Acetylsalicylic acid as antifungal in <Emphasis Type="Italic">Eremothecium</Emphasis> and other yeasts

Interesting distribution patterns of acetylsalicylic acid (ASA, aspirin) sensitive 3-hydroxy (OH) oxylipins were previously reported in some representatives of the yeast genus Eremothecium—an important group of plant pathogens. Using immunofluorescence microscopy and 3-OH oxylipin specific antibodies in this study, we were able to map the presence of these compounds also in other Eremothecium species. In Eremothecium cymbalariae, these oxylipins were found to cover mostly the spiky tips of narrowly triangular ascospores while in Eremothecium gossypii, oxylipins covered the whole spindle-shaped ascospore with terminal appendages. The presence of these oxylipins was confirmed by chemical analysis. When ASA, a 3-OH oxylipin inhibitor, was added to these yeasts in increasing concentrations, the sexual stage was found to be the most sensitive. Our results suggest that 3-OH oxylipins, produced by mitochondria through incomplete β-oxidation, are associated with the development of the sexual stages in both yeasts. Strikingly, preliminary studies on yeast growth suggest that yeasts, characterized by mainly an aerobic respiration rather than a fermentative pathway, are more sensitive to ASA than yeasts characterized by both pathways. These data further support the role of mitochondria in sexual as well as asexual reproduction of yeasts and its role to serve as a target for ASA antifungal action.     

Epoxide hydrolase activity of Chryseomonas luteola for the asymmetric hydrolysis of aliphatic mono-substituted epoxides

Asymmetric hydrolysis of a homologous range of straight chain 1,2-epoxyalkanes was achieved using whole cells of Chryseomonas luteola. Depending on the chain length, hydrolyses of the racemic epoxides afforded optically active epoxides and diols with varying degrees of optical purity. In the case of 1,2-epoxyoctane, the enantiomeric excess of the remaining (S)-epoxide and formed (R)-diol was excellent (ees > 98% and eep = 86%). This is the first report of a bacterial epoxide hydrolase with such unusual enantioselectivity for terminal mono-substituted epoxides bearing no directing group on the chiral C-2 carbon. Benzyl glycidyl ether and the 2,2-disubstituted epoxide, 2-methyl-1,2-epoxyheptane, were hydrolysed, but no enantioselectivity was observed. © Rapid Science Ltd. 1998     

Encapsulation of Lactobacillus plantarum 423 and its Bacteriocin in Nanofibers

Plantaricin 423, produced by Lactobacillus plantarum 423, was encapsulated in nanofibers that were produced by the electrospinning of 18% (w/v) polyethylene oxide (200 000 Da). The average diameter of the nanofibers was 288 nm. Plantaricin 423 activity decreased from 51 200 AU/ml to 25 600 AU/ml and from 204 800 AU/ml to 51 200 AU/ml after electrospinning, as determined against Lactobacillus sakei DSM 20017 and Enterococcus faecium HKLHS, respectively. Cells of L. plantarum 423 encapsulated in nanofibers decreased from 2.3 × 10¹⁰ cfu/ml before electrospinning to 4.7 × 10⁸ cfu/ml thereafter. Cells entrapped in the nanofibers continued to produce plantaricin 423. This is the first report on the encapsulation of a bacteriocin and cells of L. plantarum in nanofibers. The method may be used to design a drug delivery system for bacteriocins and the encapsulation of probiotic lactic acid bacteria. The technology is currently being optimized.     

The production of γ-linolenic acid by selected members of the Dikaryomycota grown on different carbon sources

In this study, seven fungal strains, representing different phylogenetic groups within the Dikaryomycota, were tested for the presence of -linolenic acid [18:3(6)], when grown in synthetic liquid media devoid of fatty acids, on a series of 40 different carbon sources. The fungal strains represented the species Dipodascopsis uninucleata, Eurotium rubrum, Galactomyces geotrichum, Neurospora crassa, Saccharomyces cerevisiae, Spongipellis unicolor and Talaromyces flavus. Cultures were periodically harvested during growth and the fatty acids in the total lipids analysed as methyl esters, using gas chromatography and mass spectrometry. It was found that 18:3(6) is present in E. rubrum CBS 350.65, S. unicolor CBS 117.16 and in T. flavus CBS 310.38NT, when these strains were grown on certain carbon sources. No correlation between the growth phase of the organism and the presence of 18:3(6) could be detected. In order to confirm the production of 18:3(6), the lipid metabolism of two unrelated dikaryomycotan fungi (S. unicolor CBS 117.16 and E. rubrum CBS 350.65) grown on two different carbon sources each, was examined. Cultures of E. rubrum CBS 350.65 were grown on glucose and sorbose and cultures of S. unicolor CBS 117.16 on glucose and sucrose in synthetic liquid media with a C:N ratio of 50:1 (w/w). The total lipids of these cultures were fractionated and the fatty acids in the fractions analysed as methyl esters, using gas chromatography and mass spectrometry. The lipid metabolism of both E. rubrum CBS 350.65 and S. unicolor CBS 117.16 differed on the two carbon sources used. The ab initio production of 18:3(6) by E. rubrum CBS 350.65 in synthetic liquid media was confirmed. In contrast, the ab initio production of 18:3(6) by S. unicolor CBS 117.16 in synthetic liquid media could not be confirmed.     

Ants, altitude and change in the northern Cape Floristic Region

Aim Climate‐modelling exercises have demonstrated that the Cape Floristic Region is highly sensitive to climate change and will apparently lose much of its northern limits over the next few decades. Because there is little monitoring of diversity in this area, ant assemblage structure was investigated within the main vegetation types in the Greater Cederberg Biodiversity Corridor. In particular, we sought to determine how ant assemblage structure differs between the main vegetation types, how restricted ants – and in particular the major myrmecochores – are to the major vegetation types, and which environmental variables might underlie differences in the ant assemblages and in the specificity of species to particular areas. Location Northern Cape Floristic Region, Western Cape, South Africa. Methods Sampling was undertaken during October 2002 and March 2003 across an altitudinal gradient ranging from sea level (Lambert's Bay) to c. 2000 m a.s.l. (Sneeukop, Cederberg) and down again to 500 m a.s.l. (Wupperthal) in the Western Cape, South Africa. Pitfall traps were used to sample ants at 17 altitudinal bands, stretching over three vegetation types (Strandveld, Mountain Fynbos and Succulent Karoo). Biotic and abiotic environmental variables were collected at each sampling site. Generalized linear models were used to determine the relationships between species richness, density, abundance and the abundance of the major myrmecochores, and the environmental variables. Redundancy analysis was used to determine the relationship between ant assemblage structure and the environmental variables. The Indicator Value Method was used to identify characteristic ant species for each vegetation type and altitudinal site. Results Temperature explained significant proportions of the variation in species density and abundance, and, together with area and several vegetation variables, contributed significantly to the separation of the assemblages in the major vegetation types and biomes. Four major myrmecochores were identified [Anoplolepis sp. (cf. custodiens), Anoplolepis sp. (cf. steinergroeveri), Camponotus niveosetosus, Tetramorium quadrispinosum]. The abundances of the two Anoplolepis species were related to vegetation variables, while the abundance of the other two species showed opposite relationships with temperature variables. Fourteen ant species were characteristic of certain vegetation types and altitudes. Several of these species contributed to the differences between the assemblages. Main conclusions There are likely to be substantial and complex changes to ant assemblages as climates change in the northern Cape Floristic Region. Moreover, the importance of ants for ecosystem functioning suggests that these responses are not only likely to be a response solely to vegetation changes, but might also precipitate vegetation changes. The changes that are predicted to take place in the next 50 years in the Cape Floristic Region could be substantially exacerbated by such synergistic effects, which have major implications for long‐term conservation plans. Ongoing monitoring of this transect will reveal the nature and pace of the change as it unfolds.     

Cloning and sequencing of an epoxide hydrolase gene from Rhodosporidium paludigenum.

Epoxide hydrolase (EH) activity was recently described in yeasts and highly selective hydrolysis of epoxides was observed during whole cell biotransformations. To expand the available molecular data regarding yeast EHs, the EH encoding gene from Rhodosporidium paludigenum (CBS 6565) was isolated, cloned and sequenced. The genomic EH sequence revealed a 1600 bp sequence interrupted by six introns. cDNA sequence analysis revealed an open reading frame of 1236 bp with a deduced polypeptide length of 411 amino acids. The deduced amino acid sequence revealed a relative high degree of sequence homology compared to the amino acid sequence of the EH from Rhodotorula glutinis.     

The long-chain fatty acid compositions of species representing the genus Kluyveromyces

Abstract The cellular long-chain fatty acids of 32 strains representing 10 species of the genus Kluyveromyces were extracted by saponification and analyzed as methyl esters by gas chromatography. The Kluyveromyces strains were characterized by the presence of palmitate, palmitoleate, oleate, and linoleic acid as the major fatty acids. These strains were divided into 3 groups on the basis of fatty acid content. The first group was characterized by a high percentage of linolenic acid, the second group of a lower percentage and the third group by the absence of linolenic acid.     

A molecular approach to the characterization of an industrialBacillus amyloliquefaciens strain

Classification of an industrial strain ofBacillus from which we have previously cloned an -amylase gene yielded ambiguous results with classical methods. However, positive identification asB. amyloliquefaciens was obtained by DNA analysis, by use of restriction enzyme fingerprinting and DNA hybridization analysis of genomic DNA, and by characterization of the purified -amylase.