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31.
R. Huber Josef Stöhr Sabine Hohenhaus Reinhard Rachel Siegfried Burggraf Holger W. Jannasch Karl O. Stetter 《Archives of microbiology》1995,164(4):255-264
From a hydrothermal vent site off the Mexican west coast (20°50′N, 109°06′W) at a depth of 2,600 m, a novel, hyperthermophilic,
anaerobic archaeum was isolated. Cells were round to slightly irregular cocci, 1.2–2.5 μm in diameter and were motile by means
of a tuft of flagella. The new isolate grew between 60 and 93°C (optimum: 85°C), from pH 3.5 to 9 (optimum: pH 6.7), and from
0.8 to 8% NaCl (optimum: 2%). The isolate was an obligate organotroph, using chitin, yeast extract, meat extract, and peptone
for growth. Chitin was fermented to H2, CO2, NH3, acetate, and formate. H2S was formed in the presence of sulfur. The chitinoclastic enzyme system was oxygen-stable, cell-associated, and inducible
by chitin. The cell wall was composed of a surface layer of hex- americ protein complexes arranged on a p6 lattice. The core
lipids consisted of glycerol diphytanyl diethers and acyclic and cyclic glycerol diphytanyl tetraethers. The G+C content was
46.5 mol%. DNA/DNA hybridization and 16S rRNA sequencing indicated that the new isolate belongs to the genus Thermococcus, representing a new species, Thermococcus chitonophagus. The type strain is isolate GC74, DSM 10152.
Received: 8 May 1995 / Accepted: 26 June 1995 相似文献
32.
Rolland JL Gueguen Y Persillon C Masson JM Dietrich J 《FEMS microbiology letters》2004,236(2):267-273
A PCR protocol was used to identify and sequence a gene encoding a DNA ligase from Thermococcus fumicolans (Tfu). The recombinant enzyme, expressed in Escherichia coli BL21(DE3) pLysS, was purified to homogeneity and characterized. The optimum temperature and pH of Tfu DNA ligase were 65 degrees C and 7.0, respectively. The optimum concentration of MgCl2, which is indispensable for the enzyme activity, was 2 mM. We showed that Tfu DNA ligase displayed nick joining and blunt-end ligation activity using either ATP or NAD+, as a cofactor. In addition, our results would suggest that Tfu DNA ligase is likely to use the same catalytic residues with the two cofactors. The ability for DNA ligases, to use either ATP or NAD+, as a cofactor, appears to be specific of DNA ligases from Thermococcales, an order of hyperthermophilic microorganisms that belongs to the euryarchaeotal branch of the archaea domain. 相似文献
33.
34.
Kotaro Suematsu Toshifumi Ueda Takashi Nakashima Yoshimitsu Kakuta 《Bioscience, biotechnology, and biochemistry》2013,77(6):952-959
The ribonuclease P (RNase P) proteins TkoPop5 and TkoRpp30, homologs of human Pop5 and Rpp30, respectively, in the hyperthermophilic archaeon Thermococcus kodakarensis were prepared and characterized with respect to pre-tRNA cleavage activity using the reconstitution system of the well-studied Pyrococcus horikoshii RNase P. The reconstituted particle containing TkoPop5 in place of the P. horikoshii counterpart PhoPop5 retained pre-tRNA cleavage activity comparable to that of the reconstituted P. horikoshii RNase P, while that containing TkoRpp30 instead of its corresponding protein PhoRpp30 had slightly lower activity than the P. horikoshii RNase P. Moreover, we determined crystal structures of TkoRpp30 alone and in complex with TkoPop5. Like their P. horikoshii counterparts, whose structures were solved previously, TkoRpp30 and TkoPop5 fold into TIM barrel and RRM-like fold, respectively. This finding demonstrates that RNase P proteins in T. kodakarensis and P. horikoshii are interchangeable and that their three-dimensional structures are highly conserved. 相似文献
35.
Singleton MR Taylor SJ Parrat JS Littlechild JA 《Extremophiles : life under extreme conditions》2000,4(5):297-303
The gene encoding pyrrolidone carboxyl peptidase (Pcp) has been cloned from the hyperthermophilic archaeon Thermococcus litoralis. The recombinant enzyme has been expressed in Escherichia coli, purified, and char-acterized. The T. litoralis Pcp demonstrates strong sequence homology to previously characterized bacterial Pcps. Some investigations have been carried
out on enzyme substrate specificity and stability.
Received: July 4, 2000 / Accepted: July 21, 2000 相似文献
36.
37.
Fujiwara S Yamanaka A Hirooka K Kobayashi A Imanaka T Fukusaki E 《Biochemical and biophysical research communications》2004,325(3):1066-1074
Enzyme characteristics of trans-prenyl diphosphate synthase (Tk-IdsA) from Thermococcus kodakaraensis, which catalyzes the consecutive trans-condensation of isopentenyl diphosphate (C(5)) units with allylic diphosphate, were examined. Product analysis revealed that Tk-IdsA is a bifunctional enzyme, farnesyl diphosphate (FPP, C(15))/geranylgeranyl diphosphate (GGPP, C(20)) synthase, and mainly yields both C(15) and C(20). The FPP/GGPP product ratio increases with the rise of the reaction temperature. The kinetic parameters obtained at 70 and 90 degrees C demonstrated that the rise of the temperature elevates the k(0) value for the C(10) allylic substrate to more than those for the C(5) and C(15) allylic substrates. These data suggest that Tk-IdsA contributes to adjust the membrane composition to the cell growth temperature by modulating its substrate and product specificities. Mutation study indicated that the aromatic side chain of Tyr-81 acts as a steric hindrance to terminate the chain elongation and defines the final product length. 相似文献
38.
Heejin Bae Kee Pum Kim Jong Il Lee Jae-Geun Song Eui-Joon Kil Joong Su Kim Suk-Tae Kwon 《Extremophiles : life under extreme conditions》2009,13(4):657-667
The family B DNA polymerase gene from the archaeon Thermococcus marinus (Tma) contains a long open reading frame of 3,939 bp that encodes 1,312 amino acid residues. The gene is split by one intervening
sequence that forms a continuous open reading frame with the two polymerase exteins. In this study, the Tma DNA polymerase gene both with (precursor form) and without (mature form) its intein was expressed in Escherichia coli, purified by heat treatment and HiTrap™ Heparin HP column chromatography and characterized. Primary sequence analysis of the
mature Tma polymerase showed high sequence identity with DNA polymerases in the genus Thermococcus. The expressed precursor form was easily spliced during purification steps. The molecular mass of the purified Tma DNA polymerases is about 90 kDa, as estimated by SDS-PAGE. Both Tma DNA polymerases showed the same properties. PCR performed with this enzyme was found to be optimal in the presence of 50 mM
Tris–HCl (pH 8.4), 40 mM KCl, 12.5 mM (NH4)2SO4, 2 mM MgCl2, 0.05% Triton X-100 and 0.0075% BSA. Furthermore, long-range PCR and time-saving PCR were performed using various specific
ratios of Taq and Tma DNA polymerases (Tma plus DNA polymerase). 相似文献
39.
Dye-linked l-proline dehydrogenase catalyzes the oxidation of l-proline in the presence of artificial electron acceptors such as 2, 6-dichloroindophenol and ferricyanide. The enzyme from the hyperthermophilic archaeon Thermococcus profundus was purified and characterized for the first time in archaea by Sakuraba et al. in 2001. In this study, cloning and sequencing analyses of the gene encoding the enzyme and functional analysis of the subunits were performed. The gene formed an operon that consisted of four genes, pdhA, pdhB, pdhF, and pdhX, which are tandemly arranged in the order of pdhA-F-X-B. SDS-PAGE analysis of the purified recombinant enzyme showed four different bands corresponding to (54 kDa), (43 kDa), (19 kDa), and (8 kDa) subunits encoded by pdhA, pdhB, pdhF, and pdhX, respectively, and the molecular ratio of these subunits was determined to be equal. This indicates that the enzyme consists of a heterotetrameric structure. Functional analysis of each subunit revealed that the subunit catalyzed the dye-linked l-proline dehydrogenase reaction by itself and that, unexpectedly, the subunit exhibited dye-linked NADH dehydrogenase activity. This is the first example showing the existence of a bifunctional dye-linked l-proline/NADH dehydrogenase complex. On the basis of genome analysis, similar gene clusters were observed in the genomes of Pyrococcus horikoshii, Pyrococcus abyssi, Pyrococcus furiosus, and Archaeoglobus fulgidus. These results indicate that the dye-linked l-proline dehydrogenase is a novel type of heterotetrameric amino acid dehydrogenase that might be widely distributed in the hyperthermophilic archaeal strain.Communicated by K. Horikoshi 相似文献
40.
A systematic survey for thermophilic fermentative bacteria and archaea in high temperature petroleum reservoirs 总被引:8,自引:0,他引:8
Gino S. Grassia Keith M. McLean Philippé Glénat John Bauld Alan J. Sheehy 《FEMS microbiology ecology》1996,21(1):47-58
Abstract: Production waters from 36 high temperature petroleum reservoirs were examined for the presence of thermophilic, fermentative microorganisms. The direct supplementation of production waters with glucose and either yeast extract, peptone, tryptone or casamino acid resulted in the isolation of thermophilic, fermentative microorganisms from 47% of the petroleum reservoirs examined. Three distinctive morphological groups were isolated from the production waters of petroleum reservoirs with depths ranging from 396–3048 metres, temperatures ranging from 21–130°C, salinities ranging from 2.8–128 gl−1 and pHs ranging from 6.0–8.5. Group 1 were pleomorphic rod-shaped bacteria, Group 2 were sheathed rod-shaped bacteria, and Group 3 were coccoid archaea. Partial characterisation of strains from one seawater-flooded petroleum reservoir and three non-waterflooded petroleum reservoirs tentatively identified some strains in Group 1 as members of the genera Thermoanaerobacter and Thermoanaerobacterium , Group 2 as members of the Thermotogales order, and Group 3 as members of the genus Thermococcus . Production water salinity determined the type of microorganisms that were isolated. Group 1 organisms were found primarily in petroleum reservoirs with salinities less than 30 g/l, while Group 2 and 3 organisms were found to dominate in more saline reservoirs. The successful isolation of thermophilic, fermentative microorganisms from petroleum reservoirs decreased significantly with increasing salinity and temperature. These findings support the existence of a deep biosphere where fermentative microorganisms are widespread. 相似文献