Enzymatic hydrolysis of inulin to fructose by glutaraldehyde fixed yeast cells

Inulin, a polyfruction, is found as the reserve carbohydrate in the roots and tubers of various plants (i.e. Jerusalem artichoke, chicory, and dahlia tubers). The beta-fructofuranosidase (inulase) from the yeast Kluyveromyces fragilis is of interest because of its industrial potential in fructose syrup and alcohol production from inulin containing plants. We have found that the inulase of K. fragilis can be immobilized in the yeast cells by glutaraldehyde treatment. These cells are resistant to physical and enzymatic destruction. Although the exact nature of the immobilization is not fully understood, the kinetic parameters of the immobilized enzyme are similar to those of the soluble enzyme. No reduction of enzyme activity was observed after glutaraldehyde treatment and glutaraldehyde concentration did not affect enzyme activity. A 96% hydrolysis of dahlia inulin was achieved in 10.5 h with a 9.5% (w/v) fixed enzyme suspension. A Jerusalem artichoke extract containing 16.8%polyfructan was completely hydrolyzed in 3.5 h with a 0.24% (w/v)fixed enzyme suspension. This is a time frame feasible for industrial consideration.     

Identification of a nuclear localization signal and importin beta members mediating NUAK1 nuclear import inhibited by oxidative stress

NUAK1 is a serine/threonine kinase member of the AMPK-α family. NUAK1 regulates several processes in tumorigenesis; however, its regulation and molecular targets are still poorly understood. Bioinformatics analysis predicted that the majority of NUAK1 localizes in the nucleus. However, there are no studies about the regulation of NUAK1 subcellular distribution. Here, we analyzed NUAK1 localization in several human cell lines, mouse embryo fibroblasts, and normal mouse tissues. We found that NUAK1 is located in the nucleus and also in the cytoplasm. Through bioinformatics analysis and studies comparing subcellular localization of wild type and NUAK1 mutants, we identified a conserved bipartite nuclear localization signal at the N-terminal domain of NUAK1. Based on mass spectrometry analysis, we found that NUAK1 interacts with importin-β members including importin-β1 (KPNB1), importin-7 (IPO7), and importin-9 (IPO9). We confirmed that importin-β members are responsible for NUAK1 nuclear import through the inhibition of importin-β by Importazole and the knockdown of either IPO7 or IPO9. In addition, we found that oxidative stress induces NUAK1 cytoplasmic accumulation, indicating that oxidative stress affects NUAK1 nuclear transport. Thus, our study is the first evidence of an active nuclear transport mechanism regulating NUAK1 subcellular localization. These data will lead to investigations of the molecular targets of NUAK1 according to its subcellular distribution, which could be new biomarkers or targets for cancer therapies.     

‘Pollen potency’: the relationship between atmospheric pollen counts and allergen exposure

Pollen allergies are responsible for a considerable global public health burden, and understanding exposure is critical to addressing the health impacts. Atmospheric pollen counts are routinely used as a predictor of risk; however, immune responses are triggered by specific proteins known as allergens, which occur both within and on the surface of the pollen grain. The ratio between atmospheric pollen counts and allergen concentrations (‘pollen potency’) has been shown to be inconsistent, with potentially important implications for pollen monitoring practice. Despite this, there has been no previous synthesis of the literature and our understanding of the factors that influence pollen potency remains poor. We conducted a scoping review with the aim of deriving a current understanding of: (a) the factors that influence pollen potency; (b) its variation through time, between taxa and by location; and (c) the implications for pollen monitoring practice. Our synthesis found that pollen potency is highly variable within and between seasons, and between locations; however, much of this variability remains unexplained and has not been deeply investigated. We found no predictable pollen potency patterns relating to taxon, geography or time, and inconclusive evidence regarding possible driving factors. With respect to human health, the studies in our synthesis generally reported larger associations between atmospheric allergen loads and allergy symptoms than whole pollen counts. This suggests that pollen potency influences public health risk; however, the evidence base remains limited. Further research is needed to better understand both pollen potency variability and its implications for health.

     

Alkaloids of Argemone fruticosa and A. echinata

Argemone fruticosa Thurber ex Gray was analyzed and found to contain hunnemanine as the major alkaloid along with allocryptopine. A. echinata G. B. Ownb. was found to contain cryptopine and berberine as major alkaloids. These analyses indicate that these two species belong among the more specialized species of the genus.