Thermostability and activation by divalent cations of the membrane-bound inorganic pyrophosphatase of Rhodospirillum rubrum

• 1.1. The activation energy of the membrane bound H⁺-pyrophosphatase is 44.9 k J·mol⁻¹, for the detergent solubilized enzyme is 55.9 kJ·mol⁻¹.
• 2.2. The Arrhenius plots obtained for pyrophosphatases of Rhodospirillum rubrum show no breaks.
• 3.3. At 70°C, the membrane-bound pyrophosphatase is more stable in the presence of either Mg²⁺ or Zn²⁺ than in their absence.
• 4.4. At 65°C, an activator effect of Mg²⁺ or Zn²⁺ was observed. Nevertheless, at 70°C no activation was obtained.
• 5.5. The activator effects of Mg²⁺ or Zn²⁺ were depended of their concentration.

     

Enzyme reactions in a multicompartment electrolyzer with isoelectrically trapped enzymes

The possibility of performing bioconversions under an electric field is here reported. A system is described by which the enzyme is trapped by an isoelectric mechanism between two zwitterionic membranes having pI values encompassing the isoelectric point of the enzyme. The enzyme is loaded into a multicompartment electrolyzer and kept operating under an electric field, which will continuously harvest the reaction product. Since, under focusing conditions, all buffering ions will vacate the reaction chamber at steady state, the buffering ion is trapped into the enzyme chamber by using amphoteric buffers co-isoelectric with the enzyme. As an example of such ‘isoelectrically immobilized’ reactor, the enzyme β-hydroxysteroid dehydrogenase is blocked into an isoelectric trap delimited by a pI 8.0 and a pI 6.5 membranes. 100 mM histidine (pI 7.47) is co-immobilized by the same isoelectric mechanism into the enzyme chamber. The dehydrocholic acid substrate (3,7,12-trioxo-5β-cholanoic acid) and reduced co-factor (NADH) are continuously infused into the enzyme chamber and the product (3β-hydroxy-7,12-dioxo-5β-cholanoic acid, a compound of pharmaceutical interest) and the oxidized co-factor (NAD⁺) collected, separately, into the two neighbouring chambers at the anodic side. Advantages: in a soluble form, the enzyme maintains the reaction kinetics of the free soluble form. Additionally, the reaction product and exhausted co-factor can be recovered by electrophoretic transport.     

Recovery and removal of uranium by using plant wastes

The uranium-adsorbing abilities of seven plant wastes were investigated. High abilities to adsorb uranium from non-saline water containing 10 mg dm⁻³ of uranium were observed with a number of plant wastes tested. However, with seawater supplemented with 10 mg dm⁻³ of uranium, similar results were found only with chestnut residues. When the plant wastes were immobilized with formaldehyde, their ability to adsorb uranium was increased. Uranium and copper ions were more readily adsorbed by all plant wastes tested than other metal ions from a solution containing a mixture of seven different heavy metals. The selective adsorption of heavy metal ions differs with different species of plant wastes. The immobilization of peanut inner skin, orange peel and grapefruit peel increased the selectivity for uranium.     

Anaerobic treatment of baker's yeast wastewater: I. Start-up and sodium molybdate addition

The anaerobic treatment of baker's yeast wastewater was studied using an anaerobic biological contact reactor (AnRBC) and a fixed-film reactor. The AnRBC had an active biomass developed within the reactor before this study commenced; however, the fixed-film reactor was started without attached biomass in a support structure. The gas production rates obtained for the AnRBC were between 0·55 and 0·61 litre methane per litre reactor per day. However, a gas production rate of only 0·46 litre methane per litre reactor per day was achieved after a four-month operating period for the fixed-film reactor. Higher chemical oxygen demand reduction was also found in the AnRBC. The results indicated that the presence of high sulfate concentration in baker's yeast wastewater affected teh start-up process. The reactor with fully developed active biomass was less susceptible to sulfate inhibition and showed improved anaerobic digestion. Results indicate that the reactor should be innoculated by feeding nutrient-balanced substrate before it was subjected to the digestion of baker's yeast wastewater. The fixed-film reactor was also fed with the substrate contianing sodium molybdate, an inhibitor of sulfate-reducing bacteria. The results indicated that both methanogenic and sulfate-reducing bacteria were inhibited.     

Enhancing H2O2 resistance of an esterase from Pyrobaculum calidifontis by structure-guided engineering of the substrate binding site

Applied Microbiology and Biotechnology - Green technologies are attracting increasing attention in industrial chemistry where enzymatic reactions can replace dangerous and environmentally...     

Sirtuin 5: a review of structure,known inhibitors and clues for developing new inhibitors

Sirtuins (SIRTs) are nicotinamide adenine dinucleotide (NAD+)-dependent protein deacetylases, which regulate important biological processes ranging from apoptosis, age-associated pathophysiologies, adipocyte and muscle differentiation, and energy expenditure to gluconeogenesis. Very recently, sirtuin 5 (SIRT5) has received considerable attention due to that it was found to have weak deacetylase activity but strong desuccinylase, demalonylase and deglutarylase activities, and it was also found to be associated with several human diseases such as cancer, Alzheimer’s disease, and Parkinson’s disease. In this review, we for the first time summarized the structure characteristics, known peptide and smallmolecule inhibitors of SIRT5, extracted some clues from current available information and introduced some feasible, practical in silico methods, which might be useful in further efforts to develop new SIRT5 inhibitors.     

Two new pyrrole derivatives isolated from Salacia amplifolia Merr

Two new pyrrole derivatives Salaciamole (1), Salaciaglycoside A (2), long with one known compound 1H-pyrrole-3-carboxylic acid (3) were isolated from the roots of Salacia amplifolia Merr (Hippocrateaceae). The structures of the new compounds were deduced from their comprehensive spectroscopic analysis including IR, HR-EI-MS, ¹H NMR, ¹³C NMR, DEPT, COSY, HMBC and HMQC. And the structure of the known compound was identified by comparison of their spectral data with those reported in the literature.     

Developmental regulation of phospholipase D in tomato fruits

The catabolism of phospholipids initiated by phospholipase D (PLD, EC 3.1.4.4) is an inherent feature of developmental processes that include fruit growth and ripening. In cherry tomatoes (Lycopersicon esculentum Mill.), soluble and membrane-associated PLD activities increased during fruit development, which peaked at the mature green and orange stages. The increase in PLD activity was associated with a similar increase in the intensity of a 92 kDa band as demonstrated by western blot analysis. A full-length cDNA having 2430 bp and encoding a putative polypeptide with 809 amino acids, was isolated using tomato RNA, RT-PCR and 5' and 3' rapid amplification of cloned ends (RACE). Analysis of the primary and secondary structures showed the presence of the C2 domain, the PLD domain and several other features characteristic of PLD alpha. Microtom tomato plants transformed with antisense PLD alpha cDNA, were similar to untransformed plants and showed normal fruit set and development. The ethylene climacteric was delayed by over 7 d in the antisense PLD fruits, indicative of a slower ripening process. The leaves and unripened fruits of antisense PLD microtom plants possessed lowered PLD activity and PLD protein, as demonstrated by western blotting. However, during ripening, PLD activity in the transgenic fruits was maintained at a higher level than that in the untransformed control. Immunolocalization of PLD in microtom tomato fruits revealed the cytosol-membrane translocation of PLD during fruit development. The ripe fruits of antisense PLD celebrity plants possessed lowered PLD expression and activity and showed increased firmness and red colour. These results suggest that the expression of antisense PLD cDNA could be variable in different tomato varieties. The potential role of PLD in ethylene signal transduction events is discussed.