Adaptability in enzymes

In studying inhibitors and activators of enzymes it has often been found that inhibitors act as activators when in very low concentrations and that certain activators (in suitable concentrations) show inhibition. Similar results were obtained in the present work in studying urease from soya bean in the presence of lead acetate. Lead acetate is normally an inhibitor of urease. It was found, however, that in small concentrations of lead acetate increase in the activity of urease occurs. On prolonging the time of the reaction inhibition develops and still later increased enzymatic activity again occurs. The results were elaborated statistically and are discussed in relation to the use of “adaptation” of urease in unfavourable conditions in reaction mixtures by the action of lead acetate.     

Xylanolytic anaerobic thermophiles from Icelandic hot-springs

Anaerobic enrichment cultures inoculated with neutral and alkaline (pH 7.0–9.0) sediment and biomat samples from hot-springs in Hveragerdi and Fluir, Iceland, were screened for growth on beech xylan from pH 8.0 to 10.0 at 68° C: no growth occured in cultures above pH 8.4. Five anaerobic xylanolytic bacteria were isolated from enrichment cultures at pH 8.4; all five microbes were Gram-positive rods with terminal spores, and produced CO₂, H₂, acetate, lactate and ethanol from xylan and xylose. One of the isolates, strain A2, grew from 50 to 75° C, with optimum growth near 68° C, and from pH 5.2 to 9.0 with an optimum between 6.8 and 7.4. Taxonomically, strain A2 was most similar to Clostridium thermohydrosulfuricum. At pH 7.0, the supernatant xylanases of strain A2 had a temperature range from 50 to 78° C with an optimum between 68 and 78° C. At 68° C, xylanase activity occurred from pH 4.9 to 9.1, with an optimum from pH 5.0 to 6.6. At pH 7.0 and 68° C, the K _m of the supernatant xylanases was 2.75 g xylan/l and the V _max was 2.65 × 10^–6 kat/l culture supernatant. When grown on xylose, xylanase production was as high as when grown on xylan. Correspondence to: B. K. Ahring     

Biology of Azospirillum-Sugarcane Association: Enhancement of Nitrogenase Activity

Azospirillum brasilense was reisolated from associations with callus tissue cultures of sugarcane and compared with stock cultures of the inoculated bacterium and related strains. Although the reisolate had a growth rate similar to stock cultures, it exhibited a severalfold increase in maximum specific activity of nitrogenase. The reisolate and the parent culture had similar ultrastructure. The general ultrastructure of Azospirillum is described. The bacterium was capsulated when grown on nitrogen-free nutrient agar plates and on callus, but was not capsulated when growing in a subsurface zone in N-free semisolid nutrient agar, except rarely in aging cultures. Capsulation may be a protective mechanism against unfavorable pO₂ under dinitrogen-fixing conditions. Pleomorphism occurred in capsulated forms, and the ultrastructure of these forms is described.     

Advances in cereal protoplast research

Beginning in 1986, plants have been regenerated from protoplasts of all of the important cereal species, including wheat, rice, maize, and barley, and grasses such as sugarcane. In addition, somatic hybrids/cybrids as well as transgenic plants with introduced useful agronomic traits have been obtained in several instances. This rapid and impressive progress in the genetic manipulation of cereals has been made possible by two critical technical advances during the past decade: the establishment of embryogenic suspension cultures as a source of totipotent protoplasts and the direct delivery of DNA into protoplasts for genetic transformation.     

Hydrogen-bond dynamics of water in presence of an amphiphile,tetramethylurea: signature of confinement-induced effects

Various experimental and simulation studies have suggested that the presence of amphiphilic molecules in aqueous solutions substantially perturbs the tetrahedral hydrogen-bond (H-bond) network of neat liquid water. Such structural perturbation is expected to impact H-bond lifetime of liquid water. Tetramethylurea (TMU) is an example of an amphiphile because it possesses both hydrophobic and hydrophilic moieties. Molecular dynamics simulations of (water+TMU) binary mixtures at various compositions have been performed in order to investigate the microscopic mechanism through which the amphiphiles influence the H-bond dynamics of liquid water at room temperature. Present simulations indicate lengthening of both water–water H-bond lifetime and H-bond structural relaxation time upon addition of TMU in aqueous solution. At the highest TMU mole fraction studied, H-bond lifetime and structural relaxation time are, respectively, ～4 and ～8 times longer than those in neat water. This is comparable with the slowing down of H-bond dynamics for water molecules confined in cyclodextrin cavities. Simulated relaxation profiles are multi-exponential in character at all mixture compositions, and simulated radial distribution functions suggest enhanced water–water and water–TMU interactions upon addition of TMU. No evidence for complete encapsulation of TMU by water H-bond network has been found.     

Ca2+ block and flickering both contribute to the negative slope of the IV curve in BK channels

Single-channel current–voltage (IV) curves of human large-conductance, voltage- and Ca²⁺-activated K⁺ (BK) channels are quite linear in 150 mM KCl. In the presence of Ca²⁺ and/or Mg²⁺, they show a negative slope conductance at high positive potentials. This is generally explained by a Ca²⁺/Mg²⁺ block as by Geng et al. (2013. J. Gen. Physiol. http://dx.doi.org/10.1085/jgp.201210955) in this issue. Here, we basically support this finding but add a refinement: the analysis of the open-channel noise by means of β distributions reveals what would be found if measurements were done with an amplifier of sufficient temporal resolution (10 MHz), namely that the block by 2.5 mM Ca²⁺ and 2.5 mM Mg²⁺ per se would only cause a saturating curve up to +160 mV. Further bending down requires the involvement of a second process related to flickering in the microsecond range. This flickering is hardly affected by the presence or absence of Ca²⁺/Mg²⁺. In contrast to the experiments reported here, previous experiments in BK channels (Schroeder and Hansen. 2007. J. Gen. Physiol. http://dx.doi.org/10.1085/jgp.200709802) showed saturating IV curves already in the absence of Ca²⁺/Mg²⁺. The reason for this discrepancy could not be identified so far. However, the flickering component was very similar in the old and new experiments, regardless of the occurrence of noncanonical IV curves.