Universität Stuttgart
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Item Open Access Die Spaltung von Arylether-Bindungen durch initiale Dioxygenierung: Grundlage des bakteriellen Dioxinabbaus(1991) Engesser, Karl-Heinrich; Strubel, Volker; Kirchner, S.; Schestag, S.; Schulte, P.; Knackmuss, Hans-JoachimBei der Untersuchung des bakteriellen Abbaus von Arylether-Modellsubstraten wie 2-Alkoxybenzoat, Carboxybiphenylether und Dibenzofuran wurde ein grundlegender Mechanismus für die Spaltung von Aryletherbindungen aufgedeckt. Demnach bewirken Dioxygenase-Enzyme unter Einführung zweier Hydroxylgruppen die Überführung von Ether- in Hemiacetalbindungen. Diese instabilen Hemiacetale reagieren unter Rearomatisierung zu aliphatischen Alkoholen und/oder Phenolverbindungen ab. Enzyme dieses Typs sind auch in der Lage, Dioxine zu spalten.Item Open Access Microbial metabolism of chlorosalicylates: effect of prolonged subcultivation on constructed strains(1986) Rubio, Miguel Angel; Engesser, Karl-Heinrich; Knackmuss, Hans-JoachimThe hybrid strain Pseudomonas sp. WR4016 was subcultivated with increasing concentrations of 5-chlorosalicylate (5rarr10 mM) as sole carbon source over a period of 9 months. At intervals of approximately 3 months derivative strains WR4017, WR4018 and WR4019 were isolated which exhibited higher growth rates and increased substrate tolerance. Comparative analysis of the turnover rates of the key enzymes in chlorosalicylate degradation showed that the adaptation process did not result from structural modifications of these proteins. Instead, balanced over-production of the salicylate hydroxylase and catechol 1,2-dioxygenase prevented the accumulation of toxic chlorocatechols and accounted for the reduction of the doubling times with 4- or 5-chlorosalicylate. A comparative analysis of a genetically engineered chlorosalicylate degrader PL300-1 showed similar regulatory patterns as the most advanced isolate WR4019 from the adaptation series.Item Open Access Metabolism of 2-chloro-4-methylphenoxyacetate by Alcaligenes eutrophus JMP 134(1993) Pieper, Dietmar Helmut; Stadler-Fritzsche, Karin; Engesser, Karl-Heinrich; Knackmuss, Hans-Joachim2-Chloro-4-methylphenoxyacetate is not a growth substrate for Alcaligenes eutrophus JMP 134 and JMP 1341. It is, however, being transformed by enzymes of 2,4-dichlorophenoxyacetic acid metabolism to 2-chloro-4-methyl-cis, cis-muconate, which is converted by enzymatic 1,4-cycloisomerization to 4-carboxymethyl-2-chloro-4-methylmuconolactone as a dead end metabolite. Chemically, only 3,6-cycloisomerization occurs, giving rise to both diastereomers of 4-carboxychloromethyl-3-methylbut-2-en-4-olide. Those lactones harbonring a chlorosubstituent on the 4-carboxymethyl side chain were surprisingly stable under physiological as well as acidic conditions.Item Open Access Degradation of 2-bromo-, 2-chloro- and 2-fluorobenzoate by Pseudomonas putida CLB 250(1989) Engesser, Karl-Heinrich; Schulte, P.Pseudomonas putida strain CLB 250 (DSM 5232) utilized 2-bromo-, 2-chloro- and 2-fluorobenzoate as sole source of carbon and energy. Degradation is suggested to be initiated by a dioxygenase liberating halide in the first catabolic step. After decarboxylation and rearomatization catechol is produced as a central metabolite which is degraded via the ortho-pathway. After inhibition of ring cleavage activities with 3-chlorocatechol, 2-chlorobenzoate was transformed to catechol in nearly stoichiometric amounts. Other ortho-substituted benzoates like anthranilate and 2-methoxybenzoate seem to be metabolized via the same route.Item Open Access Kombination aus Biowäscher- und Biomembranverfahren zur Reinigung von Abluft und hydrophilen und hydrophoben Inhaltsstoffen(1994) Reiser, Martin; Fischer, Klaus; Engesser, Karl-HeinrichAbluft, die eine Mischung aus wasserlöslichen uod wasserunlöslichen organischen Lösungsmitteln enthält, stellt für ein biologisches Reinigungsverfahren eine große Herausforderung dar. Durch Kombination eines Biomembranreaktors mit Silikonkautschukmembranen und einem Biowäscher konnten für ein Gemisch von Methanol, n-Hexan und Toluol in der Abluft gute Reinigungsleistungen erzielt werden.Item Open Access Einsatz von Gensonden zur Identifikation von Mikroorganismen im belebten Schlamm(1994) Engesser, Karl-Heinrich; Schmid, AndreasDie Gensondentechnik stellt heute schon einen extrem leistungsfähigen, neuartigen Ansatz zur Populationsanalyse komplexer Ökosysteme und damit auch der Charakterisierung der "Biomasse" des Belebtschlammes dar. Für die Gensondentechnik typisch ist ein relativ geringer Zeitaufwand bei der Durchführung. Dies ermöglicht erstmals die Analyse der Dynamik einer Population nahezu in Echtzeit.Item Open Access Assemblage of ortho cleavage route for simultaneous degradation of chloro- and methylaromatics(1987) Rojo, Fernando; Pieper, Dietmar H.; Engesser, Karl-Heinrich; Knackmuss, Hans-Joachim; Timmis, Kenneth N.Genetic engineering is a powerful means of accelerating the evolution of new biological activities and has considerable potential for constructing microorganisms that can degrade environmental pollutants. Critical enzymes from five different catabolic pathways of three distinct soil bacteria have been combined in patchwork fashion into a functional ortho cleavage route for the degradation of methylphenols and methylbenzoates. The new bacterium thereby evolved was able to degrade and grow on mixtures of chloro- and methylaromatics that were toxic even for the bacteria that could degrade the individual components of the mixtures. Except for one enzymatic step, the pathway was fully regulated and its component enzymes were only synthesized in response to the presence of pathway substrates.Item Open Access Emission von Pilzsporen in Müllverarbeitungsanlagen(1994) Göttlich, Elke; Engesser, Karl-Heinrich; Bardtke, DieterDie Häufigkeit und Sporendichte verschiedener Pilzarten in der Luft von Kompostwerken, bei der Sortierung, Müllverbrennung und auf Deponien wurden in einer Studie in 11 Betrieben untersucht. Außer A. fumigatus, über dessen klinische und allergologische Bedeutung die meisten Kenntnisse vorliegen, wurden verschiedene Pilzarten, die als potentiell gesundheitlich relevant angesehen werden,in Keimdichten über den derzeitigen Orientierungswerten für Pilzbelastungen der Luft nachgewiesen. Obwohl vergleichbare Daten zumeist fehlen und daher gesicherte Richtgrenzwerte gegenwärtig nicht festgelegt werden können, sind die gesundheitlichen Risiken der Beschäftigten an den Arbeitsplätzen als erheblich anzusehen.Item Open Access Enrichment of dibenzofuran utilizing bacteria with high co-metabolic potential towards dibenzodioxin and other anellated aromatics(1989) Strubel, Volker; Rast, Hans G.; Fietz, Walter H.; Knackmuss, Hans-Joachim; Engesser, Karl-HeinrichDibenzofuran degrading bacteria were enriched from various environmental sources. A mutualistic mixed culture of strain DPO 220 and strain DPO 230 was characterized. Strain DPO 220 alone showed limited growth with dibenzofuran as sole source of carbon and energy (td ≥ 4.5 h). A labile degradation product, C12H10O5, and salicylate were isolated from the culture fluid. Salicylate was found to be a central intermediate of DBF-degradation.Strain DPO 220 co-metabolized a wide range of anellated aromatics as well as heteroaromatics. High rates of co-oxidation of dibenzodioxin demonstrate analogue-enrichment to be a powerful technique for selecting enzymatic activities for otherwise non-degradable substrates.Item Open Access Simultaneous degradation of chloro- and methylaromatics via ortho pathway by genetically engineered bacteria and natural soil isolates(1989) Engesser, Karl-Heinrich; Pieper, Dietmar H.; Rojo, Fernando; Timmis, Kenneth N.; Knackmuss, Hans-JoachimThe simultaneous bacterial metabolism of chloro- and methylaromatics via ortho- or metapathway, normally results in incomplete degradation and death of the organisms. This is caused by misrouting of central intermediates. i.e. substituted catechols into unproductive pathways and suicide inactivation of the key enzyme of meta pathway, (catechol 2,3-dioxygenase). The meta pathway proved to be definitely unsuited for productive metabolism of chloroaromatics. Therefore two strategies were used for simultaneous degradation of mixtures of chloro- and methylaromatics via ortho pathways: Methyllactons or certain mixtures of chloro- and methylaromatics were used as enrichment substrates, yielding strains which metabolized these compounds almost exclusively via the desired pathway. Alternatively relevant enzymes from five different catabolic pathways of three distinct soil bacteria were combined in a patchwork fashion generating a functional ortho cleavage route for methylaromatics coexisting with the ortho cleavage pathway of chloroaromatics.