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A newly isolated strain of Cunninghamella echinulata grown on glucose produced significant quantities of biomass and cellular lipids in media with high C/N ratio. The oil yield from glucose consumed increased after nitrogen exhaustion in the growth medium, but gamma-linolenic acid (GLA) content in cellular oil systematically decreased during the lipid accumulation process. When lipid accumulation was completed, GLA concentration in the cellular lipids progressively increased. The highest GLA production (720 mg/l) was achieved in medium with a C/N ratio equal to 163. C. echinulata was also able to grow on orange peel. The C/N ratio in the orange peel decreased from 50 to 26 during solid-state fermentation. Maximum oxygen uptake was observed during assimilation of reducing sugars, whereas a polygalacturonase activity was detected after reducing sugars had been exhausted. The maximum GLA production was 1.2-1.5 mg/g of fermented peel, calculated on a dry weight basis. After enrichment of the pulp with inorganic nitrogen and glucose, an increase in the production of oil and GLA was observed.
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PMID:Production of gamma-linolenic acid by Cunninghamella echinulata cultivated on glucose and orange peel. 1193 80

Because pigments of phototrophs can be involved either in photosynthesis or photoprotection, pigmentation changes in response to nutrient availability can affect how cells interact with their solar environment. We investigated the impact of nitrogen availability both on pigmentation of the cyanobacterium Synechococcus sp. strain SH-94-5 and on carbon assimilation by this strain in the presence or absence of UV radiation. Pigmentation changes in strain SH-94-5 due to ammonium exhaustion included phycobiliprotein degradation, an exponential decline in chlorophyll a content, and a net increase in beta-carotene. Following its replenishment, ammonium stimulated non-photosynthetic carbon assimilation for several hours prior to the resumption of photosynthesis and growth. Carbon fixation during this lag phase was concurrent with the metabolism of glycogen reserves, and it is likely that inorganic carbon was incorporated into glycogen-derived carbon skeletons primarily for amino acid synthesis. In contrast, carbon fixation was almost exclusively photosynthetic during exponential growth. UV-A radiation (320-400 nm) inhibited photosynthetic but not non-photosynthetic carbon assimilation. Only growing cells were inhibited, and the disappearance of inhibition following nitrogen depletion appeared to result from the reduction of cellular photosensitizing targets below a threshold level rather than from the inactivation of photosynthesis.
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PMID:Effects of nitrogen availability on pigmentation and carbon assimilation in the cyanobacterium Synechococcus sp. strain SH-94-5. 1197 48

IOGEN Corporation of Ottawa, Canada, has recently built a 40t/d biomass-to-ethanol demonstration plant adjacent to its enzyme production facility. It has partnered with the University of Toronto to test the C6/C5 cofermenta-tion performance characteristics of the National Renewable Energy Labora-tory's metabolically engineered Zymomonas mobilis using various biomass hydrolysates. IOGEN's feedstocks are primarily agricultural wastes such as corn stover and wheat straw. Integrated recombinant Z. mobilis strain AX101 grows on D-xylose and/or L-arabinose as the sole carbon/energy sources and ferments these pentose sugars to ethanol in high yield. Strain AX101 lacks the tetracycline resistance gene that was a common feature of other recombinant Zm constructs. Genomic integration provides reliable cofermentation performance in the absence of antibiotics, another characteristic making strain AX101 attractive for industrial cellulosic ethanol production. In this work, IOGEN's biomass hydrolysate was simulated by a pure sugar medium containing 6% (w/v) glucose, 3% xylose, and 0.35% arabinose. At a level of 3 g/L (dry solids), corn steep liquor with inorganic nitrogen (0.8 g/L of ammonium chloride or 1.2 g/L of diammonium phosphate) was a cost-effective nutritional supplement. In the absence of acetic acid, the maximum volumetric ethanol productivity of a continuous fermentation at pH 5.0 was 3.54 g/L x h. During prolonged continuous fermentation, the efficiency of sugar-to-ethanol conversion (based on total sugar load) was maintained at >85%. At a level of 0.25% (w/v) acetic acid, the productivity decreased to 1.17 g/L x h at pH 5.5. Unlike integrated, xylose-utilizing rec Zm strain C25, strain AX101 produces less lactic acid as byproduct, owing to the fact that the Escherichia coli arabinose genes are inserted into a region of the host chromosome tentatively assigned to the gene for D-lactic acid dehydrogenase. In pH-controlled batch fermentations with sugar mixtures, the order of sugar exhaustion from the medium was glucose followed by xylose and arabinose. Both the total sugar load and the sugar ratio were shown to be important determinants for efficient cofermentation. Ethanol at a level of 3% (w/v) was implicated as both inhibitory to pentose fermentation and as a potentiator of acetic acid inhibition of pentose fermentation at pH 5.5. The effect of ethanol may have been underestimated in other assessments of acetic acid sensitivity. This work underscores the importance of employing similar assay conditions in making comparative assessments of biocatalyst fermentation performance.
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PMID:Performance testing of Zymomonas mobilis metabolically engineered for cofermentation of glucose, xylose, and arabinose. 1201 70

The purpose of this investigation was to determine the influence of post-exercise macronutrient intake on weight loss, protein metabolism, and endurance exercise performance during a period of increased training volume. Ten healthy young female endurance athletes performed 4 60-min bouts of cycle ergometry at approximately 65% of VO2peak on 4 days (day 1, 3, 4, and 6) during 2 separate 1-week periods. On day 7, participants performed a ride to exhaustion at approximately 75% of VO2peak. One of the 7-day periods served as a control condition, where a placebo beverage was consumed following the exercise bouts on days 1, 3, 4, and 6 (CON). During the other 7-day protocol (POST), participants consumed a predefined formula beverage with added carbohydrate following the exercise bouts on days 1, 3, 4, and 6. Energy intake and macronutrient proportions were the same between the 2 trials; the only difference was the timing at which the macronutrients were consumed. Calculated fat oxidation was greater during exercise on day 6 during POST as compared to CON (p < .05). Glucose and insulin concentrations were significantly higher (p < .05) following exercise during POST as compared to CON. There was a trend (p = .06) for nitrogen balance to be greater on days 5 and 6 with POST as compared to CON. Time to exhaustion during exercise on day 7 was longer during POST as compared to CON (p < .05). POST resulted in a maintenance of body weight during the 7-day protocol, while there was a significant (p < .05) reduction with CON. It was concluded that post-exercise macronutrient intake following endurance exercise can attenuate reductions in body weight and improve nitrogen balance during 7 days of increased energy expenditure. Importantly, post-exercise supplementation improved time to exhaustion during a subsequent bout of endurance exercise.
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PMID:The influence of post-exercise macronutrient intake on energy balance and protein metabolism in active females participating in endurance training. 1218 17

A suitable strain and important factors influencing citric acid formation in yeasts were identified. Candida oleophila ATCC 20177 was chosen as the best citric acid producer from several Candida strains. Yields of 50 g/l citric acid were produced in shake flask and 80 g/l in fed-batch fermentations with 1.5 and 3 g/l NH(4)Cl under non-optimized conditions. Ammonium nitrogen was identified as the limiting substrate for citrate formation. Citric acid excretion begins a few hours after exhaustion of nitrogen in the medium. The importance of intracellular nitrogen limitation was clarified by elemental analysis of C. oleophila biomass. The nitrogen content of C. oleophila biomass decreased from 7.45% during the growth phase to 3.96% in the production phase. The biomass contained less carbon and more trace elements in the growth phase compared with the production phase. Relatively high intracellular NH(4)(+) concentration of about 1.2 mg/g biomass (~37.4 mM) was found during the production phase. The low intracellular nitrogen content and increase of intracellular NH(4)(+) concentration, possibly caused by proteolysis following extracellular nitrogen exhaustion, trigger citric acid production. Intracellular nitrogen limitation and the increase in intracellular NH(4)(+) concentration are the most important factors influencing citric acid formation in yeasts.
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PMID:Citric acid production by Candida strains under intracellular nitrogen limitation. 1238 45

The nitrogenase (acetylene reductase) activity in monolithic and minced peat samples was found to be low, no more than 0.014-0.022 mg N/(kg h). Incorporation of the 15N2 isotope into organic compounds of peat soil was from 2.71-8.13 mg N/kg over 15 days. The nitrogen-fixing activity was the highest in a 10-20 cm layer of soil and much lower in the upper (under green moss) and deeper (20-30 cm) layers. The addition of glucose to soil samples stimulated nitrogen fixation considerably after 18-26 h. The maximum nitrogenase activity (3.5-3.8 mg N/(kg h)) observed after 60-70 h coincided with the peak of respiratory activity. A repeated addition of glucose after its exhaustion increased nitrogenase activity without a lag period to 8.5 mg N/(kg h). Investigation of the effect of environmental factors (temperature, pH, aeration, and light intensity) on potential nitrogen-fixing activity in peat samples revealed that nitrogen fixation could proceed in a wide range of pH values (from 3.0 to 7.5) and temperatures (from 5 to 35 degrees C). The nitrogen-fixing bacteria belonging to different trophic groups were enumerated by using nitrogen-free media with pH values and mineralization levels close to those in situ. In samples of peat soil, diazotrophic methanol-utilizing bacteria prevailed (2.0-2.5 x 10(6) cells/g); the second largest group was facultatively anaerobic bacteria of the family Enterobacteriaceae.
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PMID:[Nitrogen-fixing activity in peat soils from a raised bog]. 1269 1

The carbon and nitrogen mineralisation of a composting mixture of brewing yeast and lemon tree prunings was studied, at different degrees of stabilisation of this matrix, within an incubation experiment in soil. Meanwhile, a growth test in pots with ryegrass (Lolium perenne L.) was carried out using the selected soil and equal amounts of the composting mixture taken at different maturation steps, in order to evaluate the additions of these organic amendments in terms of fertilising value. Samples of the composting mixture, when poorly transformed through the biostabilisation process, showed high CO2-C releases in the soil, due to the microbial attack on easily degradable organic fractions still present in the mixture, with 24.7% mineralisation of the initial total organic carbon (TOC) after a 70 day incubation. On the other hand, mature compost was the most stable matrix, with only 5.4% of TOC mineralised after 70 days. Furthermore, amendments with the initial composting mixture led to negative net N-mineralisation during 56 days of incubation with soil. Only slight negative values of the net N-mineralisation were detected with fully stabilised compost. Nevertheless, pot experiments with ryegrass revealed that mature compost may promote N mineralisation to certain extents. Moreover, mature compost did not produce any phytotoxic effect, behaving as a slow-action organic fertiliser with N made available through a progressive mineralisation. Thus, the results gained through this study are a confirmation that the fertilising quality of a compost destined for agricultural uses is heavily affected by the complete exhaustion of the maturation reactions.
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PMID:Carbon mineralisation and plant growth in soil amended with compost samples at different degrees of maturity. 1273 31

Baarda, I. F. (Iowa State University, Ames) and W. R. Lockhart. Characteristics and significance of the physiological patterns accompanying growth limitation. J. Bacteriol. 84:1085-1093. 1962.-The physiological characteristics of several related strains of Escherichia coli were studied during the transition from exponential growth to stationary phase in defined media with limiting concentrations of the carbon or nitrogen source. The metabolic changes typical of this transition, which often are set in motion before exhaustion of the limiting nutrient has occurred, appear to consist of a series of shifts in the relative emphasis placed on competing reaction sequences leading, respectively, to cell division, protoplasmic synthesis, or cell maintenance. This process of biochemical differentiation has characteristic patterns which are qualitatively distinct for particular limiting nutrients and quantitatively different among strains. Correlation of specific metabolic patterns with the characteristic values of certain growth constants for each strain permits hypotheses concerning the physiological significance of these constants.
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PMID:Characteristics and significance of the physiological patterns accompanying growth limitation. 1396 12

Freter, Rolf (Jefferson Medical College, Philadelphia, Pa.), and Atsushi Ozawa. Explanation for limitation of populations of Escherichia coli in broth cultures. J. Bacteriol. 86:904-910. 1963.-Veal Infusion Broth surrounding a cellophane bag containing a 24-hr culture of Escherichia coli was inhibitory to an inoculum of the same strain when kept under an atmosphere of nitrogen plus CO(2). The inhibition could be abolished by addition of glucose or by aeration. When a small inoculum of a dulcitol-positive E. coli strain was introduced into a fully grown (24 hr) static Veal Infusion Broth culture of a dulcitol-negative E. coli, no multiplication occurred. However, the inoculum did grow in the presence of a carbon source (dulcitol) which could not be utilized by the static population. The logarithmic growth rate attained by the dulcitol-positive inoculum under these conditions was independent of the dulcitol concentration. In contrast, the maximal population size was a function of the amount of dulcitol present. Similar results were obtained when sucrose or salicin was substituted for the dulcitol. All strains grew well in filtrates of 24-hr broth cultures with growth rates and maximal populations approximating those attained in fresh broth. It was thus concluded that populations of E. coli in broth cultures were limited by exhaustion of all those carbon and energy sources which could be utilized under the prevailing highly reduced conditions. No evidence of inhibitory metabolic end products was found except in broth supplemented with 1% glucose. Partial inhibition of multiplication was noted in the latter medium even when the pH was maintained between 6.9 and 7.2.
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PMID:EXPLANATION FOR LIMITATION OF POPULATIONS OF ESCHERICHIA COLI IN BROTH CULTURES. 1408 Aug

Heterogeneous populations acclimated to sorbitol or mannitol were subjected to shock loading with glucose during growth on the sugar alcohol. Under these severe shock-loading conditions, glucose caused immediate cessation of sorbitol or mannitol removal. Metabolism of these compounds was renewed subsequent to glucose exhaustion. Under similar shock-loading conditions, with the use of cells operationally defined as "old," the sugar alcohols were removed concurrently with glucose. In the absence of a nitrogen source with old cells, the added glucose was removed slowly in the presence of sorbitol and not at all in the mannitol-acclimated system. These results indicate the potential variability in purification efficiency attributable to substrate interaction.
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PMID:SEQUENTIAL SUBSTRATE REMOVAL IN RESPONSE TO QUALITATIVE SHOCK LOADING OF ACTIVATED SLUDGE SYSTEMS. 1417 Sep 68


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