Xanthan and gellan degradation by bacteria of activated sludge.

Owing to increasing amounts of xanthan and gellan in food, cosmetics and pharmaceuticals, as well as in some technical spheres, studies were carried out on the xanthan and gellan degrading bacteria present in activated sludge. The activated sludge used in the study was able to degrade both carbohydrates over 7 days, with levels of xanthan and gellan utilizing microbes estimated at 10(5) cells/g of dry sludge weight. Isolating key degrading bacteria revealed the important role of genus Paenibacillus in xanthan degradation and prosthecate bacterium Verrucomicrobium sp. GD, which was capable of gellan utilization. Further tests performed with both strains showed they were able to degrade other types of carbohydrate polymers, and that Verrucomicrobium sp. GD did not possess extracellular free gellan depolymerase.

Disposal of waste liquor from 2,5-dichlorodisulfanilic acid production.

Three alternative procedures were worked out for disposal of waste liquor from production of 2,5-dichlorodisultanilic acid [CAS No. 88-50-6]. This liquor is a hazardous liquid waste which, if discharged into a wastewater treatment plant without any treatment, inhibits processes of biological treatment. The first alternative utilizes stabilization/solidification technology by using fluidized-bed combustion ash. The second alternative consists in treating the liquor in a flow-through column packed with active carbon black as an adsorbent and discharging of the treated liquor, which already displays an acceptable chemical oxygen demand level after removal of organic pollutants by adsorption, into a wastewater treatment plant. The third alternative comprises of passing the waste liquor through the column with active carbon black and stabilization/solidification of the treated liquor by fluidized bed combustion ash. Efficiency of the liquor disposal in cases of stabilization/solidification was evaluated by determining dissolved organic carbon content, conductivity, pH, and ecotoxicity of test specimens' leachates. From results it follows that the proposed procedures represent efficient disposal of given liquid waste.

From no-confidence to nitric oxide acknowledgement: a story of bacterial nitric-oxide reductase.

The review briefly summarizes current knowledge of the bacterial nitric-oxide reductase (NOR). This membrane enzyme consists of two subunits, the smaller one contains haem C and the larger one two haems B and nonhaem iron. The protein sequence and structure of metal centres demonstrate the relationship of NOR to the family of terminal oxidases. The binuclear Fe-Fe reaction centre, consisting of antiferromagnetically coupled haem B and nonhaem iron, is analogous to Fe-Cu centre of terminal oxidases. The data on the structure and function of NOR and terminal oxidases suggest that all these enzymes are closely evolutionally related. The catalytic properties are determined most of all by the relatively high toxicity of nitric oxide as a substrate and the resulting strong need to maintain its concentration at nanomolar levels. A kinetic model of the action of the enzyme comprises substrate inhibition. NOR does not conserve the free energy of nitric oxide reduction because it does not work as a proton pump and, moreover, the protons coming into the reaction are taken from periplasm, i.e. they do not cross the membrane.

Kinetic analysis of substrate inhibition in nitric oxide reductase of Paracoccus denitrificans.

The current kinetic model for the nitric oxide reductase reaction (Girsch, P., and de Vries, S. (1997) Biochim. Biophys. Acta 1318, 202-216) does not involve the concentration of an electron donor. Here we introduce this variable and show, both theoretically and experimentally, its role in determining the extent of substrate inhibition by the excess of nitric oxide. NO is found to inhibit competitively with the electron donor, possibly by binding to the oxidized form of the enzyme. The observed partial character of the inhibition is tentatively explained by a slow reduction of the non-productive NO complex.

Pseudoazurin mediates periplasmic electron flow in a mutant strain of Paracoccus denitrificans lacking cytochrome c550.

A periplasmic protein able to transfer electrons from cytoplasmic membrane to the periplasmic nitrite reductase (cytochrome cd1) has been purified from the anoxically grown cytochrome c550 mutant strain Pd2121 and shown to be pseudoazurin by several independent criteria (molecular mass, copper content, visible spectrum, N-terminal amino acid sequence). Under our assay conditions, the half-saturation of electron transport occurred at about 10 microM pseudoazurin; the reaction was retarded by increasing ionic strength.

Evaluation of relative contributions of two enzymes supposed to metabolise hydrogen peroxide in Paracoccus denitrificans.

A biosensor exploiting an electrochemically mediated enzyme-catalysed reaction was used to quantify relative contributions of cytoplasmic catalase and periplasmic cytochrome c peroxidase to the overall rate of hydrogen peroxide breakdown in cells of Paracoccus denitrificans. The effects of antimycin (an inhibitor of electron flow to cytochrome c peroxidase), the reaction rate versus substrate concentration profiles for the whole cells and subcellular fractions, and the time courses of oxygen concentration demonstrated a profound decrease in the capacity of cytochrome c peroxidase to reduce H2O2 under in vivo conditions. The reason is suggested to be a competition for available electrons between the enzyme and terminal oxidases metabolising oxygen produced by catalase.