Regulation of enzymes and isoenzymes of carbohydrate metabolism in the yeast Saccharomyces cerevisiae

An electrophoretic method has been devised to investigate the changes in the enzymes and isoenzymes of carbohydrate metabolism, upon adding glucose to derepressed yeast cell. (i) Of the glycolytic enzymes tested, enolase II, pyruvate kinase and pyruvate decarboxylase were markedly increased. This increase was accompanied by an overall increase in glycolytic activity and was prevented by cycloheximide, an inhibitor of protein synthesis. (ii) In contrast, respiratory activity decreased after adding glucose. This decrease was clearly shown to be the result of repression of respiratory enzymes. A rapid decrease within a few minutes of adding glucose, by analogy with the so-called ‘Crabtree effect’, was not observed in yeast. (iii) The gluconeogenic enzymes, fructose-1,6-bisphosphatase and malate dehydrogenase, which are inactivated after adding glucose, showed no significant changes in electrophoretic mobilities. Hence, there was no evidence of enzyme modifications, which were postulated as initiating degradation. However, it was possible to investigate cytoplasmic and mitochondrial malate dehydrogenase isoenzymes separately. Synthesis of the mitochondrial isoenzyme was repressed, whereas only cytoplasmic malate hydrogenase was subject to glucose inactivation.     

Untersuchungen über den osmotischen Wert polyploider Pflanzen

Zusammenfassung In der Mehrzahl der untersuchten Fälle (insgesamt über 1000 Einzelmessungen) wurden die osmotischen Werte der Tetraploiden niedriger gefunden als die der Diploiden, doch sind die Unterschiede im allgemeinen gering. Junge Pflanzen hatten immer niedrigere 4n-Werte; bei alten Pflanzen und bei Pflanzen unter extremen Kulturbedingungen waren aber vielfach die Werte der 4n höher. Es wird die Hypothese aufgestellt, daß niedrigere osmotische Werte der Tetraploiden den Normalzustand darstellen, daß aber die Polyploiden wie in anderen Eigenschaften, so auch in ihrer Osmotik variabler sind und unter dem Einfluß von Außen-bedingungen ihre osmotischen Werte stärker modifizieren.Unter physiologischen Zwangsbedingungen (Trockenkultur, Pfropfungen) wurde an den Tetraploiden eine unter den gegebenen Verhältnissen zweckmäßige Erhöhung ihrer osmotischen Werte über das Niveau der 2n beobachtet. Gegen Trockenklima erwiesen sich die untersuchten Tetraploiden im Vergleich mit den Diploiden resistenter.Irgendwelche artspezifischen Eigentümlichkeiten konnten im Rahmen der untersuchten Pflanzen nicht festgestellt werden.Mit 6 Textabbildungen.     

Anwendung der automatisierten Festphasenextraktion bei der Bestimmung von Ochratoxin A

For some foodstuffs, determination of the mycotoxin ochratoxin A (OTA) requires time consuming clean up by means of solid phase extraction (SPE). Therefore a system for automated SPE was tested for cleaning up roasted coffee as a possible way of shortening preparation time. Validation of the method in accordance to the so called “Concept '98” led to a LOD of 0.2 μg/kg and a recovery rate of 92%. By using the described procedure with samples of roasted coffee the OTA contents varied between the LOD and 3.4 μg/kg. This method was also used to determine ochratoxin A in liquorice roots, ginger and valerian.

Presented at the 26^th Mykotoxin Workshop in Herrsching, Germany, May 17–19, 2004     

Bacteriophage-resistance mechanisms examined by transfection of Lactococcus lactis subsp. lactis 4513-5 mediated by high voltage electroporation

Summary Phage adsorption tests and transfection by electroporation were carried out to decide whether phage-resistance in Lactococcus lactis subsp. lactis strain 4513-5 is based on intracellular or extracellular mechanisms. Using high voltage (12.5 kV/cm) electroporation, untreated phage DNA was introduced into phage-sensitive and phage-resistant cells. Since phages showed low adsorption frequencies on resistant bacteria, resistance is localized in the cell wall preventing phage DNA from entering the cell. This is the only mechanism responsible for the resistance of L. lactis subsp. lactis 4513-5 against its homologous phage P4513-K12 and non-homologous phages P05M-13 and P05M-47, but not against phage P530-7 and phage P530-12. In the case of the latter two phage strains, intracellular resistance mechanisms are involved and discussed.     

Continuous coenzyme dependent stereoselective synthesis of sulcatol by alcohol dehydrogenase

Summary 6-methyl-5-hepten-2-one was reduced to sulcatol ((+)-6-methyl-5-hepten-2-ol) by using alcohol dehydrogenase fromThermoanaerobium brockii in a continuous process. The cofactor NADP(H) was retained by a charged UF-membrane and regenerated by oxidation of isopropanol to acetone. Use of native NADP in a charged UF-membrane reactor proved to be superior to use of PEG coupled NADP in a uncharged UF-membrane reactor.     

Immunohistological detection of Saruplase (recombinant single-chain urokinase-type plasminogen activator) in normal rat tissue

Summary Saruplase — a recombinant single-chain urokinase-type plasminogen activator was identified immunohistochemically in normal rat tissue after intravenous administration by means of a polyclonal antibody. For this purpose, rat tissues were fixed in various ways (liquid nitrogen, ethanol, formaldehyd solution). Saruplase could be detected by the PAP method, streptavidinbiotin system and indirect immunofluorescence in the kidney (proximal tubule), liver (hepatocytes, Kupffer cells) and spleen (reticular cells). Saruplase was not localized in the rat endothelium. It is discussed that the ratspecific receptors for urokinase-type plasminogen activator on endothelial cells cannot bind Saruplase due to the extreme species specificity.