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41.
Acetate oxidation in a thermophilic anaerobic sewage-sludge digestor: the importance of non-aceticlastic methanogenesis from acetate 总被引:3,自引:0,他引:3
Abstract Acetate conversion to methane in a steady-state, thermophilic (60°C) anaerobic sewage-sludge digestor and in a thermophilic (60°C) acetate chemostat inoculated with anaerobic thermophilic sewage sludge, was investigated by use of radiotracer methodology. When the acetate pool in the sewage-sludge digestor was 1–2 mM, 4.1% of 2-labeled acetate was converted to CO2 . However, when acetate was consumed to less than 1.0 mM, prior to isotopic examinations, this increased to 14.1%. Microscopic observations showed a shift in the acetate-degrading populations during start-up of the acetate-limited chemostat inoculated from the sewage-sludge digestor. Large numbers of Methanosarcina -aggregates were seen during the first 100–150 days of operation, while Methanosaeta -like rods were not observed. The Methanosarcina -aggregates disappeared concurrently with a decrease in the acetate concentration to approx. 0.4 mM, and the culture consisted mainly of a large number of autofluorescent, short rods together with fewer and longer, non-fluorescent, rods. Non-aceticlastic oxidation of acetate to methane was the mechanism of the acetate conversion in the chemostat after 7 months of operation. Our results indicate that the concentration of acetate can influence the mechanism of acetate conversion during thermophilic anaerobic digestion of organic matter. 相似文献
42.
Archaea are prokaryotes but some of their chaperoning systems resemble those of eukaryotes. Also, not all archaea possess the stress protein Hsp70(DnaK), in contrast with bacteria and eukaryotes, which possess it without any known exception. Further, the primary structure of the archaeal DnaK resembles more the bacterial than the eukaryotic homologues. The work reported here addresses two questions: Is the archaeal Hsp70 protein a chaperone, like its homologues in the other two phylogenetic domains? And, if so, is the chaperoning mechanism of bacterial or eukaryotic type? The data have shown that the DnaK protein of the archaeon Methanosarcina mazei functions efficiently as a chaperone in luciferase renaturation in vitro, and that it requires DnaJ, and the other bacterial-type chaperone, GrpE, to perform its function. The M. mazei DnaK chaperone activity was enhanced by interaction with the bacterial co-chaperone DnaJ, but not by the eukaryotic homologue HDJ-2. Both the bacterial GrpE and DnaJ stimulated the ATPase activity of the M. mazei DnaK. The M. mazei DnaK-dependent chaperoning pathway in vitro is similar to that of the bacterium Escherichia coli used for comparison. However, in vivo analyses indicate that there are also significant differences. The M. mazei dnaJ and grpE genes rescued E.coli mutants lacking these genes, but E.coli dnaK mutants were not complemented by the M. mazei dnaK gene. Thus, while the data from in vitro tests demonstrate functional similarities between the M. mazei and E.coli DnaK proteins, in vivo results indicate that, intracellularly, the chaperones from the two species differ. 相似文献
43.
Group II chaperonins belong to the Hsp60 family occurring in archaea and eukaryotes. The archaeal chaperonins build the thermosome, which is similar to the eukaryotic CCT (chaperonin-containing TCP-1). Eukaryotes have eight subunits, and up until now, it was thought that archaea had between one and three subunits, depending on the species. We now report two novel subunits, termed Hsp60-4 and Hsp60-5, in the archaeon Methanosarcina acetivorans, which also has Hsp60-1, Hsp60-2, and Hsp60-3 with orthologs in Methanosarcinae. Hsp60-4 and Hsp60-5 occur only in M. acetivorans, which makes this organism unique in that it has the highest number of chaperonin subunits ever described for an archaeon. Evolutionary analysis suggests that either Hsp60-4 or Hsp60-5 paralogs have arisen by gene duplication with vastly increased accepted substitution rates or that they represent ancestral types found only in this species.Reviewing Editor: Dr. W. Ford Doolittle 相似文献
44.
Acetate-grown cells of Methanosarcina barkeri MS were found to form methane from H2:CO2 at the same rate as hydrogen-grown cells. Cells grown on acetate had similar levels of soluble F420-reactive hydrogenase I, and higher levels of cytochrome-linked hydrogenase II compared to hydrogen-grown cells. The hydrogenase I and II activities in the crude extract of acetate-grown cells were separated by differential binding properties to an immobilized Cu2+ column. Hydrogenase II did not react with ferredoxin or F420, whereas hydrogenase I coupled to both ferredoxin and F420. A reconstituted soluble protein system composed of purified CO dehydrogenase, F420-reactive hydrogenase I fraction, and ferredoxin produced H2 from CO oxidation at a rate of 2.5 nmol/min · mg protein. Membrane-bound hydrogenase II coupled H2 consumption to the reduction of CoM-S-S-HTP and the synthesis of ATP. The differential function of hydrogenase I and II is ascribed to ferredoxin-linked hydrogen production from CO and cytochrome b-linked H2 consumption coupled to methanogenesis and ATP synthesis, respectively. 相似文献
45.
46.
Suresh Bhosale Deepa Kshirsagar Prashant Pawar Tulsiram Yeole Dilip Ranade 《FEMS microbiology letters》1995,127(1-2):151-155
Abstract 5-Aminolevulinic acid dehydratase from the archaebacterium Methanosarcina barken resembles the mammalian and yeast enzymes in its activation by Zn2+ , whereas its activation by K+ resembles the characteristic of bacterial enzymes. This enzyme is activated with Ni2+ which is a component of F430 , a cofactor present mainly in methanogens. The M r of 280000 for the native enzyme and 30 000 ± 2000 for the individual subunit suggest that the enzyme is composed of eight apparently indentical subunits similar to mammalian and yeast enzymes. The enzyme has two pH optima, at 8.5 and 9.4. Higher levels of 5-aminolevulinic acid dehydratase in acetate-grown cells suggest the possibility that regulation and control of this enzyme could be different on various growth substrates. 相似文献
47.
Methanogenic archaea conserve energy for growth by reducing some one- and two-carbon compounds to methane and concomitantly generating an ion motive force. Growth of Methanosarcina acetivorans on carbon monoxide (CO) is peculiar as it involves formation of, besides methane, formate, acetate and methylated thiols. It has been argued that methane formation is partially inhibited under carboxidotrophic conditions and that the other products result from either detoxification of CO or from bypassing methanogenesis with other pathways for energy conservation. To gain a deeper understanding of the CO-dependent physiology of M. acetivorans we analyzed metabolite formation in resting cells. The initial rates of methane, acetate, formate, and dimethylsulfide formation increased differentially with increasing CO concentrations but were maximal already at the same moderate CO partial pressure. Strikingly, further increase of the amount of CO was not inhibitory. The maximal rate of methane formation from CO was approximately fivefold lower than that from methanol, consistent with the previously observed significant downregulation of the energy converting sodium-dependent methyltransferase. The rate of dimethylsulfide formation from CO was only 1–2% of that of methane formation under any conditions tested. Implications of the data presented for previously proposed pathways of CO utilization are discussed. 相似文献
48.
Methanosarcina lacustris sp. nov., a new psychrotolerant methanogenic archaeon from anoxic lake sediments 总被引:1,自引:0,他引:1
Simankova MV Parshina SN Tourova TP Kolganova TV Zehnder AJ Nozhevnikova AN 《Systematic and applied microbiology》2001,24(3):362-367
A new psychrotolerant methanogenic archaeon strain ZS was isolated from anoxic lake sediments (Switzerland). The cells of the organism were non-motile cocci, 1.5-3.5 microm in diameter. The cells aggregated and formed pseudoparenchyma. The cell wall was Gram-positive. The organism utilized methanol, mono-, di-, trimethylamine and H2/CO2 with methane production. The temperature range for growth was 1-35 degrees C with an optimum at 25 degrees C. The DNA G+C content of the organism was 43.4. mol%. Analysis of the 16S rRNA gene sequence showed that strain ZS was phylogenetically closely related to members of the genus Methanosarcina, but clearly differed from all described species of this genus (95.6-97.6% of sequence similarity). The level of DNA-DNA hybridization of strain ZS with Methanosarcina barkeri and Methanosarcina mazei was 15 and 31%, respectively. Based on the results of physiological and phylogenetic studies strain ZS can be assigned to a new species of the genus Methanasarcina. The name Methanosarcina lacustris sp. nov. is proposed. The type strain is ZS (= DSM 13486T, VKM B-2268). 相似文献
49.
50.
Schäfer I Rössle M Biuković G Müller V Grüber G 《Journal of bioenergetics and biomembranes》2006,38(2):83-92
The first low-resolution shape of subunit F of the A1AO ATP synthase from the archaeon Methanosarcina mazei Gö1 in solution was determined by small angle X-ray scattering. Independent to the concentration used, the protein is monomeric and has an elongated shape, divided in a main globular part with a length of about 4.5 nm, and a hook-like domain of about 3.0 nm in length. The subunit-subunit interaction of subunit F inside the A1AO ATP synthase in the presence of 1-ethyl-3-(dimethylaminopropyl)-carbodiimide EDC was studied as a function of nucleotide binding, demonstrating movements of subunits F relative to the nucleotide-binding subunit B. Furthermore, in the intact A1AO complex, crosslinking of subunits D-E, A-H and A-B-D was obtained and the peptides, involved, were analyzed by MALDI-TOF mass spectrometry. Based on these data the surface of contact of B-F could be mapped in the high-resolution structure of subunit B of the A1AO ATP synthase. 相似文献