Energy production in Gluconobacter liquefaciens.
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L-Ascorbic acid is an important product currently made using the Reichstein process, which is mainly chemical. Recently, bacteria have been identified that are able to transform in a very efficient way glucose to 2,5-keto-D-gluconic acid and this product to 2-keto-L-idonic acid, precursor of L-ascorbic acid. When the corresponding strains are used together, it is possible to get 2-keto-L-idonic acid directly from glucose. Moreover, new strains have been constructed by introducing a gene from a strain responsible for the second step into a strain responsible for the first step. By using one of the new strains, the transformation can be performed in a single step with only one strain. However, the classical process still remains the most competitive.
Arsenic contamination in ground water poses a serious threat on human health. The tea fungus, a waste produced during black tea fermentation has been examined for its capacity to sequester the metal ions from ground water samples. Autoclaved tea fungal mat and autoclaving followed by FeCl3 pretreated tea fungal mat were exploited for removal of As(III), As(V) and Fe(II) from ground water sample collected from Kolkata, West Bengal, India. The biosorption rate tends to increase with the increase in contact time and adsorbent dosage. FeCl3 pretreated and autoclaved fungal mats removed 100% of As(III) and Fe(II) after 30 min contact time and 77% of As(V) after 90 min contact time. The optimum adsorbent dosage was 1.0 g/50 mL of water sample. The results revealed that the FeCl3 pretreated fungal mat could be used as an effective biosorbent for As(III) and As(V); autoclaved fungal mat for Fe(II) removal from ground water sample.
Kombucha is a traditional fermentation of sweetened tea, involving a symbiosis of yeast species and acetic acid bacteria. Despite reports of different yeast species being associated with the fermentation, little is known of the quantitative ecology of yeasts in Kombucha. Using oxytetracycline-supplemented malt extract agar, yeasts were isolated from four commercially available Kombucha products and identified using conventional biochemical and physiological tests. During the fermentation of each of the four products, yeasts were enumerated from both the cellulosic pellicle and liquor of the Kombucha. The number and diversity of species varied between products, but included Brettanomyces bruxellensis, Candida stellata, Schizosaccharomyces pombe, Torulaspora delbrueckii and Zygosaccharomyces bailii. While these yeast species are known to occur in Kombucha, the enumeration of each species present throughout fermentation of each of the four Kombucha cultures demonstrated for the first time the dynamic nature of the yeast ecology. Kombucha fermentation is, in general, initiated by osmotolerant species, succeeded and ultimately dominated by acid-tolerant species.
l-Ribulose is an important chiral lead molecule used for the synthesis of, among others, l-ribose, a high-value rare sugar used in the preparation of antiviral drugs. These drugs--nucleoside-analogues--gain importance in the treatment of severe viral diseases, like those caused by the HIV or hepatitis virus. In this study, factors that may have an impact on l-ribulose production with Gluconobacter oxydans and on the stability of l-ribulose were investigated. A bioconversion-type process, using washed resting cells, was chosen to produce l-ribulose from ribitol. In this process, the cell production and bioconversion phase were separated. The former was first optimized and a maximum cell mass of 1.5 g CDWL(-1) could be produced. For the bioconversion phase, the aeration level of the system proved to be one of the most critical factors; a maximal production rate of 15.7 g L(-1)h(-1) or 5.9 g(g CDW)(-1)h(-1) of l-ribulose could be reached. Furthermore, resting cells were found capable of completely converting ribitol solutions of up to 300 g L(-1) within 30 h, although the kinetics indicated a rather low affinity of the dehydrogenase enzymes for the substrate.
A method has been developed that has been found to give reproducible images of uncoated polysaccharides by Atomic Force Microscopy (AFM). Aqueous solutions of the polysaccharide are deposited as drops onto freshly cleaved mica surfaces, air dried, and then imaged under butanol. The method has been used to obtain images of the bacterial polysaccharide acetan. In regions within the deposited sample, where the molecules are aligned side-by-side, it has been possible to observe a periodic structure along the polysaccharide chain, attributable to the helical structure of acetan.
It has been discovered that deacetylation of the bacterial polysaccharide acetan promotes synergistic interactions with either locust bean gum (LBG) or konjac mannan (KM). Acetan is similar in structure to xanthan, and adopts a similar 5-fold conformation in the solid state. Like xanthan, it shows a thermally reversible order (helix)-disorder (coil) transition in solution. Both polymers have a cellulosic backbone with charged (anionic) sidechains attached at O-3 of alternate glucosyl residues, but the sidechains in acetan are longer (pentasaccharide rather than trisaccharide) and do not contain pyruvic substituents. Acetan has two sites of acetylation, one at O-6 of the inner mannosyl residue of the carbohydrate sidechains (as in xanthan) and the other on the polymer backbone (believed to be at O-6 of the branched glucosyl residues). Solutions of acetan or deacetylated acetan were equilibrated against 10 mM potassium chloride (to stabilise the ordered conformation) and were mixed (at 25 degrees C) with solutions of LBG or KM, also equilibrated against 10 mM potassium chloride. Unlike xanthan, native acetan showed no evidence of synergistic interaction with either LBG or KM. After deacetylation, however, large enhancements were observed in dilute-solution viscosity, and thermoreversible gels were formed at higher concentrations. With KM as co-synergist, gel melting was accompanied by an intense endotherm in differential scanning calorimetry. The magnitude of this endotherm increased with storage time at 25 degrees C, reaching a final value of delta H approximately 15.9 J/g (in comparison with delta H approximately 5.0 J/g for the order-disorder transition of deacetylated acetan alone). It is suggested that interaction occurs by formation of heterotypic junctions between the acetan backbone and unsubstituted regions of the plant polysaccharide, and that the acetate groups on native acetan promote solubility and hence inhibit association.
Binary mixtures of deacetylated acetan and konjac mannan form thermoreversible gels under conditions for which the individual components do not gel. Such synergistic behaviour is normally attributed to intermolecular binding between the two polysaccharides. X-ray diffraction data obtained from oriented fibres prepared from deacetylated acetan-konjac mannan gels provides direct evidence for intermolecular binding between the two polysaccharides. The novel heterotypic junction zones appear to be six-fold helices with a pitch of 5.6 +/- 0.1 nm.
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Detailed characterization of enzyme susceptibility of bacterial cellulose containing N-acetylglucosamine (GlcNAc) residues (N-AcGBC) which possess high susceptibility for cellulase and lysozyme and slight susceptibility for chitinase was studied. Turbidimetric lysozyme assay of N-AcGBC showed that (i) the susceptibilities of various N-AcGBCs for lysozyme were proportional to GlcNAc content, and (ii) N-AcGBC homogenates were divided into two groups based on the rate of turbidity reduction (not dependent on GlcNAc content). High reactivity of N-AcGBC for lysozyme would arise from fine microfibrils characteristic of bacterial cellulose (BC) and random distribution of GlcNAc residues in N-AcGBC because water soluble oligomers of N-AcGBC produced by lysozymic hydrolysis did not inhibit lysozyme activity; however, the random distribution of GlcNAc seemed to result in the slight susceptibility of N-AcGBC for chitinase. The rate of cellulolytic turbidity reduction of N-AcGBC was slower than that of BC, which arose from the inhibition for binding of cellulase by GlcNAc residues.
Methods used in the manufacture of both ethanol and ascorbic acid have been developed over the past 150 years. The early stages of the development of both processes were characterized by the interaction between chemists, biologists and engineers, but social and economic influences also played their part. The history of these two examples illustrates the diversity of skills and influences needed for the application of biological catalysis to large-scale chemical manufacture.