PubMed Health⌕ Search

Biomedical subjects

O Käppeli

Publications and source records attributed to O Käppeli.

30 records · Page 2Linked to original sources

The respirative breakdown of glucose by Saccharomyces cerevisiae: an assessment of a physiological state.

Cells of Saccharomyces cerevisiae exhibiting respirative glucose metabolism in continuous culture were able to use ethanol as a co-substrate. The ethanol uptake rate was dependent on the residual respirative capacity of the cells. The activities of gluconeogenic enzymes and of malate dehydrogenase were higher in cells degrading glucose respiratively than in cells metabolizing glucose respiro-fermentatively, but were lower than in cells growing on ethanol only. The pattern of distribution of the mitochondrial cytochromes was similar but the differences were less distinct. In synchronously growing cells, the activities of gluconeogenic enzymes and of malate dehydrogenase oscillated, with activities increasing during the budding phase. The increase was preceded by the appearance of ethanol in the culture medium.

Cytochromes↗

Structure of the cell surface of the yeast Candida tropicalis and its relation to hydrocarbon transport.

The surface structure of the hydrocarbon-utilizing yeast Candida tropicalis was investigated by scanning and transmission electron microscopy (SEM and TEM respectively). The sample preparation technique was based on a rapid cryofixation without any addition of cryoprotectants. In subsequently freeze-dried samples the surface structure was analysed by scanning electron microscopy. Thin sections were prepared from freeze substituted samples. Both techniques revealed hair-like structures at the surface of hydrocarbon-grown cells. The hairy surface structure of the cells was less expressed in glucose-grown cells and it was absent completely after proteolytic digestion of the cells. When cells were incubated with hexadecane prior to cryofixation a contrast-rich region occurred in the hair fringe of thin sections as revealed by TEM. Since these structures were characteristic for hexadecane-grown cells and could not be detected in glucose-grown or protease-treated cells it was concluded that they originate from hexadecane adhering to the cell surface and are functionally related to hexadecane transport. The structure of the surface and its relation to hydrocarbon transport are discussed in view of earlier results on the chemical composition of the surface layer of the cell wall.

Candida↗

The cytochrome P450-containing monooxygenase of Trichosporon cutaneum: occurrence and properties.

Trichosporon cutaneum metabolizes glucose purely oxidatively and cytochrome P450 was not detected in the reduced CO-difference spectrum of whole cells. However, in the isolated microsomal fraction the corresponding monooxygenase was present as shown by the appearance of cytochrome P450, NADPH-cytochrome c(P450) reductase and cytochrome b5. The absorption maximum of the terminal oxidase in the reduced CO-difference spectrum shifted between 447 and 448 nm. Derepression of biosynthesis of all components was achieved by transition of the cells from carbon- to oxygen-limited growth in continuous culture. The monooxygenase exhibited aminopyrine demethylation activity but not omega-hydroxylation activity of lauric acid. With respect to the growth limiting nutrient (carbon and oxygen respectively), mitochondrial cytochrome content showed an analogous behavior as cytochrome P450 and cytochrome b5.

Candida↗

Pseudomonas aeruginosa biosurfactant production in continuous culture with glucose as carbon source.

Rsan-ver, a strain of Pseudomonas aeruginosa isolated at this department, was used for the development of a continuous process for biosurfactant production. The active compounds were identified as rhamnolipids. A final medium for production was designed in continuous culture by means of medium shifts, since the formation of surface-active compounds was decisively influenced by the composition and concentration of the medium components. In the presence of yeast extract, biosurfactant production was poor. For the nitrogen-source nitrate, which was superior to ammonium, an optimum carbon-to-nitrogen ratio of ca. 18 existed. The iron concentration needed to be minimized to 27.5 micrograms of FeSO4 X 7H2O per g of glucose. A carbon-to-phosphate ratio below 16 yielded the maximum production of rhamnolipids. The final productivity dilution rate diagram indicated that biosurfactant production was correlated to low growth rates (dilution rate below 0.15 h-1). With a medium containing 18.2 g of glucose liter-1, a biosurfactant concentration (expressed as rhamnolipids) of up to 1.5 g liter-1 was obtained in the cell-free culture liquid.

Ammonium Sulfate↗

Metabolic conditions determining the composition and catalytic activity of cytochrome P-450 monooxygenases in Candida tropicalis.

In the microsomal fraction of Candida tropicalis cells, two distinct monooxygenases were detected, depending on the growth conditions. The distinction of the two monooxygenases was evident from: (i) the absorption maxima in the reduced CO difference spectra of the terminal oxidases (cytochromes P-450 and P-448); (ii) the contents of the monooxygenase components (cytochromes P-450/P-448, NADPH-cytochrome c (P-450) reductase, and cytochrome b5) and (iii) the catalytic activity of the complete system (aliphatic hydroxylation and N-demethylation activity). The occurrence of the respective monooxygenases could be related to the carbon source (n-alkanes or glucose). Oxygen limitation led to a significant increase of cytochrome P-450/P-448 content, independent of the carbon source utilized by the cells. An improved method for the isolation of microsomes enabled us to demonstrate the presence of cytochrome P-448 in glucose-grown cells.

Alkanes↗

Partition of alkane by an extracellular vesicle derived from hexadecane-grown Acinetobacter.

The enhanced solubility of hexadecane in the growth medium of hexadecane-grown Acinetobacter species has been related to the accumulation of an extracellular vesicular component. The partition of hexadecane was determined by measuring the amount of [3H]hexadecane bound to the vesicular particle. The vesicle was characterized as a phospholipid-rich, lipopolysaccharide-rich particle with a polypeptide composition similar to the outer membrane of Acinetobacter. The accumulation of an extracellular vesicular component that binds hexadecane in the form of a microemulsion represents another example of molecules produced by microorganisms in response to paraffinic substrates.

Acinetobacter↗

Chemical and structural alterations at the cell surface of Candida tropicalis, induced by hydrocarbon substrate.

The surface-localized polysaccharide of alkane-grown cells of Candida tropicalis was identified as mannan containing approximately 4% covalently linked fatty acids. Glucose-grown cells lacked the mannan-fatty acid complex. The surface structure of alkane-grown cells showed a radial arrangement of the wall polymers, with protruding parts. The cell surface of glucose-grown cells was smooth, with a coherent outer limit. The mannan was localized by using concanavalin A. Masking of the mannan with concanavalin A reduced the binding affinity of the surface for alkane, indicating the involvement of the surface-localized mannan-fatty acid complex in the binding of alkanes.

Alkanes↗

Component from the cell surface of the hydrocarbon-utilizing yeast Candida tropicalis with possible relation to hydrocarbon transport.

A polysaccharide-fatty acid complex was isolated from the cell surface of Candida tropicalis growing on alkanes. This complex was solubilized by Pronase treatment of whole cells. A decrease in alkane-binding affinity was observed after Pronase treatment, resulting in 10 to 12% of the yeast dry cell weight being released as polysaccharide. The isolated polysaccharide contained 2.5% fatty acids. C. tropicalis and Saccharomyces cerevisiae grown with glucose contained only traces of fatty acids in the corresponding polysaccharide fraction. The fatty acids were not removed from the polysaccharide moiety by gel filtration. Extraction of the polysaccharide with chloroform-methanol showed that fatty acids were covalently bound to the polysaccharide. The amphipathic nature of the isolated polysaccharide and the hydrocarbon-induced formation suggest a possible role in alkane metabolism.

Alkanes↗