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The microbial community in the soil was analyzed during four weeks of a continuous enrichment of structural chernozem soil samples with a 0.1% solution of carboxymethyl cellulose (CMC) under aerobic and semianaerobic conditions. During the first 14 d, the total amount of the aerobic and anaerobic, cellulose-degrading microorganisms increased significantly. Various metabolic pathways were u‘ed te decompose the substrate: diverse metabolic systems were activated and different groups of microorganisms preferred in dependence on the presence of oxygen or the source of mineral nitrogen. In the later phases of cultivation, a decrease in the concentration of zymogenous microflora and in the level of substrate mineralization was observed ovon though CM-cellulase activity remained high. During the fourth week of cultivation, a conspicuous increase in the numbers of oligothropic bacteria occurring in the colcnies of the microorganisms degrading cellulose was found. The representatives of prosthecobacteria (Caulobacter, Hyphomicrobium, Prosthecomicrobium spp.) andSeliberia sp. were thus identified. This “microflora of dispersion” attends the zymogenous microbes degrading CMC and indicates later phases of the process of decomposition. 相似文献
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Isolation and characterization of a large,neurite-associated glycoconjugate from neuroblastoma cells
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A high molecular weight glycoconjugate has been isolated from neurite-producing neuronal tumor cells in culture and has been designated as I(0) based on its elution characteristics in gel filtration chromatography. This molecule cannot be found in a variety of nonneuronal cells. I(0) is found in the substratum-attached material or cell fraction of neurite-producing neuroblastoma cells, depending upon culture conditions. It is found in the substratum-bound fraction of B104 rat neuroblastoma cells during serum starvation and in the EGTA-detached cell fraction of B104 cells grown in chemically defined N2 medium. It occurs only in the cell fraction of the human neuroblastoma line Platt. Examination of behavioral variants of the B104 rat line further strengthens the association of I(0) with neurite production; the constitutive neurite-producing E(R)B9 variant contains I(0) while the non-neurite-producing E(R)A11 variant does not. I(0) is large, eluting in the void volume of sepharose-CL2B columns. Radioiodination of intact cells with lactoperoxidase shows I(0) to be a cell surface component. Metabolic radiolabeling studies show that it contains a high proportion of polysaccharide to protein, does not contain mannose, and is unsulfated. Alkaline borohydride reduction release two size classes of large polysaccharide chain. The alkaline reduction results, along with the mannose incorporation studies, show the presence of O-glycosidic linkages and few, if any, N-linkages. Resistance to nitrous acid deamination, insensitivity to glycosaminoglycan lyases, and the absence of sulfation, indicate that I(0) does not contain the glycosaminoglycans hyaluronic acid, chondroitin-, dermatan-, or heparin- sulfates. Affinity column chromatography reveals high binding affinity of I(0) to polyornithine and no binding to gelatin (collagen) or the glycosaminoglycans hyaluronate and heparin. These studies describe a unique high molecular weight glycoconjugate on the surface of neurite-producing neuroblastoma cell lines from two species. 相似文献
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Deep-level diagnostic value of the rDNA-ITS region 总被引:14,自引:0,他引:14
The similarity of certain reported angiosperm rDNA internal transcribed
spacer (ITS) region sequences to those of green algae prompted our analysis
of the deep-level phylogenetic signal in the highly conserved but short
5.8S and hypervariable ITS2 sequences. We found that 5.8S sequences yield
phylogenetic trees similar to but less well supported than those generated
by a ca. 10-fold longer alignment from rDNA-18S sequences, as well as
independent evidence. We attribute this result to our finding that,
compared to 18S, the 5.8S has a higher proportion of sites subject to vary
and greater among-site substitution rate homogeneity. We also determined
that our phylogenetic results are not likely affected by intramolecular
compensatory mutation to maintain RNA secondary structure nor by evident
systematic biases in base composition. Despite historical homology, there
appears to be no ITS2 primary sequence similarity shared sufficient
similarity to cluster correctly on the basis of alignability. Our results
indicate that groups, however, share sufficient similarity to cluster
correctly on the basis of alignability. Our results indicate that ITS
region sequences can diagnose organismal origins and phylogenetic
relationships at many phylogenetic levels and provide a useful paradigm for
molecular evolutionary study.
相似文献
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Production of Serine Proteases by the Oyster Pathogen Perkinsus marinus (Apicomplexa) In Vitro 总被引:1,自引:1,他引:0
JEROME F. LA PEYRE DORIS Y. SCHAFHAUSER ESAM H. RIZKALLA MOHAMED FAISAL 《The Journal of eukaryotic microbiology》1995,42(5):544-551
ABSTRACT. Analysis of the cell-free supernatants of Perkinsus marinus cultures by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining revealed the presence of as many as 17 bands ranging in molecular weight from 239 to 32 kDa. These bands were not present in un-inoculated medium. Moreover, P. marinus produces extracellular proteins that possess proteolytic activities; the cell-free supernatants of P. marinus cultures could digest a variety of proteins including gelatin, casein, fibronectin and laminin. Oyster plasma was also digested by cell-free culture supernatants. The proteolytic activity in cell-free culture supernatants was detected 24 h post-inoculation, while no proteolytic activity could be detected in cell lysates. The proteolytic activities were characterized using substrate-impregnated sodium dodecylsulfate-polyacrylamide gels and had approximate molecular weights ranging from 55 to 35 kDa. The proteolytic activity of cell-free culture supernatants was inhibited by the serine protease inhibitors phenylmethylsulphonyl fluoride, 3,4-dichloroisocoumarin and soybean trypsin inhibitor. In contrast, inhibitors (i.e. trans-epoxysuccinyll-leucylamido(4-guanidino)-butane, 1, 10-phenanthroline, captopril, ethylenediaminetetracetic acid, pepstatin A or diazoacetyl-DL-norleucine methyl ester) from the other three classes of proteases had no effect. It was concluded that the P. marinus proteases in cell-free culture supernatants are serine proteases. 相似文献
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Saks V. A. Khuchua Z. A. Vasilyeva E. V. Belikova O. Yu Kuznetsov A. V. 《Molecular and cellular biochemistry》1994,133(1):155-192
The published experimental data and existing concepts of cellular regulation of respiration are analyzed. Conventional, simplified considerations of regulatory mechanism by cytoplasmic ADP according to Michaelis-Menten kinetics or by derived parameters such as phosphate potential etc. do not explain relationships between oxygen consumption, workload and metabolic state of the cell. On the other hand, there are abundant data in literature showing microheterogeneity of cytoplasmic space in muscle cells, in particular with respect to ATP (and ADP) due to the structural organization of cell interior, existence of multienzyme complexes and structured water phase. Also very recent experimental data show that the intracellular diffusion of ADP is retarded in cardiomyocytes because of very low permeability of the mitochondrial outer membrane for adenine nucleotidesin vivo. Most probably, permeability of the outer mitochondrial membrane porin channels is controlled in the cellsin vivo by some intracellular factors which may be connected to cytoskeleton and lost during mitochondrial isolation. All these numerous data show convincingly that cellular metabolism cannot be understood if cell interior is considered as homogenous solution, and it is necessary to use the theories of organized metabolic systems and substrate-product channelling in multienzyme systems to understand metabolic regulation of respiration. One of these systems is the creatine kinase system, which channels high energy phosphates from mitochondria to sites of energy utilization. It is proposed that in muscle cells feed-back signal between contraction and mitochondrial respiration may be conducted by metabolic wave (propagation of oscillations of local concentration of ADP and creatine) through cytoplasmic equilibrium creatine and adenylate kinases and is amplified by coupled creatine kinase reaction in mitochondria. Mitochondrial creatine kinase has experimentally been shown to be a powerful amplifier of regulatory action of weak ADP fluxes due to its coupling to adenine nucleotide translocase. This phenomenon is also carefully analyzed.It is easier to explain biochemistry in terms of transport than it is to explain transport in terms of biochemistry. P. Mitchell The Ninth Sir Hans Krebs Lecture, Dresden, July 2, 1978. 相似文献
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