六碳糖和五碳糖同步糖化与共发酵的数学模型

Reliable production of biofuels and specifically bioethanol has attracted a significant amount of research recently. Within this context,this study deals with dynamic simulation of bioethanol production processes and in particular aims at developing a mathematical model for describing simultaneous saccharification and co-fermentation (SSCF) of C6 and C5 sugars. The model is constructed by combining existing mathematical models for enzymatic hydrolysis and co-fermentation. An inhibition of ethanol on cellulose conversion is introduced in order to increase the reliability. The mathematical model for the SSCF is verified by comparing the model predictions with experimental data obtained from the ethanol production based on kraft paper mill sludge. When fitting the model to the data, only the yield coefficients for glucose and xylose metabolism were fine-tuned, which were found to be 0.43 g.g-1 (ethanol/glucose) and 0.35 g.g-1 (ethanol/xylose) respectively. These promising validation results encourage further model application to evaluate different process configurations for lignocellulosic bioethanol technology.     

Simultaneous bioconversion of cellulose and hemicellulose to ethanol

Lignocellulosic materials containing cellulose, hemicellulose, and lignin as their main constituents are the most abundant renewable organic resource present on Earth. The conversion of both cellulose and hemicellulose for production of fuel ethanol is being studied intensively with a view to develop a technically and economically viable bioprocess. The fermentation of glucose, the main constituent of cellulose hydrolyzate, to ethanol can be carried out efficiently. On the other hand, although bioconversion of xylose, the main pentose sugar obtained on hydrolysis of hemicellulose, to ethanol presents a biochemical challenge, especially if it is present along with glucose, it needs to be fermented to make the biomass-to-ethanol process economical. A lot of attention therefore has been focussed on the utilization of both glucose and xylose to ethanol. Accordingly, while describing the advancements that have taken place to get xylose converted efficiently to ethanol by xylose-fermenting organisms, the review deals mainly with the strategies that have been put forward for bioconversion of both the sugars to achieve high ethanol concentration, yield, and productivity. The approaches, which include the use of (1) xylose-fermenting yeasts alone, (2) xylose isomerase enzyme as well as yeast, (3) immobilized enzymes and cells, and (4) sequential fermentation and co-culture process are described with respect to their underlying concepts and major limitations. Genetic improvements in the cultures have been made either to enlarge the range of substrate utilization or to channel metabolic intermediates specifically toward ethanol. These contributions represent real significant advancements in the field and have also been adequately dealt with from the point of view of their impact on utilization of both cellulose and hemicellulose sugars to ethanol.     

Characterization of recombinant E. coli ATCC 11303 (pLOI 297) in the conversion of cellulose and xylose to ethanol

This work describes the characterization of recombinant Escherichia coli ATCC 11303 (pLOI 297) in the production of ethanol from cellulose and xylose. We have examined the fermentation of glucose and xylose, both individually and in mixtures, and the selectivity of ethanol production under various conditions of operation. Xylose metabolism was strongly inhibited by the presence of glucose. Ethanol was a strong inhibitor of both glucose and xylose fermentations; the maximum ethanol levels achieved at 37 degrees C and 42 degrees C were about 50 g/l and 25 g/l respectively. Simultaneous saccharification and fermentation of cellulose with recombinant E. coli and exogenous cellulose showed a high ethanol yield (84% of theoretical) in the hydrolysis regime of pH 5.0 and 37 degrees C. The selectivity of organic acid formation relative to that of ethanol increased at extreme levels of initial glucose concentration; production of succinic and acetic acids increased at low levels of glucose (< 1 g/l), and lactic acid production increased when initial glucose was higher than 100 g/l.     

Changes in T-cell phenotype and adhesion molecules expression in psoriatic lesions after low-dose cyclosporin therapy

The fermentation of cellobiose is a rare trait among yeasts. Of the 308 yeast species that utilize cellobiose aerobically, only 12 species ferment it, and only 2 species, Candida molischiana and Candida wickerhamii, also ferment cellodextrins. Candida molischiana produced beta-glucosidase activity on all carbon sources tested, except glucose, mannose, and fructose. When these sugars were added to cultures growing on cellobiose, the synthesis of beta-glucosidase ceased. However, the total amount of enzyme activity remained constant, indicating that the C. molischiana beta-glucosidase is catabolite repressed and not catabolite inactivated. When grown in medium initially containing glucose plus xylose, cellobiose, maltose, mannitol, or glucitol, C. molischiana preferentially utilized glucose and produced little beta-glucosidase activity until glucose was nearly depleted from the medium. When grown in medium containing cellobiose plus either fructose or mannose, the yeast preferentially utilized the monosaccharides and produced little beta-glucosidase activity. Candida molischiana produced beta-glucosidase and co-utilized cellobiose and xylose, maltose, or trehalose. Glucose and fructose, mannose, or trehalose were co-utilized; however, no beta-glucosidase activity was detected. Thus, the order of substrate preference groups appeared to be (glucose, trehalose, fructose, mannose) > (cellobiose, maltose, xylose) > (mannitol, glucitol).     

Malignant melanoma in Europe: changes in mortality rates (1970-90) in European Community countries

We measured metabolic rates (mL O2 h-1, converted to kcal d-1), deep body temperatures (degree C), and skin temperatures (degree C) and calculated whole-animal thermal conductances (mL O2 g-1 h-1 degree C-1) of five 3-yr-old harbor seals (Phoca vitulina concolor) at air temperatures between -20 degrees and 35 degrees C. The mean thermal neutral zone of these seals extended from a lower critical temperature of -12.9 degrees +/- 1.6 degrees C (SD) to an upper critical temperature of 28.6 degrees +/- 1.7 degrees C. Hyperthermia was observed at an ambient air temperature of 35 degrees C. Mean standard metabolic rate was 1,553.6 +/- 168.2 kcal d-1, about 1.2 times the value expected for adult animals of similar body mass (mean mass = 49.2 +/- 7.5 kg). Mean deep body temperature increased from 37.5 degrees +/- 0.30 degrees C at an ambient temperature of 30 degrees C and reached 39.3 degrees +/- 0.33 degrees C at an ambient temperature of 35 degrees C. Skin temperature decreased with decreasing ambient temperature but remained well above ambient temperature. Mean whole-animal thermal conductance decreased from an ambient temperature of 35 degrees C until it reached a minimum value of 0.007 mL O2 g-1 h-1 degree C-1 at -4.0 degrees C; it then increased with a further decrease in ambient temperature. In comparison to the thermal limits of the same seals during their first year of life, the results indicate a broadening of the thermal neutral zone with age: an 11 degrees C decrease in the lower critical temperature and a 3.5 degrees C increase in the upper critical temperature. These findings suggest that warm ambient air temperatures should not pose any particular thermoregulatory problems for larger and older harbor seals, even beyond the limits of their current annual distribution.     

The vitamin B 6 requirements of the laying hen for reproduction

Some parameters related to withstanding severe cold (-20 degrees C) after administration of increasing doses of ethanol were investigated using guinea-pigs. The animals had been reared either at 22-23 degrees C or at 17-18 degrees C. They received ethanol in doses of 0.8 g, 1.2 g, 1.4 g or 1.6 g per kg of body weight. The fall of rectal temperature and its level at death were registered. The survival time, ethanol concentration in the blood and the brain, serum glucose and serum FFA at death were determined. In the animals reared at 22-23 degrees C the ethanol doses of 1.2-1.6 g/kg caused a significant shortening of the survival time and accelerated the fall of the rectal temperature. In addition the rectal temperature at death after ethanol was lower than in the controls. In the animals reared at 17-18 degrees C the ethanol doses used did not have any significant effect on the survival time and the rectal temperature. In both groups, ethanol concentration in the brain was lower (about 20-40%) than in the blood, the difference being greater in the group reared at 22-23 degrees C. Ethanol had no effect on the glucose concentrations. Serum FFA levels were slightly lower in animals reared at 22-23 degrees C than in those grown in the cooler temperature. It became evident that ethanol has a dose dependent deleterious effect on the thermoregulation of animals reared in warm (22-23 degrees C). The effect was seen at and above the dose of 1.2 g/kg. The results indicate further even a slight acclimation to cold was able to abolish the effect of these rather great doses of ethanol in severe cold exposure.     

Development of ascending colon in human fetal period

Study of glucose and xylose utilization by Pichia stipitis in a limited oxygen supply condition revealed that the rate of glucose utilization decreased rapidly while that of xylose decreased slowly until the time that glucose and xylose were shown to level out, at which point the rate of xylose utilization increased rapidly. Based on the results, ethanol fermentation technology in continuous connective tower fermenters was advanced, e.g., fermentation by P. stipitis in an airlift loop tower focusing on xylose utilization and then residue glucose utilization by Saccharomyces cerevisiae in an overflow tower. When the fed hydrolyzate of bagasse was concentrated in five folds and the dilution rate was 0.1 h-1, the total utilization ratio of reducing sugar was 97.2%; the concentration of ethanol was 46.4 g/L.h.     

Agar immobilized yeast cells in tubular reactor for ethanol production

Saccharomyces cerevisiae cells were immobilized in agar gel and used in a tubular reactor for conversion of cane molasses to ethanol at 30 degrees C, pH 4.5. Reactor was used in a continuous operation to test the operational stability and ethanol productivity. After 100 days of continuous fermentation at a dilution rate of 0.67 hr-1, some deactivation of cells was observed, but ethanol productivity was recovered by reactivating the cells by sparging air intermittently. It was found that intermittent reactivation during continuous operation was very important for satisfactory performance of the reactor. During operation, gel beads maintained their rigidity. Maximum ethanol concentration (94.9 g/L) was obtained with a feed containing 255 g/L reducing sugar, at a dilution rate of 0.2 hr-1. Maximum volumetric productivity (79.5 g ethanol /L/hr), specific ethanol productivity (0.58 g ethanol/g cells/hr), specific sugar uptake rate (1.12 g sugar/g cells/hr) and ethanol yield coefficient (0.43 g ethanol/g sugar) were obtained with a feed containing 195 g/L reducing sugar at a dilution rate of 1.33 hr-1.     

Glucocorticoids increase retinoid-X receptor alpha (RXRalpha) expression and enhance thyroid hormone action in primary cultured rat hepatocytes

A dynamic model of glucose overflow metabolism in batch and fed-batch cultivations of Escherichia coli W3110 under fully aerobic conditions is presented. Simulation based on the model describes cell growth, respiration, and acetate formation as well as acetate reconsumption during batch cultures, the transition of batch to fed-batch culture, and fed-batch cultures. E. coli excreted acetate only when specific glucose uptake exceeded a critical rate corresponding to a maximum respiration rate. In batch cultures where the glucose uptake was unlimited, the overflow acetate made up to 9. 0 +/- 1.0% carbon/carbon of the glucose consumed. The applicability of the model to dynamic situations was tested by challenging the model with glucose and acetate pulses added during the fed-batch part of the cultures. In the presence of a glucose feed, E. coli utilized acetate 3 times faster than in the absence of glucose. The cells showed no significant difference in maximum specific uptake rate of endogenous acetate produced by glucose overflow and exogenous acetate added to the culture, the value being 0.12-0.18 g g-1 h-1 during the entire fed-batch culture period. Acetate inhibited the specific growth rate according to a noncompetitive model, with the inhibition constant (ki) being 9 g of acetate/L. This was due to the reduced rate of glucose uptake rather than the reduced yield of biomass.     

Isolation and characterization of an extracellular glycosylated protein complex from Clostridium thermosaccharolyticum with pectin methylesterase and polygalacturonate hydrolase activity

An extracellular protein complex was isolated from the supernatant of a pectin-limited continuous culture of Clostridium thermosaccharolyticum Haren. The complex possessed both pectin methylesterase (EC 3.1.1.11) and exo-poly-alpha-galacturonate hydrolase (EC 3.2.1.82) activity and produced digalacturonate from the nonreducing end of the pectin chain. The protein consisted of 230- and 25-kDa subunits. The large subunit contained 10% (wt/wt) sugars (N-acetylgalactosamine and galactose). Under physiological conditions both activities acted in a coordinated manner: the ratio between methanol and digalacturonate released during degradation was constant and equal to the degree of esterification of the pectin used. Prolonged incubation of the enzyme with pectin led to a nondialyzable fraction that was enriched in neutral sugars, such as arabinose, rhamnose, and galactose; the high rhamnose/galacturonic acid ratio was indicative of hairy region-like structures. The smallest substrate utilized by the hydrolase was a tetragalacturonate. Vmax with oligogalacturonates increased with increasing chain length. The Km and Vmax for the polygalacturonate hydrolase with citrus pectate as a substrate were 0.8 g liter-1 and 180 mumol min-1 mg of protein-1, respectively. The Km and Vmax for the esterase with citrus pectin as a substrate were 1.2 g liter-1 and 440 mumol min-1 mg of protein-1, respectively. The temperature optima for the hydrolase and esterase were 70 and 60 degrees C, respectively. Both enzyme activities were stable for more than 1 h at 70 degrees C. The exo-polygalacturonate hydrolase of Clostridium thermosulfurogenes was partially purified while the methylesterase was also copurified.     

Factors related to Periotest values in endosseal implants: a 9-year follow-up

The lactose-negative yeast Rhodotorula glutinis 22P and the homofermentative lactic acid bacterium Lactobacillus helveticus 12A were cultured together in a cheese whey ultrafiltrate containing 42 g L-1 lactose. The chemical composition of the caroteno-protein has been determined. The carotenoid and protein contents are 248 micrograms g-1 dry cells and 48.2% dry weight. Carotenoids produced by Rhodotorula glutinis 22P have been identified as beta-carotene 15%, torulene 10%, and torularhodin 69%. After separating the cell mass from the microbial association, the exopolysaccharides synthesized by Rhodotorula glutinis 22P were isolated from the supernatant medium in a yield of 9.2 g L-1. The monosaccharide composition of the synthesized biopolymer was predominantly D-mannose (57.5%).     

Energetics and product formation by Saccharomyces cerevisiae grown in anaerobic chemostats under nitrogen limitation

Anaerobic fermentation of glucose (20 g/l) by Saccharomyces cerevisiae CBS 8066 was studied in a chemostat (dilution rate = 0.05-0.25 h-1) at different concentrations of the nitrogen source (5.00 g/l or 0.36 g/l ammonium sulphate). The ethanol yield (g ethanol produced/g glucose consumed) was found to be higher and the glycerol yield (g glycerol formed/g glucose consumed) lower during nitrogen limitation than under carbon limitation. The biomass yield on ATP (g dry weight biomass produced/mol ATP consumed), was consequently found to be lower during nitrogen-limited conditions.     

Microdialysis probe for transcutaneous monitoring of ethanol and glucose in humans

A heated (42 degrees C) microdialysis probe and its application for continuous transcutaneous sampling of ethanol and glucose through cellophane-stripped forearm skin are described. Ethanol and glucose concentration in the dialysate were measured on-line with continuous-flow analysis and compared with blood values in human volunteers after ethanol consumption (n = 4) and oral glucose testing (n = 5), respectively. For ethanol and glucose, the dialysate and blood concentrations were linearly related in each subject (r > or = 0.91, P < 0.005), although the dialysate-to-blood ratio varied among subjects. The recovery in vivo was 22.4 +/- 22.7 and 4.7 +/- 2.3% (SD) of the recovery in vitro for ethanol and glucose, respectively. The dialysate glucose concentration was independent of blood flow. When the probe temperature was increased from 32 to 42 degrees C, the dialysate-to-blood glucose ratio increased, with 2.4 +/- 1.4%/degrees C (SD) in fasting subjects (n = 4), which was similar to an increase of 2.1 +/- 0.045%/degree C in dialysate-to-medium ratio in vitro. The present approach for transcutaneous sampling may possibly be used for other substances of low molecular weight as well.     

Success rate of cytogenetic analysis at the time of second-trimester dilation and evacuation

Hypoxia elicits a number of compensatory responses in animals, including behavioral hypothermia. The hypothesis that hypoglycemia induces hypothermia in the bullfrog Rana catesbeiana was tested and that this behavioral response would be beneficial. Frogs equipped with a temperature probe were tested in a thermal gradient (10-40 degrees C). Insulin (15 IU kg-1) caused significant reduction of body temperature, from 25.0 to 17.8 degrees C. A non-metabolizable glucose analogue, 2-deoxy-D-glucose (2-DG, 50 mg kg-1), which blocks intracellular glucose utilization, was also injected and caused a similar drop in body temperature, despite an increase in plasma glucose levels. To assess the possible benefits of hypoglycemia-induced hypothermia, the effects of insulin and 2-DG injections were measured on plasma glucose concentration and on oxygen consumption of frogs equilibrated at 10, 20 and 30 degrees C. The plasma glucose was elevated at higher temperatures and so was oxygen consumption. The insulin caused a significant reduction of plasma glucose concentration (about 1.22 muMol ml-1) whereas 2-DG caused a significant increase (about 0.70 muMol ml-1) at 30 degrees C. Both drugs caused a reduction of oxygen consumption (approximately 0.388 and 0.382 ml min-1 kg at 30 degrees C after insulin and 2-DG injection, respectively). No effect of either insulin or 2-DG was observed when the animals were equilibrated at 10 degrees C. In conclusion, hypothermia may be a beneficial response to hypoglycemia in frogs.     

Power Doppler findings in nasopharingeal angiofibroma

Three fractions of polysaccharides that have high biological activity were extracted from Chinese bamboo leaves (Indocalamus Tesselatus) with 0.9% sodium chloride, 1% ammonium oxalate and 85% ethanol. Their component sugars were investigated by paper chromatography developed with n-butanol-pyridine-water (6:4:3 by vol), and by gas chromatography with crown ester stationary-phase-fused silica capillary column. Their monosaccharide compositions are different from each other with glucuronic acid, galactose, arabinose, mannose, glucose, and xylose (52, 18, 9, 7 and 6%, respectively) in one fraction, only glucuronic acid in another and glucuronic acid, fucose, arabinose and xylose (36, 31, 24 and 9%, respectively) in the third fraction. The protection effect of two polysaccharides against experimental hepatitis was studied in mice.     

Sugar composition of dietary fibre and short-chain fatty acid production during in vitro fermentation by human bacteria

The aim of the present study was to assess the relationship between the disappearance of dietary fibre sugars and the production of individual short-chain fatty acids (SCFA). The bacterial degradation of five dietary fibres whose sugars were quantified was investigated in vitro using a human faecal inoculum. Involvement of the main fibre sugars in SCFA production was evaluated by a stepwise multiple linear regression. The results show first that the nature and chiefly the associations between the fibre sugars were key variables in the fermentability. Second, the nature and the amounts of SCFA produced were closely related to the in vitro fermentation of the main sugars available: uronic acids seemed to be principally involved in the production of acetic acid whereas the production of propionic acid could be promoted by the fermentation of glucose and, to a lesser extent, by that of xylose and arabinose. Xylose tended to have a greater impact than uronic acids and glucose on the production of butyric acid. Thus, it would be possible to predict which SCFA could be specifically produced during the fermentation of a fibre, as far as the chemical composition and structure of this fibre are known.     

Inhibition of ethanol production by Saccharomyces cerevisiae in human blood by sodium fluoride

Production of ethanol in antemortem blood samples inoculated with an efficient ethanol-producing microorganism and incubated at various temperatures is discussed. Whole blood samples inoculated with Saccharomyces cerevisiae were incubated in gray stoppered Venoject tubes (approximate draw volume 7 mL) containing sodium fluoride (17.5 mg) and potassium oxalate (14.0 mg) at 4 degrees C, 25 degrees C, and 37 degrees C for 0, 24, 96, 192, and 408 h. No volatile substances (such as ethanol, methanol, isopropanol, acetone, or acetaldehyde) (< 0.010 g/dL) were produced in any of the samples at 4 or 25 degrees C. At 24 h incubation a trace amount (< 0.018 g/dL) of ethanol was detected at 37 degrees C.     

Use of almond mesocarp for production of the entomopathogenic fungus Verticillium lecanii

Almond mesocarp (AM) has been evaluated for production of Verticillium lecanii. Microbial flora of AM has been studied. After ground AM dilution plating, 5.3 x 10(5) +/- 2.6 x 10(4) fungal CFU.g-1 of dry AM were found. Common fungal saprophytes (Rhizopus sp. and Alternaria spp.) were found in more than 80% of the samples. Aspergillus sp. and yeasts were found less commonly. Rhizopus sp., Alternaria spp., and Aspergillus sp. inhibited growth of several V. lecanii; therefore, AM was treated with sterilization agents to eliminate endogenous mycoflora. Small samples (10 g) of AM saturated in distilled water treated with steam (120 degrees C and 100 kPa) were completely sterilized after 15 min. Ground AM incorporated on agar increased the biomass of V. lecanii compared with controls. This suggested AM as suitable substrate for the production of the fungus. In petri dishes, 9.7 x 10(7) +/- 2.9 x 10(7) conidia.g-1 of dry AM were produced after inoculating 10 conidia.g-1 of AM and incubating for 2 weeks. Viability of conidia produced was more than 90%. These conidia (5 x 10(4) conidia per larvae) caused Galleria mellonella mortality, calculated as median lethal time (LT50 5.3 +/- 1.6 days), that was significantly higher (F = 10.93; P < 0.05) than untreated controls (LT50 11.3 +/- 1.1 days). Larger scale tests have to be optimized before mass production.     

Training effects on maternal and fetal glucose uptake following acute exercise in the rat

The purpose of the present study was to determine the effects of chronic maternal exercise on glucose uptake in maternal tissues after one bout of treadmill running during late gestation in the rat and to determine the effects on glucose accumulation in the fetus and placenta. Trained pregnant animals (PR) ran at 30 m.min-1, on a 10 degrees incline for 1 h on day 20 of gestation with a similarly treated trained nonpregnant group (NPR). Immediately after the run the animals were infused with a bolus of 1 g.kg-1 body wt as a 50% dextrose solution mixed with 2-deoxy-D-[1-3H] glucose through a carotid catheter. Sedentary pregnant (P) and nonpregnant animals (C) were also infused with the solution after no food and water for the same time frame. After 60 min, tissues were analyzed for radioactivity. Radioactive tracer was augmented in the red gastrocnemius and soleus of the PR group and the soleus of P rats. However, tracer accumulation in the fetus and placenta of the trained animals was not different than P animals. These results indicate that acute exercise in trained animals increased glucose uptake in maternal skeletal muscles without compromising conceptus glucose accumulation.