliminary x-ray crystallographic studies of the thermoactive pullulanase type i, hydrolyzing alpha-1,6 glycosidic linkages, from fervidobacterium pennivorans ven5. | crystals of the thermoactive recombinant f. pennivorans type i pullulanase, purified from the supernatant of a bacillus subtilis culture, have been obtained by the vapour-diffusion method in the presence of the inhibitor beta-cyclodextrin (2 mm) by mixing protein (15 mg ml(-1)) with an equal volume of crystallization solution containing 0.1 m bis-tris propane ph 6.5, 50 mm mgcl(2) and 15% polyethylene glycol 3350. crystals diffracted to 3.0 a using conventional cu kalpha radiation and belong to ... | 2000 | 11053856 |
| polyamines of the thermophilic eubacteria belonging to the genera thermosipho, thermaerobacter and caldicellulosiruptor. | cellular polyamines of four new thermophiles located in three early branched eubacterial clades, were investigated for the chemotaxonomic significance of polyamine distribution profiles. the thermophilic anaerobic thermosipho japonicus, belonging to the order thermotogales, contained norspermidine, norspermine and thermospermine in addition to spermidine and spermine. the polyamine profile was identical to the polyamine composition of thermotoga, fervidobacterium and petrotoga species of the ord ... | 2001 | 11327112 |
| molecular characterization of fervidolysin, a subtilisin-like serine protease from the thermophilic bacterium fervidobacterium pennivorans. | the fls gene encoding fervidolysin, a keratin-degrading proteolytic enzyme from the thermophilic bacterium fervidobacterium pennivorans, was isolated using degenerate primers combined with southern hybridization and inverse polymerase chain reaction. further sequence characterization demonstrated that the 2.1-kb fls gene encoded a 699-amino-acid preproenzyme showing high homology with the subtilisin family of the serine proteases. it was cloned into a pet9d vector, without its signal sequence, a ... | 2002 | 12072953 |
| development of fluorescent adjacent hybridization probes and their application in real-time pcr for the simultaneous detection and identification of fervidobacterium and caloramator. | a stretch of nucleotides consisting of a conserved region and an adjacent upstream variable region in the 16s rdna of members of domain bacteria was identified as a suitable target site for developing a real-time pcr adjacent hybridization assay. a single universal fluorogenic cyanin 5-labelled probe, cy5 1046+, targeting the conserved region, and two fitc-labelled probes, calo and fervi, targeting the variable region were designed and synthesized for the identification and differentiation of th ... | 2002 | 12361295 |
| native-feather degradation by fervidobacterium islandicum aw-1, a newly isolated keratinase-producing thermophilic anaerobe. | a native-feather-degrading thermophilic anaerobe was isolated from a geothermal hot stream in indonesia. isolate aw-1, identified as a member of the species fervidobacterium islandicum, was shown to degrade native feathers (0.8%, w/v) completely at 70 degrees c and ph 7 with a maximum specific growth rate (0.14 h(-1)) in thermotoga- fervidobacterium(tf) medium. after 24 h of culture, feather degradation led to an increase in free amino acids such as histidine, cysteine and lysine. moreover, nutr ... | 2002 | 12420177 |
| crystal structure of fervidolysin from fervidobacterium pennivorans, a keratinolytic enzyme related to subtilisin. | structure-forming fibrous proteins like keratins, gelatins and collagens are degraded only by a few proteases as their tight packing limits access to the potential cleavage sites. to understand the keratin degradation in detail, we describe the first crystal structure of a keratin-degrading enzyme (keratinase), fervidolysin, from fervidobacterium pennivorans as an immature form with propeptide (pd)-bound. the 1.7a resolution crystal structure shows that the protease is composed of four domains: ... | 2004 | 14687574 |
| cloning, sequencing, and characterization of a heat- and alkali-stable type i pullulanase from anaerobranca gottschalkii. | the gene encoding a type i pullulanase was identified from the genome sequence of the anaerobic thermoalkaliphilic bacterium anaerobranca gottschalkii. in addition, the homologous gene was isolated from a gene library of anaerobranca horikoshii and sequenced. the proteins encoded by these two genes showed 39% amino acid sequence identity to the pullulanases from the thermophilic anaerobic bacteria fervidobacterium pennivorans and thermotoga maritima. the pullulanase gene from a. gottschalkii (en ... | 2004 | 15184138 |
| h(2) production and carbon utilization by thermotoga neapolitana under anaerobic and microaerobic growth conditions. | h(2) production by petrotoga miotherma, thermosipho africanus, thermotoga elfii, fervidobacterium pennavorans, and thermotoga neapolitana was compared under microaerobic conditions. contrary to these previously reported strains being strict anaerobes, all tested strains grew and produced h(2) in the presence of micromolar levels of o(2). t. neapolitana showed the highest h(2) production under these conditions. microscopic counting techniques were used to determine growth curves and doubling time ... | 2004 | 15289678 |
| cloning and expression of islandisin, a new thermostable subtilisin from fervidobacterium islandicum, in escherichia coli. | a gene encoding a subtilisin-like protease, designated islandisin, from the extremely thermophilic bacterium fervidobacterium islandicum (dsmz 5733) was cloned and actively expressed in escherichia coli. the gene was identified by pcr using degenerated primers based on conserved regions around two of the three catalytic residues (asp, his, and ser) of subtilisin-like serine protease-encoding genes. using inverse pcr regions flanking the catalytic residues, the gene could be cloned. sequencing re ... | 2005 | 16000809 |
| thiosulfate reduction, an important physiological feature shared by members of the order thermotogales. | several members of the order thermotogales in the domain bacteria, viz., thermotoga neapolitana, thermotoga maritima, thermosipho africanus, fervidobacterium islandicum, and thermotoga strain sebr 2665, an isolate from an oil well, reduced thiosulfate to sulfide. this reductive process enhanced cellular yields and growth rates of all the members but was more significant with the two hyperthermophiles t. neapolitana and t. maritima. this is the first report of such an occurrence in this group of ... | 1995 | 16535035 |
| keratin degradation by fervidobacterium pennavorans, a novel thermophilic anaerobic species of the order thermotogales. | from a hot spring of the azores islands a novel thermophilic bacterium belonging to the thermotogales order was isolated. this strain, which grows optimally at 70(deg)c and ph 6.5, is the first known extreme thermophile that is able to degrade native feathers at high temperatures. the enzyme system converts feather meal to amino acids and peptides. on the basis of physiological, morphological, and 16s rdna studies the new isolate was found to be a member of the thermotogales order and was identi ... | 1996 | 16535379 |
| purification and properties of a thermostable pullulanase from a newly isolated thermophilic anaerobic bacterium, fervidobacterium pennavorans ven5. | extremely thermophilic anaerobic fermentative bacteria growing at temperatures between 50 and 80(deg)c (optimum, 65 to 70(deg)c) were isolated from mud samples collected at abano terme spa (italy). the cells were gram-negative motile rods, about 1.8 (mu)m in length and 0.6 (mu)m in width, occurring singly and in pairs. cells commonly formed spheroids at one end similar to fervidobacterium islandicum and fervidobacterium nodosum. the new isolate differs from f. nodosum by the 7% higher g+c conten ... | 1997 | 16535541 |
| fgoi, a type ii restriction endonuclease from the thermoanaerobe fervidobacterium gondwanense ab39(t). | restriction endonuclease activity was detected in 11 out of 13 fervidobacterium isolates, including f. islandicum h21(t), f. gondwanense ab39(t), and nine other fervidobacterium-like strains isolated from the great artesian basin of australia. the restriction endonuclease from f. gondwanense ab39(t) was partially purified and designated fgoi. fgoi recognized a 4 nucleotide sequence 5'-ctag-3' and cleaved between nucleotides c and t to produce a 2 base 5' overhang (5'-c/tag-3'). as predicted from ... | 1998 | 16887647 |
| structural characterization of diabolic acid-based tetraester, tetraether and mixed ether/ester, membrane-spanning lipids of bacteria from the order thermotogales. | the distribution of core lipids in the membranes of nine different species of the order thermotogales, one of the early and deep branching lineages in the bacteria, were examined by hplc/ms and demonstrated to consist of membrane-spanning diglycerol lipids comprised of diabolic acid-derived alkyl moieties. in the thermotoga species the core membrane lipids are characterized by the presence of both ester and ether bonds, whereas in the phylogenetically more distinct thermosipho and fervidobacteri ... | 2007 | 17643227 |
| fervidobacterium changbaicum sp. nov., a novel thermophilic anaerobic bacterium isolated from a hot spring of the changbai mountains, china. | a thermophilic, obligately anaerobic, rod-shaped bacterium (strain cbs-1(t)) was isolated from a hot spring mixture of water and mud of the changbai mountains, china. strain cbs-1(t) was found to be non-sporulating, gram-negative, with optimal growth at 75-80 degrees c. it grew on a wide range of carbon sources, including glucose, lactose, maltose, starch, sorbitol and pyruvate amongst others. the dna g+c content of strain cbs-1(t) was 31.9 mol%. the 16s rrna gene sequence analysis indicated tha ... | 2007 | 17911306 |
| evolution of mal abc transporter operons in the thermococcales and thermotogales. | the mal genes that encode maltose transporters have undergone extensive lateral transfer among ancestors of the archaea thermococcus litoralis and pyrococcus furiosus. bacterial hyperthermophiles of the order thermotogales live among these archaea and so may have shared in these transfers. the genome sequence of thermotoga maritima bears evidence of extensive acquisition of archaeal genes, so its ancestors clearly had the capacity to do so. we examined deep phylogenetic relationships among the m ... | 2008 | 18197971 |
| evaluation of gfp tag as a screening reporter in directed evolution of a hyperthermophilic beta-glucosidase. | by applying a directed evolution methodology specific enzymatic characteristics can be enhanced, but to select mutants of interest from a large mutant bank, this approach requires high throughput screening and facile selection. to facilitate such primary screening of enhanced clones, an expression system was tested that uses a green fluorescent protein (gfp) tag from aequorea victoria linked to the enzyme of interest. as gfp's fluorescence is readily measured, and as there is a 1:1 molar correla ... | 2009 | 19214809 |
| crystallization and preliminary crystallographic analysis of thermophilic cellulase from fervidobacterium nodosum rt17-b1. | fncel5a, a thermostable endoglucanase, is a member of glycohydrolase family 5 which catalyzes the hydrolysis of cellulose to glucose in the thermophilic bacterium fervidobacterium nodosum rt17-b1. fncel5a is particularly interesting because of its high thermostability (t(opt) = 353 k, half-life 48 h) and its high specific activity towards carboxymethylcellulose. these properties make fncel5a an attractive target for protein engineering to improve cellulase activity. in order to resolve the cryst ... | 2009 | 19255469 |
| on the chimeric nature, thermophilic origin, and phylogenetic placement of the thermotogales. | since publication of the first thermotogales genome, thermotoga maritima strain msb8, single- and multi-gene analyses have disagreed on the phylogenetic position of this order of bacteria. here we present the genome sequences of 4 additional members of the thermotogales (tt. petrophila, tt. lettingae, thermosipho melanesiensis, and fervidobacterium nodosum) and a comprehensive comparative analysis including the original t. maritima genome. while ribosomal protein genes strongly place thermotogal ... | 2009 | 19307556 |
| gene cloning and characterization of a novel thermophilic esterase from fervidobacterium nodosum rt17-b1. | a bioinformatic screening of the genome of the thermophilic bacterium fervidobacterium nodosum rt17-b1 for esterhydrolyzing enzymes revealed a putative bacterial esterase (fne) encoded by fond_1301 with typical gdsl family motifs. to confirm its putative esterase function, the fne gene was cloned, functionally expressed in escherichia coli, and purified to homogeneity. recombinant fne exhibited the highest esterase activity of 14,000 u/mg with p-nitrophenyl acetate (pnpc(2)) as substrate. the ca ... | 2010 | 20383468 |
| molecular characterization of the glucose isomerase from the thermophilic bacterium fervidobacterium gondwanense. | the gene coding for xylose isomerase from the thermophilic bacterium fervidobacterium gondwanense was cloned and overexpressed in escherichia coli. the produced xylose isomerase (xyla), which closely resembles counterparts from thermotoga maritima and t. neapolitana, was purified and characterized. it is optimally active at 70 degrees c, ph 7.3, with a specific activity of 15.0 u/mg for the interconversion of glucose to fructose. when compared with t. maritima xyla at 85 degrees c, a higher cata ... | 2010 | 20718290 |
| fervidobacterium changbaicum lip1: identification, cloning, and characterization of the thermophilic lipase as a new member of bacterial lipase family v. | a novel lipase gene encoded 315 amino acid residues was obtained using lipase-prospecting primers and genome walking from hyperthermophilic bacterium fervidobacterium changbaicum cbs-1. sequence alignment and phylogenetic analysis revealed this novel lipase is a new member of bacterial lipase family v. the recombinant enzyme f. changbaicum lipase 1 (fclip1) showed maximum activity at 78 °c and ph 7.8. it displayed extreme thermostability at 70 °c and was also stable across a wide ph range from 6 ... | 2010 | 21046373 |
| fervidobacterium riparium sp. nov., a novel thermophilic anaerobic cellulolytic bacterium isolated from a kunashir hot spring. | a novel obligately anaerobic, extremely thermophilic, organotrophic bacterium, strain 1445tt, was isolated from a hot spring of kunashir island (kuril islands, russia). cells were motile rods with dimensions of 0.4-0.5 x 1.0-3.0 µm. the temperature range for growth at ph 7.8 was from 46 to 80°c, with the optimum at 65 °c. the ph range for growth at 65 °c was from 5.7 to 9.0, with the optimum at ph 7.8. the growth was not observed at or below 40 and at or above 85 °c as well as at or below ph 5.4 ... | 2010 | 21169457 |
| microbial diversity with dominance of 16s rrna gene sequences with high gc contents at 74 and 98 °c subsurface crude oil deposits in japan. | we prepared dna from the production waters of oil deposits and wellheads of the high- and hypertemperature japanese oil wells #ar39 (depth, 1230 m; temperature, 74 °c; pressure, 2.92 mpa) and #sr123 (depth, 1687 m; temperature, 98 °c; pressure, 11.3 mpa) to detect indigenous bacterial and archaeal microorganisms. we used pcr to amplify the 16s rrna genes of microbial communities and characterized them based on their sequences. a few species of microorganisms with high gc contents were detected i ... | 2011 | 21223337 |
| phylogeny and molecular signatures for the phylum thermotogae and its subgroups. | thermotogae species are currently identified mainly on the basis of their unique toga and distinct branching in the rrna and other phylogenetic trees. no biochemical or molecular markers are known that clearly distinguish the species from this phylum from all other bacteria. the taxonomic/evolutionary relationships within this phylum, which consists of a single family, are also unclear. we report detailed phylogenetic analyses on thermotogae species based on concatenated sequences for many ribos ... | 2011 | 21503713 |
| quantitative investigation of non-hydrolytic disruptive activity on crystalline cellulose and application to recombinant swollenin. | for the efficient degradation and bioconversion of cellulosic biomass, it is important to efficiently disrupt and convert crystalline regions of cellulose into easily hydrolyzable regions than to simply hydrolyze cellulose. expansin-like proteins such as swollenins have disruptive functions on lignocellulose, including crystalline cellulose, via non-hydrolytic mechanisms. in this work, we produced the swollenin from trichoderma asperellum in escherichia coli. the recombinant protein was then ref ... | 2011 | 21732246 |
| Influence of the N-terminal peptide on the cocrystallization of a thermophilic endo-ß-1,4-glucanase with polysaccharide substrates. | It is well known that protein cocrystallization is affected by several parameters such as the ratio of the protein to the ligand, the reservoir solution, the pH and the temperature. Previously, spatial blocking by the N-terminus was observed in the active site in the crystal structure of the native protein of a thermostable endoglucanase from the thermophilic bacterium Fervidobacterium nodosum Rt17-B1 (FnCel5A). It was speculated that the N-terminal a-helix might form interactions with the subst ... | 2011 | 22102031 |
| crystal structure of hyperthermophilic endo-β-1,4-glucanase: implications for catalytic mechanism and thermostability. | endo-β-1,4-glucanase from thermophilic fervidobacterium nodosum rt17-b1 (fncel5a), a new member of glycosyl hydrolase family 5, is highly thermostable and exhibits the highest activity on carboxymethylcellulose among the reported homologues. to understand the structural basis for the thermostability and catalytic mechanism, we report here the crystal structures of fncel5a and the complex with glucose at atomic resolution. fncel5a exhibited a (β/α)8-barrel structure typical of clan gh-a of glycos ... | 2011 | 22128157 |
| a novel thermostable and glucose-tolerant β-glucosidase from fervidobacterium islandicum. | an open reading frame (orf) encoding the enzyme β-glucosidase from the extremely thermophilic bacterium fervidobacterium islandicum has been identified, cloned and sequenced. the bgl1a gene was cloned in a pet-blue1 vector and transformed in escherichia coli, resulting in high-level expression of β-glucosidase fibgl1a that was purified to homogeneity in a two-step purification. fibgl1a is composed of 459 amino acid residues and showed high homology to glycoside hydrolase family 1 proteins. it ex ... | 2011 | 22146852 |