Browsing by Subject "Enzymes"
Now showing 1 - 3 of 3
Results Per Page
Sort Options
- ItemOpen AccessThe enzymatic release of glycosidically-bound terpenes in must(1989) Strauss, Lindie Helene; Botes, DPFlavour in wines is perhaps the most important factor affecting wine quality, with monoterpenoids being among the compounds contributing to flavour. In grapes there exists glycosidically-bound forms of these monoterpenes, representing a latent source of aroma. This project is a study on the ability of different enzymes to release these monoterpenes in Muscat d'Alexandrie and Weisser Riesling grapes. Different commercial enzyme preparations were characterised with respect to their catalytic properties and fractionated by ion exchange chromatography in the most active fractions. Amongst those commercial preparations tested, Rohapect C was found to be the most effective at conditions prevalent during wine making. These purified fractions were added to the above mentioned grapes to determine their effect on the release of different monoterpenes. Even though the enzymes were active on synthetic substrates, limited release of terpenes from must could be detected. Possible causes for this apparent inability to release monoterpenes were investigated. Product inhibition due to the presence of high concentrations of glucose in the must appeared to be the main cause of limited enzyme activity. In an attempt to overcome this problem, glucose in the above mentioned cultivars was oxidised to gluconic acid by glucose oxidase, prior to attempting monoterpene release by Rohapect C. Although no marked increase in the release of total terpenes occured, a significant increase in the concentrations of some individual terpenes could be observed. The effect of this on wine quality remains to be ascertained.
- ItemOpen AccessA high-throughput screen against pantothenate synthetase (PanC) identifies 3-biphenyl-4-cyanopyrrole-2-carboxylic acids as a new class of inhibitor with activity against Mycobacterium tuberculosis(Public Library of Science, 2013) Kumar, Anuradha; Casey, Allen; Odingo, Joshua; Kesicki, Edward A; Abrahams, Garth; Vieth, Michal; Masquelin, Thierry; Mizrahi, Valerie; Hipskind, Philip A; Sherman, David RThe enzyme pantothenate synthetase, PanC, is an attractive drug target in Mycobacterium tuberculosis . It is essential for the in vitro growth of M. tuberculosis and for survival of the bacteria in the mouse model of infection. PanC is absent from mammals. We developed an enzyme-based assay to identify inhibitors of PanC, optimized it for high-throughput screening, and tested a large and diverse library of compounds for activity. Two compounds belonging to the same chemical class of 3-biphenyl-4- cyanopyrrole-2-carboxylic acids had activity against the purified recombinant protein, and also inhibited growth of live M. tuberculosis in manner consistent with PanC inhibition. Thus we have identified a new class of PanC inhibitors with whole cell activity that can be further developed.
- ItemOpen AccessRetinoic acid-independent expression of Meis2 during autopod patterning in the developing bat and mouse limb(2015) Mason, Mandy K; Hockman, Dorit; Curry, Lyle; Cunningham, Thomas J; Duester, Gregg; Logan, Malcolm; Jacobs, David S; Illing, NicolaBackgroundThe bat has strikingly divergent forelimbs (long digits supporting wing membranes) and hindlimbs (short, typically free digits) due to the distinct requirements of both aerial and terrestrial locomotion. During embryonic development, the morphology of the bat forelimb deviates dramatically from the mouse and chick, offering an alternative paradigm for identifying genes that play an important role in limb patterning.ResultsUsing transcriptome analysis of developing Natal long-fingered bat (Miniopterus natalensis) fore- and hindlimbs, we demonstrate that the transcription factor Meis2 has a significantly higher expression in bat forelimb autopods compared to hindlimbs. Validation by reverse transcriptase and quantitative polymerase chain reaction (RT-qPCR) and whole mount in situ hybridisation shows that Meis2, conventionally known as a marker of the early proximal limb bud, is upregulated in the bat forelimb autopod from CS16. Meis2 expression is localised to the expanding interdigital webbing and the membranes linking the wing to the hindlimb and tail. In mice, Meis2 is also expressed in the interdigital region prior to tissue regression. This interdigital Meis2 expression is not activated by retinoic acid (RA) signalling as it is present in the retained interdigital tissue of Rdh10trex/trex mice, which lack RA. Additionally, genes encoding RA-synthesising enzymes, Rdh10 and Aldh1a2, and the RA nuclear receptor Rarβ are robustly expressed in bat fore- and hindlimb interdigital tissues indicating that the mechanism that retains interdigital tissue in bats also occurs independently of RA signalling.ConclusionsMammalian interdigital Meis2 expression, and upregulation in the interdigital webbing of bat wings, suggests an important role for Meis2 in autopod development. Interdigital Meis2 expression is RA-independent, and retention of interdigital webbing in bat wings is not due to the suppression of RA-induced cell death. Rather, RA signalling may play a role in the thinning (rather than complete loss) of the interdigital tissue in the bat forelimb, while Meis2 may interact with other factors during both bat and mouse autopod development to maintain a pool of interdigital cells that contribute to digit patterning and growth.Electronic supplementary materialThe online version of this article (doi:10.1186/s13227-015-0001-y) contains supplementary material, which is available to authorized users.