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Metabolism analysis and on-line physiological state diagnosis of acetone-butanol fermentation.

Fermentation equations for acetone-butanol (AB) were applied in a metabolic analysis of the reaction network under various conditions; that is, at different pHs and a high NADH2 turnover rate using methyl viologen, in a Clostridium acetobutylicum culture. The results disclosed variations in the pattern of rate changes that reflected changes in the physiological state. A linear relationship was found to exist between NADH2 generation and butanol production rate. By coupling an automated measurement system with the fermentation model, on-line estimation of the culture state was accomplished. Based on the AB fermentation model, new parameters were defined for on-line diagnosis of the physiological state and determination of the best timing for amplifying NADH2 generation by the addition of methyl viologen to obtain a high level of butanol productivity. A potential means of achieving optimal control for a high level of solvent production, involving the correlation of certain rates, is proposed.

Acetone↗

Sequential solubilization of proteins precipitated with trichloroacetic acid in acetone from cultured Catharanthus roseus cells yields 52% more spots after two-dimensional electrophoresis.

Sample preparation is still the most critical step in two-dimensional gel electrophoresis (2-DE), and needs to be optimized for each type of sample. To analyze the proteome of the medicinal plant Catharanthus roseus, we developed and evaluated a sequential solubilization procedure for the solubilization of proteins after precipitation in trichloroacetic acid and acetone. The procedure includes solubilization with a conventional urea buffer followed by a stronger solubilizing buffer containing thiourea. The sequential solubilization of the precipitated proteins results in very different spot patterns following 2-DE. The number of protein spots which could be detected in both samples of the sequential solubilization was only about 10% of the total number of spots. Compared to solubilization in a single step, the total number of spots that could be detected in the sequential solubilization procedure was increased by 52%. The method described is simple and is applicable to different types of plant tissue.

Acetone↗

Application of free-flow electrophoresis to the purification of trichosanthin from a crude product of acetone fractional precipitation.

The application of free-flow electrophoresis (FFE) to the purification of trichosanthin (TCS) from a crude product of acetone fractional precipitation was investigated. An electrophoresis technique, combining field step electrophoresis (FSE) and zone electrophoresis (ZE) to a one-step procedure, was optimized until a satisfactory purification factor (1.35), high resolution, and purity (>99%) were achieved. Testing several separation buffer systems revealed that a throughput of 14.2 mg/h can be obtained when the very basic TCS (pI 10.1) was dissolved and electrophoresed in a phosphate buffer system of pH 4. The purity of electrophoresed trichosanthin was proved by a variety of analytical methods, such as sodium dodecyl sulfate (SDS)-gel electrophoresis, capillary isoelectric focusing (CIEF), and sequencing of N- and C-termini. The high purity and large throughput achieved at low cost by using FFE indicates that this method can be employed for TCS purification.

Acetone↗

Effects of carbon tetrachloride, menadione, and paraquat on the urinary excretion of malondialdehyde, formaldehyde, acetaldehyde, and acetone in rats.

Excretions of the lipid peroxidation products, formaldehyde (FA), acetaldehyde (ACT), malondialdehyde (MDA), and acetone (ACON), were simultaneously identified and quantitated in the urine of female Sprague-Dawley rats by gas chromatography-mass spectroscopy (GC-MS) and high pressure liquid chromatography (HPLC) following the acute administration of carbon tetrachloride, a model alkylating agent that does not induce glutathione depletion, and the redox cycling compounds paraquat and menadione. All three xenobiotics are well-known inducers of oxidative stress. Oxidative stress was induced by oral administration of single doses of 2.5 mL of carbon tetrachloride/kg, 60 mg menadione/kg, and 75 mg paraquat/kg. These doses are approximately 50% of the LD50's for the three xenobiotics. Urinary excretion of FA, ACT, MDA, and ACON increased relative to control animals following treatment with all xenobiotics. Over the 48 hours of the study, the greatest increases in the excretion of MDA, FA, ACT, and ACON occurred after paraquat administration, with increases of approximately 2.7-, 2.6-, 4.3-, and 11.0-fold, respectively. This technique may have wide-spread applicability as an effective biomarker for investigating altered lipid metabolism in disease states and exposure to environmental pollutants/xenobiotics.

Acetaldehyde↗

The acetone soluble fraction from bark extract of Stryphnodendron adstringens (Mart.) coville inhibits gastric acid secretion and experimental gastric ulceration in rats.

The acetone soluble fraction from a crude methanol extract of Stryphnodendron adstringens stem bark (AFSAB) was evaluated in acute (ethanol, indomethacin and hypothermic restraint-stress) and chronic (acetic acid) models of gastric ulceration and on basal and bethanechol-stimulated gastric acid secretion in rats. Rats pretreated orally with AFSAB at doses of 400 and 800 mg/kg showed significant decreases of gastric lesion scores in ethanol (62% and 98%) and hypothermic restraint-stress (89% and 88%) models but exerted no significant influence on indomethacin-induced acute or acetic acid-induced chronic ulceration. In pylorus-ligated rats, AFSAB significantly decreased the basal as well as bethanechol-stimulated gastric secretory volume and the total acidity with an elevated pH value. AFSAB failed to modify the gastric mucus and the gastric wall nonprotein-sulphydryl content. These results point to a possible antisecretory effect of AFSAB which account for the observed antiulcer activity in ethanol and hypothermic restraint-stress induced models of acute gastric ulceration.

Acetone↗

Drosophila alcohol dehydrogenase activity in vitro and in vivo: effects of acetone feeding.

When adult Drosophila are placed on medium containing 0.5% acetone, their level of alcohol dehydrogenase activity drops rapidly. At the same time, the proportion of activity in the various electrophoretic forms of the enzyme shifts; most of the activity becomes localized in what is ordinarily a minor form of the enzyme. Moreover, the loss of enzyme activity occurs in vivo as well, as shown by sensitivity to ethanol poisoning, insensitivity to pentenol treatment, and inability to utilize ethanol as an energy source. These observations are discussed in light of a model advanced for the origin of the multiple forms of alcohol dehydrogenase in Drosophila.

Acetone↗

Excretion of malondialdehyde, formaldehyde, acetaldehyde, acetone and methyl ethyl ketone in the urine of rats given an acute dose of malondialdehyde.

A high pressure liquid chromatographic system (HPLC) has recently been developed for the simultaneous detection of malondialdehyde (MDA), formaldehyde (FA), acetaldehyde (ACT) and acetone (ACON). We have examined the urinary excretion of these four lipid metabolites in the urine of rats following the acute oral administration of MDA (158 mg/kg body weight). During the first 12 h, increases in the urinary excretion of MDA and ACT of approximately 192- and 70-fold, respectively, were observed. The urinary excretion of both MDA and ACT decreased thereafter. An increase in FA excretion was observed only 12-24 h after MDA administration. A significant decrease in ACON relative to control values was observed 12-48 h after MDA treatment. Two new peaks were present in the HPLC chromatograms of urine samples 0-24 h after MDA administration. Both peaks were shown to be due to methyl ethyl ketone (MEK) which appears to be formed as a result of MDA metabolism. The results demonstrate that orally administered MDA is rapidly excreted in the urine, and alters the metabolism and excretion of other lipid metabolites.

Acetaldehyde↗

Effects of crude adlay hull acetone extract on corticosterone release from rat zona fasciculata-reticularis cells.

Adlay is a grass crop which has been used in traditional Chinese medicine and also as a nourishing food. It has been shown to posses anti-allergic, antimutagenic and hypolipemic effects. However, the effects and action mechanisms of crude adlay hull acetone extract (AHA) on adrenal zona fasciculata-reticularis (ZFR) cells are still unclear. This study explored the effects of AHA on corticosterone release. ZFR cells were incubated with AHA in the presence or absence of adrenocorticotropin (ACTH), 8-bromo-cyclic 3': 5'- adenosine monophosphate (8-Br-cAMP), forskolin (FSK), 25-hydroxy cholesterol (25-OH-cholesterol), pregnenolone, progesterone or deoxycorticosterone. The concentrations of corticosterone or pregnenolone in the media were measured by radioimmunoassay (RIA). The cells were used to measure the expression of steroidogenic acute regulatory (StAR) protein by Western blot. The present data demonstrated that: (1) AHA inhibited ACTH-, 8-Br-cAMP-, forskolin-, 25-OH-cholesterol-, pregnenolone-, progesterone- or deoxycorticosterone-stimulated corticosterone release; (2) AHA (800 microg/ml) caused more pregnenolone release in control group, but not in 25-OH-cholesterol, trilostane or 25-OH-cholesterol+trilostane group; (3) kinetic study showed an uncompetitive inhibition model of AHA to P450 side chain cleavage enzyme (P450scc); (4) kinetic study showed a noncompetitive inhibition model of AHA to 11beta-hydroxylase; and (5) AHA inhibited the expression of StAR protein. These results suggest that AHA acts directly upon rat ZFR cells to diminish corticosterone release. These results indicate the inhibitory mechanism of AHA mediates through an inhibition of the activities of the post-cAMP corticosterone synthesis enzymes, i.e. 3beta-HSD, 21-hydroxylase, 11beta-hydroxylase, and inhibition of StAR protein expression.

3-Hydroxysteroid Dehydrogenases↗

Kinetic behaviors between acetone and composite bead in biofilter.

In this study, the kinetic behaviors between acetone and composite bead were investigated. The microbial growth rate decreased with increasing average inlet concentration and increased with increasing operation temperature at average inlet concentration ranging from 50 to 300 ppm and operation temperature ranging from 30 to 40 degrees C. The microbial growth rate would be inhibited with increasing average inlet concentration, and the inhibitive effect was more pronounced at higher operation temperature. The microbial growth rate would be enhanced with increasing operation temperature, and the enhancing effect was more pronounced at higher average inlet concentration. The values of maximum reaction rate Vm and half-saturation constant Ks ranged from 0.04 to 0.05 g-C/h-kg packing material and from 37.19 to 42.77 ppm, respectively. The biochemical reaction model could be regarded as the zero-order kinetic with the diffusion rate limitation. The biochemical reaction rate decreased with increasing average inlet concentration and increased with increasing operation temperature. The biochemical reaction rate would be inhibited with increasing average inlet concentration, and the inhibitive effect was more pronounced at lower operation temperature. The biochemical reaction rate would be enhanced with increasing the operation temperature, and the enhancing effect was more pronounced at higher average inlet concentration. The maximum elimination capacity of biofilter increased with increasing operation temperature. The values of critical and maximum elimination capacity ranged from 0.07 to 0.15 and from 0.13 to 0.16 g-C/h-kg packing material, respectively.

Acetone↗

Bacterial acetone and butanol production by industrial fermentation in the Soviet Union: use of hydrolyzed agricultural waste for biorefinery.

Clostridial acetone-butanol fermentation from renewable carbohydrates used to be the largest biotechnological process second only to yeast ethanol fermentation and the largest process ever run under sterile conditions. With the rising prices for mineral oil, it has now the economical and technological potential to replace petrochemistry for the production of fuels from renewable resources. Various methods for using non-food biomass such as cellulose and hemicellulose in agricultural products and wastes have been developed at laboratory scale. To our knowledge, the AB plants in Russia were the only full-scale industrial plants which used hydrolyzates of lignocellosic waste for butanol fermentation. These plants were further developed into the 1980s, and the process was finally run in a continual mode different from plants in Western countries. A biorefinery concept for the use of all by-products has been elaborated and was partially put into practice. The experience gained in the Soviet Union forms a promising basis for the development of modern large-scale processes to replace a considerable fraction of the current chemical production of fuel for our future needs on a sustainable basis.

Acetone↗

Utilisation of saccharides in extruded domestic organic waste by Clostridium acetobutylicum ATCC 824 for production of acetone, butanol and ethanol.

Domestic organic waste (DOW) collected in The Netherlands was analysed and used as substrate for acetone, butanol and ethanol (ABE) production. Two different samples of DOW, referred to as fresh DOW and dried DOW, were treated by extrusion in order to expand the polymer fibres present and to obtain a homogeneous mixture. The extruded material was analysed with respect to solvent and hot water extractives, uronic acids, lignin, sugars and ash. The total sugar content in the polymeric fractions of the materials varied from 27.7% to 39.3% (w/w), in which glucose represented the 18.4 and 25.1% of the materials, for fresh and dried DOW, respectively. The extruded fresh DOW was used as substrate for the ABE fermentation by the solventogenic strain Clostridium acetobutylicum ATCC 824. This strain was grown on a suspension of 10% (w/v) DOW in demineralised water without further nutrient supplement. This strain produced 4 g ABE/100 g extruded DOW. When C. acetobutylicum ATCC 824 was grown on a suspension of 10% (w/v) DOW hydrolysed by a combination of commercial cellulases and beta-glucosidases, the yield of solvents increased to 7.5 g ABE/100 g extruded DOW. The utilisation of sugar polymers in both hydrolysed and non-hydrolysed DOW was determined, showing that only a small proportion of the polymers had been consumed by the bacteria. These results indicate that growth and ABE production on DOW is mainly supported by soluble saccharides in the medium.

Acetone↗

Effects of methyl ethyl ketone, acetone, or toluene coadministration on 2,5-hexanedione concentration in the sciatic nerve, serum, and urine of rats.

OBJECTIVE: To clarify changes in the serum, nerve, and urinary levels of 2,5-hexanedione (2,5-HD) in rats on coadministration with methyl ethyl ketone (MEK), acetone (AC), and toluene (TO). METHOD: 2,5-HD alone or combined with MEK, AC, and TO was injected subcutaneously into a total of 306 male Wistar rats. The rats were divided as follows into 7 groups: (1) 2.6 mmol/kg 2,5-HD alone (HD) and (2) 2.6 mmol/ kg 2,5-HD combined with 2.6 mmol/kg MEK (HD + MEK), (3) with 2.6 mmol/kg AC (HD + AC), (4) with 2.6 mmol/kg TO (HD + TO), (5) with 13.0 mmol/kg MEK (HD + 5MEK), (6) with 13.0 mmol/kg AC (HD + 5AC), and (7) with 13.0 mmol/kg TO (HD + 5TO). 2,5-HD concentrations in the serum, sciatic nerve, and urine of rats were determined within 16 h of the injections and pharmacokinetic parameters were estimated. RESULTS: It was observed that (1) the 2,5-HD concentration and AUC value (area under concentration versus time curve) determined in the serum and nerve increased significantly in the cotreated groups as compared with the HD group; (2) the effect MEK had in elevating the 2,5-HD concentration and AUC in the serum and nerve was stronger than that of AC, and the effect AC had was stronger than that of TO; (3) a. dose increase from 2.6 to 13.0 mmol/kg for MEK and AC induced further increases in the 2,5-HD concentration and AUC determined in the serum and nerve; (4) elimination constants recorded for 2,5-HD (Ke) from the serum and nerve decreased in all the cotreated groups, and the degree of the decrease correlated inversely with the elevation in 2,5-HD concentration and AUC in the serum and nerve; and (5) urinary 2,5-HD concentrations measured in the 13.0-mmol/kg cotreated groups increased in parallel with the elevation in serum 2,5-HD concentrations. CONCLUSION: Coadministration of 2,5-HD with MEK, AC, or TO can increase the concentration and AUC of 2,5-HD in serum and the sciatic nerve, and these increases can be further enhanced by an increase in the concomitant doses of MEK and AC.

Acetone↗

Performance evaluation of acetone-butanol continuous flash extractive fermentation process.

A general mathematical model for performance evaluation of acetone-butanol continuous flash extractive fermentation system was formulated in the terms of productivity, energy requirement (energy utilization efficiency) and product purity. The simulation results based on experimental data showed that the most pronounced performance improvement could be achieved by using highly concentrated substrate as feed and the increase in solvent dilution rate could only improve the total productivity at the expense of energy utilization efficiency. A two-vessel partial flash system, with the first vessel of two to three plates and the second vessel as a complete flash vessel, is required to ensure high product purity. This method of mathematical formulation and analysis could be used and extended as a general tool to other bioprocesses equipped with multiple in situ recovery systems.

Acetone↗

Ab initio computational insight into the ion-pair S(N)2 reaction of lithium isothiocyanate and methyl fluoride in the gas phase and in acetone solution.

The ion-pair S(N)2 reaction LiNCS + CH3F with two mechanisms, inversion and retention, was investigated at the MP2(full)/6-311+G**//HF/6-311+G** level in the gas phase and in acetone solution. All HF-optimized structures were confirmed by vibrational frequency analysis. Based on IRC analyses, eight possible reaction pathways in the title reaction are proposed. The inversion mechanism through a six-membered-ring transition-state structure is the most favorable. Methyl thiocyanate should form preferentially in the gas phase and the more stable methyl isothiocyanate will be the main product in CH3COCH3. The retardation of the reaction in CH3COCH3 solution was attributed to the differences in the solvation free energies in the separated reactants and transition structures. All of the theoretical results are consistent with the experiment.

Acetone↗

Equilibrium yields of mono- and di-lauroyl mannoses through lipase-catalyzed condensation in acetone in the presence of molecular sieves.

Modification was made to our previously reported method to predict the equilibrium yields for the synthesis of mono- and di-lauroyl mannoses through the lipase-catalyzed condensation of lauric acid and mannose in acetone in the presence of molecular sieves. HPLC and mass spectra (MS) analyses indicated that two types of dilauroyl mannoses, which would be positional isomers of each other and are designated dilauroyl mannose I and II, were produced as well as monolauroyl mannose. The predicted yields of total mannose esters and dilauroyl mannose I agreed well with the experimental ones on the whole. The equilibrium yields of dilauroyl mannose II were higher than the predicted values, while the experimental values of monolauroyl mannose were lower than the predicted values.

Acetone↗

Consistent sets of spectrophotometric chlorophyll equations for acetone, methanol and ethanol solvents.

A set of equations for determining chlorophyll a (Chl a) and accessory chlorophylls b, c2, c1 + c2 and the special case of Acaryochloris marina, which uses Chl d as its primary photosynthetic pigment and also has Chl a, have been developed for 90% acetone, methanol and ethanol solvents. These equations for different solvents give chlorophyll assays that are consistent with each other. No algorithms for Chl c compounds (c2, c1 + c2) in the presence of Chl a have previously been published for methanol or ethanol. The limits of detection (and inherent error, +/- 95% confidence limit), for chlorophylls in all organisms tested, was generally less than 0.1 microg/ml. The Chl a and b algorithms for green algae and land plants have very small inherent errors (< 0.01 microg/ml). Chl a and d algorithms for Acaryochloris marina are consistent with each other, giving estimates of Chl d/a ratios which are consistent with previously published estimates using HPLC and a rarely used algorithm originally published for diethyl ether in 1955. The statistical error structure of chlorophyll algorithms is discussed. The relative error of measurements of chlorophylls increases hyperbolically in diluted chlorophyll extracts because the inherent errors of the chlorophyll algorithms are constants independent of the magnitude of absorbance readings. For safety reasons, efficient extraction of chlorophylls and the convenience of being able to use polystyrene cuvettes, the algorithms for ethanol are recommended for routine assays of chlorophylls. The methanol algorithms would be convenient for assays associated with HPLC work.

Acetone↗