D2O effects on association behavior and binding properties of phospho- and dephospho-cytoplasmic malic dehydrogenase

Changes in the concentration of three major carbohydrates, e.g., glycogen, trehalose, and cellulose, were determined during differentiation of Dictyostelium discoideum in a stage study. These three carbohydrates consituted 50–63% of the total carbohydrates. Total carbohydrate content per cell aliquot did not change between the aggregation and sorocarp stages of differentiation. The isolation, purification, and characterization of cellulose is described. Cellulose consisted of an alkali-insoluble (alpha) and an alkali-soluble (beta) fraction. Total cellulose accumulated from very low amounts in late pseudoplasmodium cells to about 35% of carbohydrates in mature sorocarps at a rate of 0.07 μmole glucose equiv/min/ml packed cell volume. Purified alkali-insoluble cellulose constituted about 19% of total carbohydrates in mature sorocarps and accumulated at a rate of 0.035 μmole glucose equiv/min/ml packed cell volume. Trehalose constituted 10–11% of the carbohydrates in sorocarps and accumulated at a rate of 0.035 μmole glucose equiv/min/ml packed cell volume. Glycogen, comparing several methods of determination, was rapidly degraded between the culmination and sorocarp stages of differentiation at an average rate of 0.05 μmole glucose equiv/min/ml packed cell volume. The major portion of glycogen was soluble in TCA and constituted 35% of total carbohydrates in aggregated cells and about 11% in mature sorocarps. A minor fraction of glycogen, about 15% of total carbohydrates in aggregated cells, was solubilized by KOH from a TCA precipitate. A mild acidic treatment of solubilized cell constituents increased the glycogen content by 55%, as judged by an enzymatic assay.     

Release of vasoactive intestinal peptide in the dumping syndrome.

To determine the effect of gastric surgery on the plasma vasoactive intestinal peptide (VIP) concentration, 13 patients with gastrectomy and seven controls were given an oral hypertonic load (200 ml 50% glucose solution). Blood was taken at intervals during the test for measurement of VIP and blood glucose concentrations and packed cell volume. At the same time observations were made on the occurrence of dumping symptoms and a record kept of the pulse rate. VIP values in the patients with gastrectomy were significantly increased by glucose ingestion, while these did not alter in controls (p less than 0.001). There was a highly significant correlation between the rate of rise in plasma VIP concentration and the rates of rise in packed cell volume (r = 0.85; p less than 0.001) and blood glucose concentration (r = 0.76; p less than 0.01) in patients with gastrectomy. Changes in packed cell volume and blood glucose values and the occurrence of dumping symptoms during the test were significantly different when postoperative patients were compared with controls (p less than 0.001, p less than 0.005, and p less than 0.001 respectively). Furthermore, when the patients with gastrectomy were divided into those without symptoms and those with dumping after meals the latter group showed a significantly greater rise of VIP (p less than 0.05). Despite the increased plasma VIP concentrations observed during dumping, VIP cannot be taken as the sole factor in the pathogenesis of the dumping syndrome.     

Some blood parameters of the rainbow trout (Salmo gairdneri Richardson)

Fifty rainbow trout of the Kamloops strain were examined for 12 haematological parameters: erythrocyte sedimentation rate, haemoglobin, packed cell volume, erythrocyte count, erythrocyte diameters, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, leukocyte count, differential leukocyte count, plasma total protein and plasma glucose concentration. The fish had been held under known environmental and dietetic conditions, and at the time of sampling were 14 months old. The majority of results for erythrocyte sedimentation rate, haemoglobin, packed cell volume, erythrocyte count, erythrocyte diameters, total protein and differential leukocyte count fell within narrow ranges. The total leukocyte counts and glucose levels were more widely spread. The results are discussed and compared with those already published for Idaho and Shasta strains. It is impossible to say whether the differences that were observed between Kamloops and these other varieties were due to strain alone, since other variables were present. Some problems associated with establishing normal ranges for these parameters in fish are discussed.     

Effect of various sources of organic carbon and high nitrite and nitrate concentrations on the selection of denitrifying bacteria. II. Continuous cultures in packed bed reactors

The effect of different organic compounds, nitrites and nitrates at the concentration of 1,000 mg N/l on the quantitative and strain-specific selection of denitrifying bacteria was determined in anaerobic packed bed reactors. Both the source of carbon and nitrogen form influenced strain specificity and the frequency of occurrence of denitrifying bacteria. The frequency of denitrifying bacteria within packed bed reactor ranged in different media from 11% (glucose and nitrates) to 100% (methanol and ethanol with nitrates). A single species selection was observed in the presence of nitrites within packed bed reactor: Pseudomonas aeruginosa in medium with acetate. Pseudomonas stutzeri in medium with ethanol, Pseudomonas mendocina in medium with methanol and Pseudomonas fluorescens in medium with glucose. When nitrates were present in packed bed reactor, the dominating bacteria were: P. stutzeri in medium with acetate, P. fluorescens in medium with ethanol, Paracoccus denitrificans in medium with methanol and Alcaligenes faecalis in medium with glucose.     

In vitro studies of whole blood and buffycoat-free packed cells in CPD-adenine with 1.5-fold glucose concentration and 0.4 mm adenine (author's transl)]

We studied a new CPD-adenine preservative with 1.5-fold of usual glucose concentration and 0.4 mM adenine during 6 weeks of storage in whole blood and buffycoat-free packed cells. Measurements of ATP and glucose indicated no necessity for increasing the glucose concentration in the standard CPD-adenine.     

Immobilized glucoamylase on porous glass

A study was made to determine the controlling mass transfer resistance in the overall reaction rate for conversion of maltose to glucose, catalyzed by glucoamylase immobilized onto porous glass. For normal operation of a packed column and air-stirred batch reactor, the rate controlling step was found to be the internal resistance of simultaneous pore diffusion and chemical reaction. Experimental effectiveness factors were determined and are compared with those derived from a theoretical diffusion model based on Michaelis-Menten kinetics. Also given are temperature and pH relationships for the free and immobilized glucoamylase.     

Cellobiose hydrolysis using Pichia etchellsii cells immobilized in calcium alginate

The rate of celluose degradation, limited due to the inhibition by cellobiose, can be increased by the hydrolysis of cellobiose to glucose using immobilized beta-glucosidase. Production of beta-glucosidase in four yeasts was studied and a maximum activity of 1.22 IU/mg cells was obtained in cells of Pichia etchellsii when grown on 3% cellobiose as the sole carbon source. A study of the immobilization of beta-glucosidase containing cells of Pichia etchellsii on various solid supports was conducted and immobilization by entrapment in calcium alginate gel beads was found to be the most simple and efficient method. A retention of 96.5% of initial activity after ten sequential batch uses of the immobilized preparation was observed. The pH and temperature optima for free and immobilized cells were the same, i.e., 6.5 (0.05M Maleate buffer) and 50 degrees C, respectively. Even though the temperature optimum was found to be 50 degrees C, the enzyme exhibits a better thermal stability at 45 degrees C. Beads stored at 4 degrees C for six months retain 80% of their activity. Kinetic studies performed on free and immobilized cells shown that glucose is a noncompetitive product inhibitor.The immobilized preparation was found to be limited by pore diffusion but exhibited no film-diffusion resistance during packed bed column indicated by a low dispersion number of 0.1348. A model for reaction with pore diffusion for a noncompetitive type of inhibited system was developed and applied to the cellobiose hydrolysis system. The rate of reaction with diffusional limitations was determined by using the model and effectiveness factors were calculated for different particle sizes. An effectiveness factor of 0.49 was obtained for a particle diameter of 2.5 mm. The modified rate expression using the effectiveness factor represented batch and packed bed reactor operation satisfactorily. The productivity in the packed bed column was found to fall rapidly with increase in conversion rate indicating that the operating conditions of the column would have to be a compromise between high conversion rates and reasonable productivity. A half-life of over seven days was obtained at the operating temperature of 45 degrees C in continuous operation of the packed bed reactor. However, the half-life in the column was found to be greatly affected by temperature, increasing to over seventeen days at a temperature of 40 degrees C and decreasing to less than two days at 50 degrees C.     

Viability, ATP and 2,3-DPG content of resuspended erythrocytes during several-week storage in cold

Leukocyte-and thrombocyte-poor packed red cells obtained from ACD or. ACD-AG blood were resuspended to a hematocrit of about 55% and stored at 4 degrees C. The resuspension solution consisted of xylitol, inorganic phosphate, bicarbonate, adenine (A) and guanosine (G) solved in water. In one case glucose, citrate and sucrose were also added, in another one, sorbitol. The 2,3-DPG and the ATP level remained for a longer period in the sorbitol-xylitol-medium than in the glucose-xylitol-medium. The ATP content in the red cell suspension was higher than in packed cells. Higher ATP values were obtained in red blood cells from whole blood with adenine and guanosine. The survival rate of resuspended red blood cells in glucose-AG-citrate-sucrose medium was about 80--85% after 3 weeks of storage and 77% after 6 weeks with a higher range.     

Mathematical modeling of ethanol production by immobilized Zymomonas mobilis in a packed bed fermenter

A mathematical model which describes ethanol production in a packed bed fermenter containing. Zymomonas mobilis entrapped in small spheres of calcium alginate within a packed bed fermenter has been developed. The equations combine simultaneous diffusion and reaction as well as a complex flow pattern to calculate glucose and ethanol profiles in the column type reactor. As part of the study, diffusivity values for glucose and ethanol in cell-loaded calcium alginate were determined. Also a freecell kinetic expression for Z. mobilis at 33 degrees C and ph 6.0 was developed. Comparison of the model with actual experimental results were made showing average deviations of ca. 30-40%.     

Ethanol production in a hollow fiber bioreactor using Saccharomyces cerevisiae

Summary A new approach for continuous production of ethanol was developed using a Hollow fiber fermentor (HFF). Saccharomyces cerevisiae cells were packed into the shell-side of a hollow fiber module. Using 100 g/l glucose in the feed gave an optimum ethanol productivity, based on total HFF volume, of 40 g ethanol/l/h at a dilution rate of 3.0 h^-1. Under these conditions, glucose utilization was 30%. However, at 85% glucose utilization the productivity was 10 g ethanol/l/h. This compares to batch fermentor productivity of 2.1 g ethanol/l/h at 100% glucose utilization.     

Biodegradation of Malathion with Indigenous Acclimated Activated Sludge in Batch Mode and in Continuous-Flow Packed-Bed Reactor

Acclimated activated sludge was examined for its ability to degrade malathion with and without the presence of glucose as a potential cometabolite substrate. In this study, a packed-bed reactor (PBR) using three kinds of biofilm carriers was employed for efficient degradation of malathion. The results obtained indicate that microorganisms tested were able to degrade malathion. The observed degradation rate of the pesticide in the presence of glucose was the same as without glucose. The activated sludge was found to be able to use malathion as the sole phosphorus source. In contrast, the degradation ability of the activated sludge was lost when the pesticide was used as the sole source of sulfur. The degradation capacity of the PBR was higher than the performance obtained with the batch reactor. The reactor packed with crushed olive kernels exhibited the best performance, allowing a total removal of malathion (10 mg/dm³) within 12 h.     

Optimum temperature operation mode for glucose isomerase reactor operating at constant glucose conversion

The optimum temperature operation mode required to achieve constant outlet glucose conversion is determined for immobilized glucose isomerase continuous packed bed reactor. The reactor design equation assumes reversible Michaelis-Menten kinetics with both enzyme deactivation and substrate protection. An increasing temperature profiles are determined for different operating periods, residence times and glucose conversions. The temperature increase with time is very small at low degree of glucose conversion and at relatively long residence time. The temperature rise with time increases at high degree of conversion and at relatively short residence time.     

Characteristics of a biofilm photobioreactor as applied to photo-hydrogen production

The application of a cell immobilization technique to a biofilm-based photobioreactor was developed to enhance its photo-hydrogen production rate and light conversion efficiency. Rhodopseudomonas palustris CQK 01 was initially attached to the surface of packed glass beads to form a biofilm in this experiment. Then, the biofilm photobioreactor (BPBR) was illuminated by light-emitting diodes with light wavelengths of 470, 590 and 630 nm and hydrogen was evolved with glucose being the sole carbon source. Under the illumination condition of 5000 lux illumination intensity and 590 nm wavelength, the BPBR showed good hydrogen production performance: the hydrogen production rate was 38.9 ml/l/h and light conversion efficiency was 56%, while the hydrogen yield was 0.2 mol H₂/mol glucose. Furthermore, results show that the highest hydrogen production rate and glucose removal rate were obtained when the glucose concentration is 0.12 M, the optimal pH 7 and optimal temperature of influent liquid 25 °C.     

EFFECT OF ATRAZINE ON CHLORELLA VULGARIS

Samples of a synchronous culture of Chlorella vulgaris were treated with: (a) atrazine, (b) atrazine + glucose, or (c) glucose. Cultures were sampled 0, 24, 48, and 72 hr, after treatment. Chlorophyll and packed cell volume were determined on each, and electron micrographs were taken. Atrazine prevented normal increase in packed cell volume and chlorophyll; this cessation of growth was counteracted by glucose. Chlorella utilized both the endogenous products of photosynthesis and an exogenous energy source, glucose in this instance, simultaneously. The atrazine-treated cells did not contain starch; control cells, atrazine-glucose- and glucose-treated cells did. Atrazine did not cause any observable abnormalities in cell organelles.     

Thermophilic biohydrogen production from glucose with trickling biofilter

Thermophilic H2 production from glucose was studied at 55-64 degrees C for 234 days using a continuous trickling biofilter reactor (TBR) packed with a fibrous support matrix. Important parameters investigated included pH, temperature, hydraulic retention time (HRT), and glucose concentration in the feed. The optimal pH and temperature were 5.5 and 60 degrees C, respectively. With decreasing HRT or increasing inlet glucose concentration, volumetric H2 production rate increased but the H2 production yield to glucose decreased gradually. The biogas composition was almost constant at 53 +/- 4% (v/v) of H2 and 47 +/- 4% (v/v) of CO2. No appreciable CH4 was detected when the reactor was under a normal operation. The carbon mass balance showed that, in addition to cell mass, lactate, n-butyrate, CO2, and acetate were major products that comprised more than 85% of the carbon consumed. The maximal volumetric H2 production rate and H2 yield to glucose were 1,050 +/- 63 mmol H2/l.d and 1.11 +/- 0.12 mol H2/mol glucose, respectively. These results indicate that the thermophilic TBR is superior to most suspended or immobilized reactor systems reported thus far. This is the first report on continuous H2 production by a thermophilic TBR system.     

A comparative study of immobilized amyloglucosidase in a packed bed reactor and a continuous feed stirred tank reactor

The catalytic activity of amyloglucosidase covalently attached to DEAE-cellulose was studied in a packed bed reactor and a continuous feed stirred tank reactor (CSTR) for the reaction maltose → glucose. At low flow rates mass-transfer limitations in the bed reactor lead to lower conversions for this reactor compared to the CSTR. Simple theoretical expressions for these reactors were compared with the experimental results. There are significant differences between the kinetic parameters and pH profile of the immobilized and free enzyme. The immobilized enzyme also showed greater stability at 50°C than did free amyloglucosidase. The temperature dependence of the reaction rate was the same for immobilized and free enzyme.     

Parameter oscillation attenuation and mechanism exploration for continuous VHG ethanol fermentation

A bioreactor system composed of a stirred tank and three tubular bioreactors in series was established, and continuous ethanol fermentation was carried out using a general Saccharomyces cerevisiae strain and a very high gravity medium containing 280 g L(-1) glucose, supplemented with 5 g L(-1) yeast extract and 3 g L(-1) peptone. Sustainable oscillations of glucose, ethanol, and biomass were observed when the tank was operated at the dilution rate of 0.027 h(-1), which significantly affected ethanol fermentation performance of the system. After the tubular bioreactors were packed with 1/2' Intalox ceramic saddles, the oscillations were attenuated and quasi-steady states were achieved. Residence time distributions were studied for the packed bioreactors by the step input response technique using xylose as a tracer, which was added into the medium at a concentration of 20 g L(-1), indicating that the backmixing alleviation assumed for the packed tubular bioreactors could not be established, and its contribution to the oscillation attenuation could not be verified. Furthermore, the role of the packing's yeast cell immobilization in the oscillation attenuation was investigated by packing the tubular bioreactors with packings with significant difference in yeast cell immobilization effects, and the experimental results revealed that only the Intalox ceramic saddles and wood chips with moderate yeast cell immobilization effects could attenuate the oscillations, and correspondingly, improved the ethanol fermentation performance of the system, while the porous polyurethane particles with good yeast cell immobilization effect could not. And the viability analysis for the immobilized yeast cells illustrated that the extremely lower yeast cell viability within the tubular bioreactors packed with the porous polyurethane particles could be the reason for their inefficiency, while the yeast cells loosely immobilized onto the surfaces of the Intalox ceramic saddles and wood chips could be renewed during the fermentation, guaranteeing their viability and making them more efficient in attenuating the oscillations. The packing Raschig rings without yeast cell immobilization effect did not affect the oscillatory behavior of the tubular bioreactors, further supporting the role of the yeast cell immobilization in the oscillation attenuation.     

Monitoring of temperature effects on animal cell metabolism in a packed bed process

Animal cell (Chinese Hamster Ovary) concentration was determined on-line in a packed bed process using dielectric spectroscopy. This enabled the evaluation of the effect of temperature on specific metabolic rates during 3 months of continuous culture. The effect of low cultivation temperature on cell growth and metabolism was monitored, and the data were used for process development. At 37 degrees C cells grew exponentially with a specific growth rate of 0.038 d-1 and specific glucose uptake and lactate production rates increased continually. Reduction of the temperature to 33.5 degrees C resulted in a lowering of these metabolic rates while having no effect on cell proliferation. Subsequent reduction of the temperature to 32 degrees C resulted in stabilization of the cell concentration at a high density (3.6 x 10(7) cell per mL of packed bed). In addition, the specific production rate of the protein of interest increased by a factor of 6 compared to the value at 37 degrees C. During the stationary phase at 32 degrees C, all other specific metabolic rates could be controlled to low and constant levels.     

Ethanol production in an immobilized-cell reactor

Biomass can be converted to sugars by hydrolysis with enzymes or mineral acids. These sugars can be converted into a number of chemical intermediates in biological reactors. Biological reactions are generally slow and selection of the most efficient reactor is important in these applications. Immobilized-cell reactors allow high cell densities and high throughput by attaching the microorganisms to a fixed support. This paper examines the rate of production of ethanol from glucose by Saccharomyces cerevisia in a packed column. These rates are compared with those for the same reaction in a stirred reactor.     

Continuous ethanol production by thermotolerant Kluyveromyces marxianus IMB3 immobilized on mineral Kissiris at 45°C

Thermotolerant Kluyveromyces marxianus var. marxianus IMB3 yeast strain was immobilized on Kissiris (mineral glass foam derived from lava) in column packed reactors, and used for ethanol production from glucose or molasses under continuous culture conditions at temperatures between 40 and 50°C. Both ethanol yield and fermentation efficiency were highest at 45°C and a dilution rate (D) of 0.15/h. Increasing sugar concentration led to an increase in ethanol yield of up to 68.6 and 55.9 g/l on approx. 200g glucose or molasses, respectively. Optimum fermentation efficiency (experimental yields over theoretical maximum yields) however was at about 15% sugar for both glucose and molasses. Slight aeration (25 ml of air/min) through the medium addition line was found advantageous due to its mixing effect and probable maintenance of activity.