Concept: PH indicator
The comparison of volumes of cells and subcellular structures with the pH values reported for them leads to a conflict with the definition of the pH scale. The pH scale is based on the ionic product of water, K(w) = [H(+)]×[OH(-)].We used K(w) [in a reversed way] to calculate the number of undissociated H(2)O molecules required by this equilibrium constant to yield at least one of its daughter ions, H(+) or OH(-) at a given pH. In this way we obtained a formula that relates pH to the minimal volume V(pH) required to provide a physical meaning to K(w), [Formula: see text] (where N(A) is Avogadro’s number). For example, at pH 7 (neutral at 25°C) V(pH) = 16.6 aL. Any deviation from neutral pH results in a larger V(pH) value. Our results indicate that many subcellular structures, including coated vesicles and lysosomes, are too small to contain free H(+) ions at equilibrium, thus the definition of pH based on K(w) is no longer valid. Larger subcellular structures, such as mitochondria, apparently contain only a few free H(+) ions. These results indicate that pH fails to describe intracellular conditions, and that water appears to be dissociated too weakly to provide free H(+) ions as a general source for biochemical reactions. Consequences of this finding are discussed.
Feasibility of soil washing to remediate Hg contaminated soil was studied. Dry sieving was performed to evaluate Hg distribution in soil particle size fractions. The influence of dissolved organic matter and chlorides on Hg dissolution was assessed by batch leaching tests. Mercury mobilization in the pH range of 3-11 was studied by pH-static titration. Results showed infeasibility of physical separation via dry sieving, as the least contaminated fraction exceeded the Swedish generic guideline value for Hg in soils. Soluble Hg did not correlate with dissolved organic carbon in the water leachate. The highest Hg dissolution was achieved at pH 5 and 11, reaching up to 0.3% of the total Hg. The pH adjustment was therefore not sufficient for the Hg removal to acceptable levels. Chlorides did not facilitate Hg mobilization under acidic pH either. Mercury was firmly bound in the studied soil thus soil washing might be insufficient method to treat the studied soil.
The characterization of a spore laccase from Bacillus vallismortis fmb-103, isolated from textile industry disposal sites, is described. The activity was 6.5U/g of dry spore with ABTS as the substrate. The enzyme was quite stable at high temperature. It retained more than 90% of its initial activity after 10h at 70°C. The enzyme demonstrated broad pH stability in both acidic and alkaline conditions. There was almost no activity loss at pH 3 over an extended period of time, and the relative activity remained at 82% and 38% at pH 7 and pH 9 after 10days. NaN(3), SDS, l-cysterine, Dithiothreitol, EDTA and NaCl inhibit the enzyme activity. Triphenylmethane dyes, including malachite green, brilliant green and aniline blue were efficiently degraded by the enzyme after 24h in combination with a mediator with efficiencies of 76.84%, 96.56% and 81.17%, respectively. The reusability of spore laccase for decolorization dyes was also examined.
An efficient and reversible ionic gating that can be activated by pH and light is demonstrated on page 6193 by Lei Jiang, Zhishan Bo, and co-workers, by modifying a malachite green derivative on the interior surface of an ion track-etched conical nanochannel. The switches between the OFF-state and the ON-state are dependent on the surface charge transition caused by the malachite green derivative, making it suitable for confined spaces. Such a dual-driven ionic gating could find applications in electronics, actuators, and biosensors.
A new type of pH-sensitive liposomes (fliposomes) was designed based on the amphiphiles that are able to perform a pH-triggered conformational flip (flipids). This flip disrupts the liposome membrane and causes rapid release of the liposome cargo, specifically in response to lowered pH. The flipids (1) and (2) are equipped with a trans-2-aminocyclohexanol conformational switch. pH-sensitive fliposomes containing one or both of these flipids, as well as POPC and PEG ceramide, were constructed and characterized. These compositions were stable at 4(°)C and pH 7.4 for several months. Fliposomes loaded with ANTS/DPX performed an unusually quick content release within a few seconds at pH below 8.5 (in case of 2) and 6.0 (in case of 1). This difference in pH sensitivity demonstrates a potential for the custom design of flipids by variation of the amino group to target areas with specific pH values. The pH titration curves for the fliposome leakage parallel the curves for the acid-induced conformational flip of 1 and 2 studied by (1)H NMR. A plausible mechanism of pH sensitivity starts with an acid-triggered conformational flip of 1 or 2, which changes the molecular size and shape, shortens the lipid tails, and perturbs the liposome membrane, resulting in the content leakage.
Research was conducted to determine suitable chemical parameters as indicators of odor from decomposing food wastes. Prepared food scraps were stored in 18l plastic buckets (2kg wet weight each) at 20°C and 8°C to reproduce high and low temperature conditions. After 1, 3, 7, 10 and 14days of storage, the odor from the buckets were marked to an intensity scale of 0 (no odor) to 5 (intense) and the corresponding leachate analyzed for volatile fatty acids, ammonia and total organic carbon. A linear relationship between odor intensity and the measured parameter indicates a suitable odor indicator. Odor intensified with longer storage period and warmer surroundings. The study found ammonia and isovaleric acid to be promising odor indicators. For this food waste mixture, offensive odors were emitted if the ammonia and isovaleric acid contents exceeded 360mg/l and 940mg/l, respectively.
A new sample preparation procedure, termed pH-controlled dispersive liquid-liquid microextraction (pH-DLLME), has been developed for the analysis of ionisable compounds in highly complex matrices. This DLLME mode, intended to improve the selectivity and to expand the application range of DLLME, is based on two successive DLLMEs conducted at opposite pH values. pH-DLLME was applied to determination of ochratoxin A (OTA) in cereals. The hydrophobic matrix interferences in the raw methanol extract (disperser, 1mL) were removed by a first DLLME (I DLLME) performed at pH 8 to reduce the solubility of OTA in the extractant (CCl(4), 400μL). The pH of the aqueous phase was then adjusted to 2, and the analyte was extracted and concentrated by a second DLLME (extractant, 150μL C(2)H(4)Br(2)). The main factors influencing the efficiency of pH-DLLME including type and volume of I DLLME extractant, as well as the parameters affecting the OTA extraction by II DLLME, were studied in detail. Under optimum conditions, the method has detection and quantification limits of 0.019 and 0.062μgkg(-1), respectively, with OTA recoveries in the range of 81.2-90.1% (n=3). The accuracy of the analytical procedure, evaluated with a reference material (cereal naturally contaminated with OTA), is acceptable (accuracy of 85.6%±1.7, n=5). The applicability of pH-DLLME to the selective extraction of other ionisable compounds, such as acidic and basic pharmaceutical products was also demonstrated. The additional advantages of pH-DLLME are a higher selectivity and the extension of this microextraction technique to highly complex matrices.
Production of the lignocellulose-degrading enzymes endo-1,4-β-glucanase, 1,4-β-glucosidase, cellobiohydrolase, endo-1,4-β-xylanase, 1,4-β-xylosidase, Mn peroxidase, and laccase was characterized in a common wood-rotting fungus Fomes fomentarius, a species able to efficiently decompose dead wood, and compared to the production in eight other fungal species. The main aim of this study was to characterize the 1,4-β-glucosidase produced by F. fomentarius that was produced in high quantities in liquid stationary culture (25.9 U ml(-1)), at least threefold compared to other saprotrophic basidiomycetes, such as Rhodocollybia butyracea, Hypholoma fasciculare, Irpex lacteus, Fomitopsis pinicola, Pleurotus ostreatus, Piptoporus betulinus, and Gymnopus sp. (between 0.7 and 7.9 U ml(-1)). The 1,4-β-glucosidase enzyme was purified to electrophoretic homogeneity by both anion-exchange and size-exclusion chromatography. A single 1,4-β-glucosidase was found to have an apparent molecular mass of 58 kDa and a pI of 6.7. The enzyme exhibited high thermotolerance with an optimum temperature of 60 °C. Maximal activity was found in the pH range of 4.5-5.0, and K (M) and V (max) values were 62 μM and 15.8 μmol min(-1) l(-1), respectively, when p-nitrophenylglucoside was used as a substrate. The enzyme was competitively inhibited by glucose with a K (i) of 3.37 mM. The enzyme also acted on p-nitrophenylxyloside, p-nitrophenylcellobioside, p-nitrophenylgalactoside, and p-nitrophenylmannoside with optimal pH values of 6.0, 3.5, 5.0, and 4.0-6.0, respectively. The combination of relatively low molecular mass and low K (M) value make the 1,4-β-glucosidase a promising enzyme for biotechnological applications.
A multi-responsive sensor was constructed by combining a ferrocene unit and a rhodamine block via a carbohydrazone bond. The sensor showed high selectivity toward Cu(2+) over other common metal ions in a wide pH range with excellent reversibility and rapid response. The obvious color change from colorless to pink upon the addition of Cu(2+) could make it a suitable ‘naked-eye’ indicator for Cu(2+). The detection limit (LOD) obtained was down to 2.0 nM and the association constant (Ka) was evaluated as 4.65 × 10(7) M(-1). The accuracy for detecting Cu(2+) in environmental river water was compared favorably with the traditional atomic absorption spectroscopy method (AAS). Finally, we proposed a reversible ring-opening mechanism (Off-On) of the rhodamine spirolactam induced by Cu(2+) binding and a 2 : 1 stoichiometric structure between and Cu(2+).
This pHLIP is no flop: Functionalizing mesoporous silica nanoparticles (MSNs) with pHLIPss peptide provides a controlled-release nanoparticle drug delivery system targeting the acidic tumor microenvironment. At low pH values, pHLIPss inserts into the cell membrane and translocates carriers into cells, where the cargo is released by the cleavage of the pHLIPss disulfide bonds.