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11.
Metabolism at the cytosol–mitochondria interface and its regulation is of major importance particularly for efficient production of biopharmaceuticals in Chinese hamster ovary (CHO) cells but also in many diseases. We used a novel systems-oriented approach combining dynamic metabolic flux analysis and determination of compartmental enzyme activities to obtain systems level information with functional, spatial and temporal resolution. Integrating these multiple levels of information, we were able to investigate the interaction of glycolysis and TCA cycle and its metabolic control. We characterized metabolic phases in CHO batch cultivation and assessed metabolic efficiency extending the concept of metabolic ratios. Comparing in situ enzyme activities including their compartmental localization with in vivo metabolic fluxes, we were able to identify limiting steps in glycolysis and TCA cycle. Our data point to a significant contribution of substrate channeling to glycolytic regulation. We show how glycolytic channeling heavily affects the availability of pyruvate for the mitochondria. Finally, we show that the activities of transaminases and anaplerotic enzymes are tailored to permit a balanced supply of pyruvate and oxaloacetate to the TCA cycle in the respective metabolic states. We demonstrate that knowledge about metabolic control can be gained by correlating in vivo metabolic flux dynamics with time and space resolved in situ enzyme activities. 相似文献
12.
《Journal of biological dynamics》2013,7(2):674-694
This paper analyses a bionomic model of two competitive species in the presence of toxicity with different harvesting efforts. An interesting dynamics in the first quadrant is analysed and two saddle-node bifurcations are detected for different bifurcation parameters. It is noted that under certain parametric restrictions, the model has a unique positive equilibrium point that is globally asymptotically stable whenever it is locally stable. It is also noted that the model can have zero, one or two feasible equilibria appearing through saddle-node bifurcations. The non-existence of a limit cycle in the interior of the first quadrant is also discussed using the Poincare–Dulac criteria. The saddle-node bifurcations are studied using Sotomayor's theorem. Numerical simulations are carried out to validate the analytical findings. The conditions for the existence of bionomic equilibria are discussed and an optimal harvesting policy is derived using Pontryagin's maximum principle. 相似文献
13.
T.S. Nedeva V.A. Savov A.V. Kujumdzieva-Savova E.R. Davidov 《FEMS microbiology letters》1993,107(1):49-52
Abstract A screening procedure for highly thermostable yeast superoxide dismutase was developed. Growth yields at various temperatures were estimated for ten mesophilic and thermotolerant strains, belonging to the genera Saccharomyces, Kluyveromyces and Pichia . Higher yields at 45°C were obtained for K. lactis 90-3 and 90-4. A correlation between the ability to grow at higher temperature and the thermostability of the superoxide dismutase enzyme synthesized was observed. A comparison of the operational stability of the superoxide dismutase of all tested strains suggests that the enzyme of K. lactis strains was more thermostable than that of the other tested microorganisms. 相似文献
14.
Nitric oxide (NO) is a chemical weapon within the arsenal of immune cells, but is also generated endogenously by different bacteria. Pseudomonas aeruginosa are pathogens that contain an NO-generating nitrite (NO2−) reductase (NirS), and NO has been shown to influence their virulence. Interestingly, P. aeruginosa also contain NO dioxygenase (Fhp) and nitrate (NO3−) reductases, which together with NirS provide the potential for NO to be metabolically cycled (NO→NO3−→NO2−→NO). Deeper understanding of NO metabolism in P. aeruginosa will increase knowledge of its pathogenesis, and computational models have proven to be useful tools for the quantitative dissection of NO biochemical networks. Here we developed such a model for P. aeruginosa and confirmed its predictive accuracy with measurements of NO, O2, NO2−, and NO3− in mutant cultures devoid of Fhp or NorCB (NO reductase) activity. Using the model, we assessed whether NO was metabolically cycled in aerobic P. aeruginosa cultures. Calculated fluxes indicated a bottleneck at NO3−, which was relieved upon O2 depletion. As cell growth depleted dissolved O2 levels, NO3− was converted to NO2− at near-stoichiometric levels, whereas NO2− consumption did not coincide with NO or NO3− accumulation. Assimilatory NO2− reductase (NirBD) or NorCB activity could have prevented NO cycling, and experiments with ΔnirB, ΔnirS, and ΔnorC showed that NorCB was responsible for loss of flux from the cycle. Collectively, this work provides a computational tool to analyze NO metabolism in P. aeruginosa, and establishes that P. aeruginosa use NorCB to prevent metabolic cycling of NO. 相似文献
15.
Michel Fons Brigitte Cami Jean-Claude Patte Marc Chippaux 《Molecular & general genetics : MGG》1987,206(1):141-143
Summary A library of Deusulfovibrio desulfuricans Norway genomic DNA was constructed in Escherichia coli with pBR322 as vector and plasmids able to complement the proA and leuB mutations of the host were screened. It was observed that all the plasmids studied were highly unstable, the insert DNA being rapidely lost under non-selective growth conditions. A 2.75 kb DNA fragment of D. desulfuricans Norway was found to complement E. coli ProA, ProB and ProC deficiencies. From the results of restriction analysis and Southern hybridization, it is proposed that the genes involved in proline and leucine biosynthesis are clustered on the chromosome of D. desulfuricans Norway. 相似文献
16.
17.
Effect of roasting on ochratoxin A level in green coffee beans inoculated with Aspergillus ochraceus
Haruo Tsubouchi Katsuhiko Yamamoto Kazuo Hisada Yoshio Sakabe Shun- ichi Udagawa 《Mycopathologia》1987,97(2):111-115
The heat stability of ochratoxin A in green coffee beans inoculated with Aspergillus ochraceus was studied. Heat treatment (roasting) at 200 °C for 10 or 20 min reduced the levels of ochratoxin A by only 0–12% in the dried whole beans. Almost all of the ochratoxin A was infused into the coffee decoction when the roasted samples were ground and extracted with boiling water. Therefore, the reduction of ochratoxin A concentration of contaminated coffee beans by roasting under these conditions is ineffective. 相似文献
18.
Francisco Javier Caballero Isabel Igeño Jacobo Cárdenas Francisco Castillo 《Archives of microbiology》1989,152(5):508-511
The phototrophic bacterium Rhodobacter capsulatus E1F1 assimilates ammonia and other forms of reduced nitrogen either through the GS/GOGAT pathway or by the concerted action of l-alanine dehydrogenase and aminotransferases. These routes are light-independent and very responsive to the carbon and nitrogen sources used for cell growth. GS was most active in cells grown on nitrate or l-glutamate as nitrogen sources, whereas it was heavily adenylylated and siginificantly repressed by ammonium, glycine, l-alanine, l-aspartate, l-asparagine and l-glutamine, under which conditions specific aminotransferases were induced. GOGAT activity was kept at constitutive levels in cells grown on l-amino acids as nitrogen sources except on l-glutamine where it was significantly induced during the early phase of growth. In vitro, GOGAT activity was strongly inhibited by l-tyrosine and NADPH. In cells using l-asparagine or l-aspartate as nitrogen source, a concerted induction of l-aspartate aminotransferase and l-asparaginase was observed. Enzyme level enhancements in response to nitrogen source variation involved de novo protein synthesis and strongly correlated with the cell growth phase.Abbreviations ADH
l-alanine dehydrogenase
- AOAT
l-alanine:2-oxoglutarate aminotransferase
- Asnase
l-asparaginase
- GOAT
Glycine: oxaloacetate aminotransferase
- GOGAT
Glutamate synthase
- GOT
l-aspartate: 2-oxoglutarate aminotransferase
- GS
Glutamine synthetase
- HPLC
High-Pressure Liquid Chromatography
- MOPS
2-(N-morpholino)propanesulfonic acid
- MSX
l-methionine-d,l-sulfoximine 相似文献
19.
A natural coordinate system is introduced for the analysis of the global stability of the Hardy-Weinberg (HW) polymorphism under the general multilocus additive viability model. A global convergence criterion is developed and used to prove that the HW polymorphism is globally stable when each of the loci is diallelic, provided the loci are overdominant and the multilocus recombination is positive. As a corollary the multilocus Hardy-Weinberg law for neutral selection is derived.Research supported in part by NIH grants GM 39907-01, GM 10452-26 and NSF Grant DMS 86-06244Research supported in part by a US-Israel Binational Science Foundation grant 85-00021 and NIH grant GM 28016 相似文献
20.
An S I R S epidemiological model with vital dynamics in a population of varying size is discussed. A complete global analysis is given which uses a new result to establish the nonexistence of periodic solutions. Results are discussed in terms of three explicit threshold parameters which respectively govern the increase of the total population, the existence and stability of an endemic proportion equilibrium and the growth of the infective population. These lead to two distinct concepts of disease eradication which involve the total number of infectives and their proportion in the population.Partially supported by NSF Grant No. DMS-8703631. This work was done while this author was visiting the University of VictoriaResearch supported in part by NSERC A-8965 相似文献