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The acetone-butanol-ethanol fermentation.

Renewed interest in the acetone-butanol-ethanol (ABE) fermentation as a route for industrial production of butanol has been evident since the oil crisis of the 1970s. The present review includes an historical recap of the traditional industrial process and culturing practices useful in maintaining viable solvent-producing cultures, and then summarizes new and exciting research on the physiology and genetics of the microorganisms as well as process design. Most of these reports relate to improvements in solvent yield and the overall process, since traditional production is not efficient under present economic conditions. Conclusions are then made on future developments necessary for the establishment of an economically viable industrial process.

Acetone↗

Bacteriophage infections in the industrial acetone butanol (AB) fermentation process.

The reported incidence and effects of bacteriophage infections occurring in the industrial acetone butanol (AB) fermentation processes operated in the USA, Japan, and Puerto Rico during the earlier part of the twentieth century is reviewed. The growth characteristics and solvent-producing ability of a lysogenic strain of Clostridium madisonii isolated from a phage infection in Puerto Rico was determined in molasses fermentation medium. The host strain harbours a large lysogenic phage belonging to the Siphoviridae and the growth rate of the lysogenic strain was found to be slower than the non-lysogenic parent strain and exhibited reduced solvent production. The history of phage infections that occurred in the South African AB process is documented along with the various remedial actions that were taken to restore production. A more detailed account of the last phage infection that occurred in 1980 involving a small pseudo-lysogenic phage belonging to the Podoviridae is given. This phage infected Clostridium beijerinckii P260 and a number of closely related industrial strains. Factory-scale fermentations contaminated by this phage were compared with equivalent laboratory-scale control fermentations. The effect of the phage infection in the full-scale and laboratory-scale fermentations were monitored. Results obtained in laboratory-based studies included an assessment of the effect of the multiplicity of infection and the timing of phage infection. The general effects and symptoms of phage infections in the industrial AB fermentation are reviewed including gross changes in the fermentation and changes in cell morphology. Common techniques used for the diagnosis of phage infections and approaches for controlling phage contamination in the AB fermentation are discussed. Prevention strategies included good factory hygiene, sterilisation, decontamination and disinfection, and the use of resistant strains immunised against specific phages.

Acetone↗

The economics of acetone-butanol fermentation: theoretical and market considerations.

Acetone-butanol (AB) fermentation was once run commercially in many countries until these chemicals could be made more cheaply from fossil oil sources. Research into the revitalisation of the process has shown that the process could once again be run economically in niche markets if run in a relatively small industrial scale processing low-grade agricultural products. The following analysis is intended to help identify suitable niche markets.

Acetone↗

Conversion of solvent evaporation residues from the AB- (acetone - butanol) bioprocess into bacterial cells accumulating thermoplastic polyesters.

In a bioconversion study based on utilisation of by-products from the AB- (acetone - butanol) bioprocess a new isolated gram-negative solvent tolerant bacterium was used to convert the AB process residue after removal of the major part of the solvents. The bacterium identified as a representative of the genus Alcaligenes (designated as Alcaligenes sp. G) was capable of growth up to optical densities ranging from 8 to 20 and simultaneously of polyhydroxyalkanoate-(PHA-)accumulation up to 40% per dry weight. A standardised medium based on AB by-products containing 7 g/l of butyrate and 5 g/l of acetate at pH 7.5 was used in our studies for bioconversion into PHAs. Concentrations of 1-butanol, which is known for its membrane damaging properties in microorganisms, were tolerated in the AB by-products medium up to 4 g/l without significant inhibition of cellular growth. No inhibition of growth was observed, when the medium was adjusted to 40 g/l butyrate. Due to the toxicity of the remaining 1-butanol maintenance of sterility is of no high priority during the process. The use of acetate and butyrate from an AB process is expected to provide a higher return-on-investment than the combustion of biogas to help meet energy demands.

Acetone↗

The cause of "acid-crash" and "acidogenic fermentations" during the batch acetone-butanol-ethanol (ABE-) fermentation process.

Experiments were performed to determine the cause of "acid crash", a phenomenon which occasionally occurs in pH-uncontrolled batch fermentations resulting in premature cessation of ABE (acetone butanol) production. The results indicate that "acid crash" occurs when the concentration of undissociated acids in the broth exceeds 57 - 60 mmol/l. Prevention can be achieved by introducing some limited pH control to minimize the concentration of undissociated acids or by slowing the metabolic rate, and thus the rate of acid production, by, for example, lowering the fermentation temperature. "Acidogenic fermentations", which occur when batch fermentations are performed at pH values close to neutrality, are due to rapid production of acids followed by inhibition of solventogenesis when the total acid concentration reaches 240 - 250 mmol/l. Solventogenesis can be achieved at these pH values by lowering the glucose uptake rate / acid production rate by use of e.g. elevated glucose or lowered yeast extract concentrations in the growth medium.

Acetone↗

Conversion of solvent evaporation residues from the AB- (acetone-butanol) bioprocess into bacterial cells accumulating thermoplastic polyesters.

In a bioconversion study based on utilisation of by-products from the AB- (acetone-butanol) bioprocess a new isolated gram-negative solvent tolerant bacterium was used to convert the AB process residue after removal of the major part of the solvents. The bacterium identified as a representative of the genus Alcaligenes (designated as Alcaligenes sp. G) was capable of growth up to optical densities ranging from 8 to 20 and simultaneously of polyhydroxyalkanoate- (PHA-)accumulation up to 40% per dry weight. A standardised medium based on AB by-products containing 7 g/l of butyrate and 5 g/l of acetate at pH 7.5 was used in our studies for bioconversion into PHAs. Concentrations of 1-butanol, which is known for its membrane damaging properties in micro-organisms, were tolerated in the AB by-products medium up to 4 g/l without significant inhibition of cellular growth. No inhibition of growth was observed, when the medium was adjusted to 40 g/l butyrate. Due to the toxicity of the remaining 1-butanol maintenance of sterility is of no high priority during the process. The use of acetate and butyrate from an AB process is expected to provide a higher return-on-investment than the combustion of biogas to help meet energy demands.

1-Butanol↗

Photocatalytic degradation of dye pollutants in the presence of acetone.

The photodegradation of textile azo dyes in aqueous solution containing acetone (ACE) photosensitizer was investigated at 253.7 nm. In this study, a typical azo reactive dye (C I Reactive Red 2-RR2) is used as the probe compound. ACE is a powerful photosensitizer that possesses high triplet energy, and the presence of ACE significantly enhances the overall photodegradation reaction through simultaneous sensitization. In the UV/ACE system, at the optimum conditions, over 97% of RR2 can be successfully decomposed and decolored within 300 seconds of retention time. The photodegradation mechanisms of RR2 involve both photodechlorination and photoreduction, which follow pseudo first-order decay kinetics. The quantum yield of the photolysis system has been evaluated as a function of ACE concentration and the initial pH. RR2 contains the chlorotriazinyl group, which undergoes hydrolysis and results in the formation of hydrochloric acid, indicating that the photodechlorination process is the dominant reaction mechanism. The photo-product, H+, is likely to retard the quantum yield of dye decay, suggesting that the initial pH is a critical factor in changing the photodecolorization rates of dye. Generally, this system is more favorable at higher pH levels.

Acetone↗

Specificity and reproducibility of acetone precipitation in identifying surface-active phosphatidylcholine in amniotic fluid.

We show the thoretical and actual variability in the amount of surface-active lecithin precipitated in the acetone-precipitation step of determining lecithin/sphingomyelin (L/S) ratios in the clinical laboratory. Lecithins precipitated may range from 60 to 90% of the total surface-active lecithins present. The range is sufficient to account for the occasional "immature" L/S ratios (less than 2/1) found in amniotic fluid from women bearing mature fetuses.

Acetone↗

The effects of water and acetone-based dentin adhesives on apical microleakage.

In this study, the aim was to assess the in vitro apical microleakage of a resin-based sealer used with two different adhesives. Thirty nine freshly extracted maxillary incisors were used. The teeth were decoronated at the cemento-enamel junction with a water-cooled fissure bur. Chemo-mechanical debridement of the root canals was accomplished with the step-back technique. The smear layer was removed by 19% ethylenediamine tetra acetic acid (EDTA). The roots were then divided into three experimental groups of thirteen teeth in each. Specimens in group 1 were filled with gutta-percha, AH Plus sealer, and water-based adhesive system (Syntac Single Component). Group 2 specimens were filled with gutta-percha, AH Plus sealer, and acetone-based dentin adhesive (Prime & Bond NT ). Specimens of group 3 were filled with only gutta-percha and AH Plus sealer (no adhesive was applied). The teeth were immersed into 2% methylene blue solution. Apical sealing qualities were assessed by measuring the linear dye penetration with a stereomicroscope. Dentin tubule penetration was observed under scanning electron microscopy (SEM). Results showed no statistically significant difference between the materials used, however, the leakage in group 2 was less than group 1 and 3.

Acetone↗

Application of alcohol- or acetone-fixed Schistosoma eggs as an alternative to lyophilized eggs for the circumoval precipitin test.

Antigenicity of Schistosoma mansoni and S. japonicum eggs preserved in ethanol or acetone were assessed in a circumoval precipitin (COP) assay. The egg antigens were found to retain sufficiently their COP reactivity for the diagnosis of both schistosomiasis mansoni and japonica, although their reactivity became lower than that of lyophilized eggs. These alternative preparations for COP tests have advantages, such as keeping eggs directly in fixatives soon after the egg-purification process. Furthermore, evaporation-process may cause eggshell cleavages which facilitate the reaction. The possible usefulness of those eggs in COP assays in local endemic areas is discussed.

Acetone↗

[Structure of the reaction adduct of xanthothricin with acetone].

Reaction of xanthothricin demethylation by amines in acetone as a solvent yielded a new compound with a formula of C10H13N5O3. Mass spectrometry, 1H NMR-spectroscopy and x-ray analysis showed that the compound was 4H-1,6-dimethyl-4a(2-oxopropyl)-pyrimido-[5,4-e]-1 ,2,4-triazine-5,7-dione.

Acetone↗

Immunogenic complexes obtained from Salmonella typhi-murium and Salmonella typhi Ty2 by the bacterial acetone powder method.

An immunogenic complex was isolated from Salmonella typhi-murium and another one from Salmonella typhi Ty2. Both were prepared by the bacterial acetone powder method which eliminated the cell wall, the DNA almost completely and the membrane phospholipids. The complexes were denominated "New Vaccines". The S. typhi-murium new vaccine induced, even at doses of 0.5 microgram dry weight per mouse, a high degree of protection against the challenge of the virulent microorganism. By immunoelectrophoresis, 21 antigen-antibody systems could be detected, two of them corresponding to O antigens. The S. typhi Ty2 new vaccine induced better protection than the standard vaccine (heat-phenol inactivated typhoid vaccine) when both vaccines were compared in the relative potency test. Moreover, the new vaccine had very low toxicity when inoculated in humans at doses of 1 microgram dry weight, able to elicite a high antibody titre (1/1,790 mean of 10 sera) in 75% of the tested population, estimated by the complement fixation test. In contrast, the standard vaccine induced a low antibody titre (1/222, mean of 5 sera) in 50% of the humans inoculated with 1 X 10(8) bacterial cells. The new vaccine did not induce undesirable effects whereas the standard vaccine induced an important inflammatory process in 100% of the cases, with intense local pain in 67% after 24 h post-first inoculation as well as other less severe symptoms.

Acetone↗

Protein precipitation by acetone for the analysis of polyethylene glycol in intestinal perfusion fluid.

Canine ileal segments were luminally perfused with known amounts of polyethylene glycol (PEG), and samples obtained were freed of protein by a cold acetone (CA) method and assayed for PEG by trichloroacetic acid in the presence of calcium cations. This procedure was compared to the Barium salt (BaS) protein precipitation method of Hyden (Ann R Agric Coll Sweden 22:411-424, 1955). The CA method saved specimen volume and time in handling, whereas its stability, linearity, and reproducibility were equivalent to the more complex BaS method. In vitro studies disclosed no appreciable differences when PEG specimens were bile stained or dissolved in various solvents. A small modification of the CA method improved accuracy in measurement of small PEG concentrations.

Acetone↗

[Gas-chromatographic determination of methyl-isobutyl ketone, methyl-isobutyl carbinol, acetone, toluene and o-xylene vapors in the air].

A gas chromatographic method of determination of methyl isobutyl ketone, methyl isobutyl carbinol, acetone, toluene and o-xylene has been elaborated. Those compounds have been best separated on the column containing 10% Carbowax 20 M on Chromosorb W. Air samples were collected by passing the examined air through charcoal tubes and by consequent desorption of adsorbed compounds with carbon disulfide. Activated charcoal used in tests has been found to adsorb (by weight) 15% of the determined solvents.

Acetone↗