Oxidation of catechol in plants : III. Purification and properties of the diphenylene dioxide 2,3-quinone-forming enzyme system from Tecoma leaves

In attempting to determine the nature of the enzyme system mediating the conversion of catechol to diphenylenedioxide 2,3-quinone, in Tecoma leaves, further purification of the enzyme was undertaken. The crude enzyme from Tecoma leaves was processed further by protamine sulfate precipitation, positive adsorption on tricalcium phosphate gel, and elution and chromatography on DEAE-Sephadex. This procedure yielded a 120-fold purified enzyme which stoichiometrically converted catechol to diphenylenedioxide 2,3-quinone. The purity of the enzyme system was assessed by polyacrylamide gel electrophoresis. The approximate molecular weight of the enzyme was assessed as 200,000 by gel filtration on Sephadex G-150. The enzyme functioned optimally at pH 7.1 and at 35 °C. The Km for catechol was determined as 4 × 10−4 Image . The enzyme did not oxidize o-dihydric phenols other than catechol and it did not exhibit any activity toward monohydric and trihydric phenols and flavonoids. Copper-chelating agents did not inhibit the enzyme activity. Copper could not be detected in the purified enzyme preparations. The purified enzyme was not affected by extensive dialysis against copper-complexing agents. It did not show any peroxidase activity and it was not inhibited by catalase. Hydrogen peroxide formation could not be detected during the catalytic reaction. The enzymatic conversion of catechol to diphenylenedioxide 2,3-quinone by the purified Tecoma leaf enzyme was suppressed by such reducing agents as GSH and cysteamine. The purified enzyme was not sensitive to carbon monoxide. It was not inhibited by thiol inhibitors. The Tecoma leaf was found to be localized in the soluble fraction of the cell. Treatment of the purified enzyme with acid, alkali, and urea led to the progressive denaturation of the enzyme.

Zeta Potential Studies On Some Iron-Ore Minerals In The Presence Of Starches

Authors perform zeta potential studies on hematite, corundum, and quartz samples using starches to understand the adsorption behavior of polymeric starch flocculants at the oxide mineral-solution interface and to correlate this information with their flocculation characteristics and investigate effects of pH and CaCl#72 on zeta potential of Fe ore minerals.

Flocculation, filtration and selective flocculation studies on haematite ore fines using starch

The flocculation and filtration characteristics of typical Indian iron ore fines have been studied using starch as flocculant in the presence of an inorganic electrolyte, namely calcium chloride. The effect of various parameters such as pH, starch and calcium chloride concentrations and pulp density on the settling and filtration rates, turbidity of the supernatant and on residual starch and calcium ion concentrates has been investigated through a statistical design and analysis approach and subsequently optimised on a laboratory scale. The adsorption mechanisms of starch onto haematite have been elucidated through adsorption density measurements, infrared and X-ray photoelectron spectroscopic techniques. The rheological property of the polymer solutions of relevance to flocculations has also been investigated. Further, the role of metal ion-starch interactions in the bulk solution, has been studied. In order to understand the nature of polymer adsorption at the double-layer, electrokinetic studies have been carried out with the iron ore mineral samples using starch and calcium chloride. Based on the above findings, selective floculaation tests on artificial mixtures of iron ore minerals have been carried out to determine the separation efficiencies from the view point of alumina and silica removal from haematite as well as the control of alumina: silica ratio in Indian iron ores.

Studies on nucleotidases in plants: Fluorescence and kinetic properties of nucleotide pyrophosphatase from mung bean (Phaseolus aureus) seedlings

Nucleotide pyrophosphatase of mung bean seedlings has earlier been isolated in our laboratory in a dimeric form (Mr 65,000) and has been shown to be converted to a tetramer by AMP and to a monomer by p-hydroxymercuribenzoate. All the molecular forms were enzymatically active with different kinetic properties. By a modified procedure using blue-Sepharose affinity chromatography, we have now obtained a dimeric form of the enzyme which is desensitized to AMP interaction. The molecular weight of the desensitized form of the enzyme was found to be the same as that of the native dimeric enzyme. However, the desensitized enzyme functioned with a linear time course, contrary to the biphasic time course exhibited by the native enzyme. In addition, it was not converted to a tetramer on the addition of AMP, had only one binding site for adenine nucleotides, and p-hydroxy-mercuribenzoate had no effect on the time course of the reaction or on the molecular weight of the enzyme. The temperature optimum of the desensitized enzyme was found to be 67 °C in contrast to the optimum of 49 °C for the native dimer. Fifty percent of the tryptophan residues of the desensitized enzyme were not accessible for quenching by iodide. Fluorescence studies gave Kd values of 0.34, 2.2, and 0.8 mImage for AMP, ADP, and ATP, which were close to the Ki values of 0.12, 2.2, and 0.9 mImage , respectively, for these nucleotides. The binding and inhibition studies with AMP and its analogs showed that the 6-amino group and the 5′-phosphate group were essential for the inhibition of the enzyme activity.

Studies in biogas technology. Part III. Thermal analysis of biogas plants

A thermal model for a conventional biogas plant has been developed in order to understand the heat transfer from the slurry and the gas holder to the surrounding earth and air respectively. The computations have been performed for two conditions : (i) when the slurry is at an ambient temperature of 20°C, and (ii) when it is at 35°C, the optimum temperature for anaerobic fermentation. Under both these conditions, the gas holder is the major “culprit” with regard to heat losses from the biogas plant. The calculations provide an estimate for the heat which has to be supplied by external means to compensate for the net heat losses which occur if the slurry is to be maintained at 35°C. Even if this external supply of heat is realised through (the calorific value of) biogas, there is a net increase in the biogas output, and therefore a net benefit, by operating the plant at 35°C. At this elevated temperature, the cooling effect of adding the influent at ambient temperature is not insignificant. In conclusion, the results of the thermal analysis are used to define a strategy for operating biogas plants at optimum temperatures, or at higher temperatures than the ambient.

Laboratory studies on ball wear in the grinding of a hematite-magnetite ore

Marked-ball grinding tests were carried out under different grinding conditions and environments. Three types of balls were used, namely, cast hyper steel, high chrome cast iron and EN-31 (forged), which cover a wide range of chemical composition, microstructure and media hardness. The effect of pulp density on ball wear and grinding efficiency was also studied. Relative pulp viscosities at different percent solids for the ore slurry were also determined. As the Kudremukh ore contained about 0.2% pyrite, the effect of addition of pyrite on ball wear was studied separately. Results of marked-ball grinding tests indicated that ball wear increased with time and showed a sharp increase for wet grinding over dry grinding. Ball wear under wet grinding conditions was also influenced by the gaseous atmosphere in the mill. At 70% solids, the best results in terms of reduced ball wear coupled with satisfactory grinding efficiency were obtained. The influence of oxygen on the corrosive wear of grinding balls was increasingly felt only if sulphide minerals such as pyrite were also present in the ore. The various ball materials could be arranged in the following order with respect to their overall wear resistance: high chrome cast iron > EN-31 (forged) > cast hyper steel.Possible ball wear mechanisms involved in the grinding of Kudremukh ore are discussed.

Ball wear and its control in the grinding of a lead-zinc sulphide ore

Marked ball grinding tests were carried out in the laboratory with a lead-zinc sulphide ore under different experimental conditions using high carbon low alloy steel (cast and forged) and high chrome cast iron balls. Relative ball wear as a function of grinding period and milling conditions was evaluated for the different types of ball materials. The role of corrosion and abrasion-erosion in the wear of grinding media is brought out. Methods to minimise ball wear through control of mill atmosphere and addition of reagents are discussed.

A Geostatistical Study pf Alumina-Content in Ores of An Indian Iron-Ore Deposit

Mining and blending operations in the high grade iron ore deposit under study are performed to optimize recovery with minimal alumina content while maintaining required levels of other chemical component and a proper mix of ore types. In the present work the regionalisation of alumina in the ores has been studied independently and its effects on global and local recoverable tonnage as well as on alternatives of mining operations have been evaluated. The global tonnage recovery curves for blocks (20m x 20m x 12m) obtained by simulation closely approximated the curves obtained theoretically using a change of support under the discretised gaussian model. Variations in block size up to 80m x 20m x 12m did not affect the recovery as the horizontal dimensions of the blocks are small in relation to the range of the variogram. A comparison of the local tonnage recovery curves obtained through multiple conditional simulations made with that obtained by the method of uniform conditioning of block grades on an estimate of panel 100m x 100m x 12m panel grade reveals comparable results only in panels which have been well conditioned and possesing an ensemble simulation mean close to the ordinary kriged value for the panel. Study of simple alternative sequence of mining on the conditionally simulated deposit shows that concentration of mining operations simultaneously on a single bench enhances the fluctuation in alumina values of ore mined.

Studies on molecular evolution and structural features of double-headed inhibitors of ?-amylase and trypsin in plants

Plant seeds usually have high concentrations of proteinase and amylase inhibitors. These inhibitors exhibit a wide range of specificity, stability and oligomeric structure. In this communication, we report analysis of sequences that show statistically significant similarity to the double-headed alpha-amylase/trypsin inhibitor of ragi (Eleusine coracana). Our aim is to understand their evolutionary and structural features. The 14 sequences of this family that are available in the SWISSPROT database form three evolutionarily distinct branches. The branches relate to enzyme specificities and also probably to the oligomeric state of the proteins and not to the botanical class of the plant from which the enzymes are derived. This suggests that the enzyme specificities of the inhibitors evolved before the divergence of commercially cultivated cereals. The inhibitor sequences have three regions that display periodicity in hydrophobicity. It is likely that this feature reflects extended secondary structure in these segments. One of the most variable regions of the polypeptide corresponds to a loop, which is most probably exposed in the native structure of the inhibitors and is responsible for the inhibitory property.

Immune responses to hemagglutinin-neuraminidase protein of peste des petits ruminants virus expressed in transgenic peanut plants in sheep

Peste des petits ruminants (PPR) is an acute, highly contagious disease of small ruminants caused by a morbillivirus, Peste des petits ruminants virus (PPRV). The disease is prevalent in equatorial Africa, the Middle East, and the Indian subcontinent. A live attenuated vaccine is in use in some of the countries and has been shown to provide protection for at least three years against PPR. However, the live attenuated vaccine is not robust in terms of thermotolerance. As a step towards development of a heat stable subunit vaccine, we have expressed a hemagglutinin-neuraminidase (HN) protein of PPRV in peanut plants (Arachis hypogea) in a biologically active form, possessing neuraminidase activity. Importantly. HN protein expressed in peanut plants retained its immunodominant epitopes in their natural conformation. The immunogenicity of the plant derived HN protein was analyzed in sheep upon oral immunization. Virus neutralizing antibody responses were elicited upon oral immunization of sheep in the absence of any mucosal adjuvant. In addition, anti-PPRV-HN specific cell-mediated immune responses were also detected in mucosally immunized sheep. (C) 2010 Elsevier B.V. All rights reserved.