Showing papers in "Fems Microbiology Letters in 1997"

Bacillus cereus and its food poisoning toxins

Abstract: Bacillus cereus is becoming one of the more important causes of food poisoning in the industrialised world. It produces one emetic toxin and three different enterotoxins. The emetic toxin is a ring-shaped structure of three repeats of four amino and/or oxy acids: [d-O-Leu-d-Ala-l-O-Val-l-Val]3. This ring structure has a molecular mass of 1.2 kDa, and is chemically closely related to the potassium ionophore valinomycin. Two of the three enterotoxins have been shown to be involved in food poisoning. They both consist of three different proteins that act together. One of these enterotoxins is also a haemolysin. This haemolytic enterotoxin is transcribed from one operon. The third enterotoxin is a single component protein, but has not been shown to be involved in food poisoning.

Acid stress responses in enterobacteria

Abstract: The enteric microorganisms Salmonella typhimurium, Escherichia coli and Shigella flexneri prefer to grow in neutral pH environments. They nevertheless experience dramatic pH fluctuations in nature and during pathogenesis. In response to environmental encounters with acid, these organisms have evolved complex, inducible acid survival strategies. Regulatory features include an alternative sigma factor (σS), 2-component signal transduction systems (PhoP/Q; MviA/?) and the major iron regulatory protein Fur. Specific survival mechanisms include emergency pH homeostasis by inducible amino acid decarboxylases and probable roles for DNA repair, chaparonins, membrane biogenesis as well as others that remain poorly defined. Continued study of acid survival in these organisms will provide general insights regarding stress management and will have a direct impact on our understanding of pathogenesis.

High efficiency intergeneric conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA-restricting streptomycetes

Abstract: Many streptomycetes, including S. coelicolor A3(2), possess a potent methyl-specific restriction system which can present an effective barrier to the introduction of heterologous DNA. We have compared the efficiency of intergeneric conjugal transfer of different types of plasmids to S. coelicolor and S. lividans 66 using two E. coli donors: the standard, methylation proficient strain S17-1. and the methylation deficient donor, ET12567(pUB307). We demonstrate that the methylation deficient donor can yield > 104-fold more S. coelicolor exconjugants than the standard donor. In the case of pSET152 derivatives, which integrate into the host chromosome by site-specific recombination, up to 10% of streptomycete spores in the conjugation mixture inherit the plasmid. The conjugation procedure is efficient enough to obtain exconjugants with ‘suicide’ delivery plasmids and therefore provides a simple route for conducting gene disruptions in methyl DNA-restricting streptomycetes, and possibly other bacteria.

PCR-RFLP analysis of the Cryptosporidium oocyst wall protein (COWP) gene discriminates between C. wrairi and C. parvum, and between C. parvum isolates of human and animal origin

Abstract: Cryptosporidium wrairi was isolated from guinea pigs during a spontaneous outbreak of cryptosporidiosis. Despite the morphological and antigenic similarities to C. parvum, C. wrairi displayed a different host range and site of infection and may represent a separate species or sub-species. We used the polymerase chain reaction to clone two distinct 550 bp-long DNA fragments, Wc-I and Wc-II, of the gene encoding the Cryptosporidium oocyst wall protein (COWP) of C. wrairi, which showed 98% identity to the C. parvum homologue. Within Wc-I, polymorphic RsaI restriction sites were used to develop a polymerase chain reaction-restriction fragment length polymorphism method able to distinguish C. wrairi from C. parvum and to identify two groups of C. parvum isolates differentially associated with animal and human infections.

Gene replacement in Staphylococcus carnosus and Staphylococcus xylosus

Abstract: A system for high-efficiency gene replacement in Staphylococcus carnosus and Staphylococcus xylosus has been developed, that is based on temperature-sensitive Escherichia coli-Staphylococcus shuttle vectors for fragment delivery and erythromycin resistance cassettes to facilitate selection of genomic copies of disrupted genes. The approach was tested by constructing a phosphotransferase-deficient mutant of S. carnosus and an S. xylosus mutant strain unable to utilize sucrose. Allelic replacements were observed at rather high frequencies, ranging from approximately 10% for the ptsI gene in S. carnosus up to 50% for the scrB gene in S. xylosus. These differences most likely reflect the length of homology rather than strain-specific variations in recombination efficiencies. Apart from the staphylococcal species tested in this study, the system appears to be applicable in other staphylococci.

Evidence of autoinducer activity in naturally occurring biofilms

Abstract: N-Acyl homoserine lactone (AHL) molecules have been shown to act as mediators of population density-dependent (quorum-sensing) gene expression in numerous Gram-negative bacteria. Functions associated with AHL include light production in Vibrio fischeri, expression of virulence factors in Pseudomonas aeruginosa, and conjugation in Agrobacterium tumefaciens. In nature, bacteria often grow as surface-adherent biofilm communities. As biofilms typically contain high concentrations of cells, AHL activity and quorum-sensing gene expression have been proposed as essential components of biofilm physiology. However, proof of AHL production within biofilms has heretofore been lacking. In this study we have employed a cross-feeding assay, using A. tumefaciens A136 (traI::lacZ) as an AHL-responsive reporter strain, to show the presence of naturally occurring AHL production in aquatic biofilms growing on submerged stones. AHL was detected in living biofilms and biofilm extracts, but was not present in rocks lacking a biofilm. This represents the first report of AHL activity in naturally occurring biofilms.

Carbon repression in aspergilli

Abstract: Many microorganisms prefer easily metabolizable carbon sources over alternative, less readily metabolized carbon sources. One of the mechanisms to achieve this is repression of the synthesis of enzymes related to catabolism of the alternative carbon sources, i.e. carbon repression. It is now clear that in Aspergillus nidulans and Aspergillus niger the repressor protein CREA plays a major role in carbon repression. CREA inhibits transcription of many target genes by binding to specific sequences in the promoter of these genes. Unfortunately there is little information on other components of the signalling pathway that triggers repression by CREA. In this review we summarize the current understanding of carbon repression in aspergilli.

Identification and roles of non-pathogenic microflora associated with honey bees

Abstract: Microorganisms associated with honey bees, Apis mellifera , and their food include bacteria (Gram-variable pleomorphic bacteria, Bacillus spp., and Enterobacteriaceae), molds (primarily aspergilli and penicillia), and yeasts (mainly Torulopsis spp. ). Eggs, prepupae, pupae, and worker bees emerging from cells as adults are usually free of internal microbes. Microorganisms acquired by larvae through ingestion of contaminated food are usually eliminated through the single defecation that occurs at the end of the feeding period prior to pupation. Emerging adult bees acquire intestinal microflora by food exchange with other bees in the colony and through consumption of pollen. Biochemical contributions of microorganisms to honey bees; the role of microorganisms in the conversion, enhancement, and preservation of pollen stored as bee bread in comb cells; and the production of antimycotic substances by molds and Bacillus spp. from honey bee colonies that are resistant to the fungal disease, chalkbrood, are discussed. An association of Bacillus spp. with bees including honey bees, stingless bees, and solitary bees from tropical and temperate zones appears to have evolved in which female bees inoculate food sources with these bacteria whose chemical products contribute to the elaboration and/or protection from spoilage of food that is stored in the nest. This association is ancient based on results from stingless bees preserved in amber for 25–40 million years. It is concluded that bees, their products, and their associated microorganisms are potential sources of bioactive products including antimicrobial compounds.

Detection and in situ identification of representatives of a widely distributed new bacterial phylum

Abstract: 16S rRNA gene libraries were prepared by polymerase chain reaction amplification and cloning from soil samples taken periodically from a field with genetically modified plants. Sequence analyses of the cloned rDNAs indicated that 140 of them clustered apart from known bacterial phyla. Based on 31 full sequences a new phylum could be defined. It includes Holophaga foetida, ‘Geothrix fermentans’ and Acidobacterium capsulatum as the only cultured species so far. Therefore, this line of descent was named the Holophaga/Acidobacterium phylum. About 50 published partial sequences of cloned rDNAs retrieved from soil, freshwater sediments or activated sludge from different continents indicate the occurrence of further representatives of this phylum. Two specific hybridization probes were constructed for members of one of four subclusters. A careful data analysis revealed the importance and problems of identifying and dealing with artefacts such as chimeric structure when defining new phylogenetic groups based mainly upon cloned amplified rDNAs. For the first time, the presence of bacterial cells representing this group could be shown in soil, sediment, activated sludge and lake snow by in situ hybridization.

A family of Gram-negative bacterial outer membrane factors that function in the export of proteins, carbohydrates, drugs and heavy metals from Gram-negative bacteria

Abstract: Gram-negative bacteria have evolved transport complexes that export macromolecules and toxic substances across the two membranes of the cell envelope in a single energy coupled step. The process requires (1) a cytoplasmic membrane export system, (2) a membrane fusion protein (MFP), and (3) an outer membrane factor (OMF). Families comprising the former two constituents have been described previously. We here present an analysis of the phylogenetic and structural characteristics of the OMF family. Twenty-one members of this family have been identified, and based on available evidence, they function in conjunction with ABC, RND, MFS and/or other types of cytoplasmic transport systems. OMFs exhibit fairly uniform sizes (398–495 residues with two exceptions), and based on computational analyses, they may form β-barrel structures consisting of up to 16 β-strands. Phylogenetic analyses reveal that while the MFPs cluster in accordance with the type of cytoplasmic membrane transport systems with which they function, OMFs do not. We conclude that OMFs probably comprise a family of outer membrane porin-type proteins of uniform structure which did not coevolve with their cognate cytoplasmic membrane transport systems.

Microbial flora in the deepest sea mud of the Mariana Trench.

Abstract: In an attempt to characterize the microbial flora on the deepest sea floor, we isolated thousands of microbes from a mud sample collected from the Mariana Trench. The microbial flora found at a depth of 10 897 m was composed of actinomycetes, fungi, non-extremophilic bacteria, and various extremophilic bacteria such as alkaliphiles, thermophiles, and psychrophiles. Phylogenetic analysis of Mariana isolates based on 16S rDNA sequences revealed that a wide range of taxa were represented.

Secreted enzymes of Aeromonas

Abstract: A hallmark characteristic of species of Aeromonas is their ability to secrete a wide variety of enzymes associated with pathogenicity and environmental adaptability. Among the most intensively studied are beta-lactamases, lipases, hemolytic enterotoxins, proteases, chitinases, nucleases and amylases. Multiple copies of genes encoding each type of enzyme provide additional biological diversity. Except for the chitinases, these multiple copies show little evolutionary relatedness at the DNA level and only limited similarity at the protein level. Indeed a number of the genes, such as nuclease H of A. hydrophila, have no similarity to known prokaryotic or eukaryotic sequences. The challenge is to determine how these genes evolved, where they originated and why Aeromonas possesses them in such abundance and variety.

Microbiological activity of whole and fractionated crude extracts of tea (Camellia sinensis), and of tea components

Abstract: Aqueous extracts of teas (Camellia sinensis) of different types and from various sources inhibited a wide range of pathogenic bacteria, including methicillin-resistant Staphylococcus aureus. Tea extracts were bactericidal to staphylococci and Yersinia enterocolitica at well below 'cup of tea' concentrations. Activity was confined to one of four fractions obtained from a green tea extract by partition chromatography. Testing of pure tea compounds and closely related chemicals suggested that the antibacterial activity of extracts of green tea can be explained by its content of epigallocatechin, epigallocatechin gallate and epicatechin gallate. In black tea extracts, theaflavin and its gallates are additional antibacterially active components.

Microbial ribulose 1,5-bisphosphate carboxylase/oxygenase: a molecule for phylogenetic and enzymological investigation

Abstract: Ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) catalyzes the key reaction of the Calvin reductive pentose phosphate cycle and as such is responsible for life as we know it. This enzyme has been intensively studied for decades. Evidence that RubisCO phylogenies are incongruent with those derived from other macromolecules has been accumulating and recent discoveries have driven home this point. Here we review findings regarding RubisCO phylogeny and discuss these in the context of the important biochemical and structural features of the enzyme. The implications for the engineering of improved RubisCO enzymes are considered.

A broad-host-range mobilizable shuttle vector for the construction of transcriptional fusions to β-galactosidase in Gram-positive bacteria

Abstract: A low-copy-number vector designated pTCV-lac has been constructed to provide a convenient system to analyze regulatory elements in Gram-positive bacteria. The main components of this vector are: (i) the origins of replication of pACYC184 and of the broad-host-range enterococcal plasmid pAMβ1, (ii) erythromycin- and kanamycin-resistance-encoding genes for selection in Gram-negative and Gram-positive bacteria, (iii) the transfer origin of the IncP plasmid RK2, and (iv) a promoterless β-galactosidase-encoding lacZ gene with a Gram-positive ribosome binding site. This 12 kb plasmid is present in Gram-positive hosts in three to five copies per chromosome equivalent and contains three unique cloning sites (EcoRI, SmaI, BamHI) for cloning of DNA inserts upstream of the lacZ gene. Plasmid pTCV-lac and derivatives carrying different promoter fragments have been transferred by conjugation from an Escherichia coli IncP mobilizing donor strain to Bacillus subtilis, Listeria monocytogenes, Enterococcus faecalis, and Streptococcus agalactiae. These plasmids were structurally stable in these hosts and the corresponding promoter activities, quantitated by the determination of the β-galactosidase specific activities, were found to cover at least a 100-fold range in β-galactosidase values. These results indicate that pTCV-lac should be useful for analysis of gene regulation in a wide range of Gram-positive bacteria.

Inhibition of in vitro growth of enteropathogens by new Lactobacillus isolates of human intestinal origin

Abstract: Three human Lactobacillus strains, coded B21060, B21070 and B21190, have recently been isolated. The strains show a series of features (acid and bile resistance, adhesion to various types of mucosal cell) which make them particularly promising for the preparation of probiotic products. In the present study, the ability of the strains to inhibit the growth of pathogens in coculture was investigated. Lactobacilli were incubated simultaneously or after one overnight growth with enterotoxigenic Escherichia coli, Salmonella enteritidis or Vibrio cholerae. After 24 and 48 h, bacterial counts of the pathogens and of the lactobacilli were performed. The results showed that these Lactobacillus strains inhibited the in vitro growth of E. coli and S. enteritidis under both conditions. Moreover, a cumulative effect was observed for mixtures of lactobacilli. In contrast, no significant inhibition of Vibrio cholerae growth was observed, provided that the pH of the medium was kept constant. The presence of the pathogens did not affect the growth of the Lactobacillus strains. Moreover, each of the Lactobacillus strains showed coaggregation ability with two pathogenic E. coli strains, namely ATCC 25922 and ATCC 35401.

Colorimetric protein phosphatase inhibition assay of laboratory strains and natural blooms of cyanobacteria: comparisons with high-performance liquid chromatographic analysis for microcystins

Abstract: Microcystins are cyclic heptapeptide hepatotoxins commonly produced by bloom-forming genera of cyanobacteria. These toxins are potent and specific inhibitors of protein phosphatases 1 and 2A. We have optimised a rapid, simple and sensitive colorimetric protein phosphatase 1 inhibition assay, utilising the activity of protein phosphatase 1 as expressed in a recombinant strain of Escherichia coli, towards the chromogenic substrate, p-nitrophenyl phosphate. A standard curve for the inhibition of protein phosphatase 1 by microcystin-LR was constructed with an IC50 of about 38 ng ml-1 and a limit of detection of 10-20 ng ml-1. Twenty-three laboratory-grown strains and 25 natural bloom samples of cyanobacteria were analysed by high-performance liquid chromatography for microcystins and by the protein phosphatase 1 inhibition assay. Agreement for the microcystin contents of the samples detected by high-performance liquid chromatography and the protein phosphatase 1 inhibition assay showed good correlation (R2 > 0.93, P < 0.0001). The suitability of the colorimetric protein phosphatase 1 inhibition assay as a screen for cyanobacterial microcystins is discussed.

A gene cluster closely related to type II secretion pathway operons of gram-negative bacteria is located on the large plasmid of enterohemorrhagic Escherichia coli O157 strains.

Abstract: Analysis of 14.162 kb of DNA derived from plasmid pO157 of enterohemorrhagic Escherichia coli (EHEC) O157:H7 strain EDL933, extending in the 5′ direction of the recently described EHEC-hly operon, revealed 13 open reading frames (ORF) which showed great similarities to genes of members of the type II pathway secretion systems of Gram-negative bacteria. We named the ORFs etpC to etpO for HEC ype II secretion athway. In addition, an IS911-like insertion element was found to separate the etp genes from the EHEC-hlyC gene. Hybridization experiments with a specific etp probe and various categories of enteric E. coli pathotypes revealed that the etp gene cluster occurred in all 30 EHEC strains of serogroup O157 (100%) tested and is distributed sporadically among other EHEC serogroups (60%). In addition, the etp genes were rarely detected in STEC isolated from bovine feces (10%). Moreover, it was found not to occur in enteropathogenic E. coli, enteroaggregative E. coli, enterotoxigenic E. coli and enteroinvasive E. coli. The results obtained with the etp probe were confirmed by a PCR approach to specifically detect an internal fragment of the etpD gene.

The particulate methane monooxygenase gene pmoA and its use as a functional gene probe for methanotrophs

Abstract: The particulate methane monooxygenase gene pmoA, encoding the 27 kDa polypeptide of the membrane-bound particulate methane monooxygenase, was amplified by PCR from DNA isolated from a blanket peat bog and from enrichment cultures established, from the same environment, using methane as sole carbon and energy source. The resulting 525 bp PCR products were cloned and a representative number of clones were sequenced. Phylogenetic analysis of the derived amino acid sequences of the pmoA clones retrieved directly from environmental DNA samples revealed that they form a distinct cluster within representative PmoA sequences from type II methanotrophs and may originate from a novel group of acidophilic methanotrophs. The study also demonstrated the utility of the pmoA gene as a phylogenetic marker for identifying methanotroph-specific DNA sequences in the environment.

Capsaicin as an inhibitor of the growth of the gastric pathogen Helicobacter pylori

Abstract: Capsaicin, the active ingredient in chili, has been implicated as both a cytoprotective and a detrimental agent to the gastric mucosa. The effect of capsaicin on Helicobacter pylori has not been investigated previously. Therefore, we performed in vitro time- and concentration-dependent studies to examine the growth of H. pylori in the presence of capsaicin. Capsaicin specifically inhibited growth of H. pylori dose-dependently at concentrations greater than 10 μg ml−1 (P<0.05) but did not inhibit the growth of a human fecal commensal Escherichia coli strain. Bactericidal activity was observed within 4 h. Capsaicin continued to exhibit bactericidal activity when incubated at pH values as low as 5.4. Ingestion of chili, therefore, could have a protective effect against H. pylori-associated gastroduodenal disease. This effect deserves further study in animal models.

Molecular mechanisms of azole resistance in fungi

Abstract: This paper reviews the current status of our understanding of azole antifungal resistance mechanisms at the molecular level and explores their implications. Extensive biochemical studies have highlighted a significant diversity in mechanisms conferring resistance to azoles, which include alterations in sterol biosynthesis, target site, uptake and efflux. In stark contrast, few examples document the molecular basis of azole resistance. Those that do refer almost exclusively to mechanisms in laboratory mutants, with the exception of the role of multi-drug resistance proteins in clinical isolates of Candida albicans. It is clear that the technologies required to examine and define azole resistance mechanisms at the molecular level exist, but research appears distinctly lacking in this most important area.

Molecular analysis of cyp51 from fluconazole-resistant Candida albicans strains

Abstract: The target enzyme for fluconazole is sterol 14α-demethylase, a cytochrome P450 encoded by cyp51. One mechanism of fluconazole resistance likely to occur in Candida albicans is through an altered target site. To test this hypothesis DNA sequencing of the cyp51 coding sequence from 19 fluconazole-resistant and 19 fluconazole-sensitive C. albicans was undertaken. A number of point mutations were identified in the resistant isolates which were not present in the sensitive ones: F105L (five), E266D (five), K287R (one), G448G (one), G450E (one), G464S (three) and V488I (one). These alterations are discussed in the light of a molecular model of the enzyme regarding potential roles in resistance. It was also demonstrated that sequence-specific primers can be employed to identify polymorphisms which may be associated with resistance; diagnostic tests for resistant strains will prove of value in combating this serious clinical problem.

Rahnella aquatilis, a bacterium isolated from soybean rhizosphere, can solubilize hydroxyapatite

Abstract: A diazotropic bacterium, ISL19, has been isolated from soybean rhizosphere. ISL19 grows well at 30°C and utilizes diverse carbon sources for its growth. This Gram-negative bacterium is rod-shaped, is about 2–3 μm in size and contains several flagella. On the basis of its cellular fatty acid profile, its carbon utilization pattern, and the nucleotide sequence of a conserved segment of a 16S rRNA gene, ISL19 has been identified as Rahnella aquatilis. This bacterium exhibited a strong ability to solubilize hydroxyapatite and dicalcium phosphate in the external culture medium. The solubilization of hydroxyapatite was associated with a drop in the pH of the culture medium. The change in the pH value showed an inverse correlation with the soluble phosphorus concentration. Analysis of the culture medium by high pressure liquid chromatography identified gluconic acid as the main organic acid released by R. aquatilis.

Fabatins: new antimicrobial plant peptides

Abstract: Two new antimicrobial peptides related to the γ-thionine family have been isolated by acid extraction from the broad bean Vicia faba. The extract was separated by ion exchange chromatography, and a fraction showing antibacterial activity was further purified by reverse-phase HPLC. Material from a single HPLC peak was sequenced and revealed the presence of two peptides differing by one amino acid. The peptides were named fabatins. They are 47 amino acids long, have an overall positive charge and contain 8 cysteines that probably form 4 disulfide bridges characteristic of the γ-thionins. Fabatins were active against both Gram-negative and Gram-positive bacteria, but were inactive against the yeasts Saccharomyces cerevisiae and Candida albicans.

Molecular aspects of the E. coli nucleoid protein, H-NS: a central controller of gene regulatory networks

Abstract: The nucleoid-associated protein H-NS has a central role in the structuring and control of the enteric bacterial chromosome. This protein has been demonstrated to contribute to the regulation of expression for approximately thirty genes. In this article, the molecular aspects of H-NS structure and function are briefly reviewed. H-NS contains at least two independent structural domains: a C-terminal domain, involved in the DNA-protein interactions, and a N-terminal domain, likely involved in protein–protein interactions. Recent reports have revealed that H-NS is a key factor in a multi-component gene regulatory system. Factors have now been discovered which can backup or antagonise H-NS action at certain promoters. These recent findings are summarised and discussed in relationship to the role of H-NS in DNA packaging and nucleoid structure.

Blue and yellow laccases of ligninolytic fungi

Abstract: Extracellular laccases from submerged cultures of Coriolus versicolor BKM F-116, Panus tigrinus 8/18, Phlebia radiata 79 (ATCC 64658), Phlebia tremellosa 77-51 and from cultures of Pa. tigrinus 8/18, Ph. radiata 79 and Agaricus bisporus D-649 grown on wheat straw (solid-state fermentation) were purified. All enzymes from submerged cultures had a blue colour and characteristic absorption and EPR spectra. Laccases from the solid-state cultures were yellow-brown and had no typical blue oxidase spectra and also showed atypical EPR spectra. Comparison of N-terminal amino acid sequences of purified laccases showed high homology between blue and yellow-brown laccase forms. Formation of yellow laccases as a result of binding of lignin-derived molecules by enzyme protein is proposed.

Melanization affects susceptibility of Cryptococcus neoformans to heat and cold.

Abstract: Cryptococcus neoformans can synthesize melanin from a variety of substrates, including L-dopa (l-3,4-dihydroxyphenylalanine). Growth in minimal medium with L-dopa resulted in progressive accumulation of melanin in stationary phase cells. Melanized and non-melanized yeast cells were exposed to heat (42–47°C) and cold (−20°C), and the percentage of survival determined. Melanized cells were less susceptible to heat than non-melanized cells of the same age. Melanized cells from early stationary phase cultures were less susceptible to cold than non-melanized cells of the same age. However, melanized cells from late stationary phase cultures were more susceptible to cold than non-melanized cells of the same age. There was no statistical difference in susceptibility to heat and cold between melanin-deficient cells grown with and without L-dopa. These results suggest a role for melanin in protection against heat and cold.

Antigen 43, a phase‐variable bipartite outer membrane protein, determines colony morphology and autoaggregation in Escherichia coli K‐12

Abstract: The product controlling colony form variation and autoaggregation in Escherichia coli K-12 (the flu gene product) has been identified as the phase-variable, bipartite, outer membrane protein, termed antigen 43 (Ag43). Identification is based: (i) on complete correlation in authentic flu variants between colony morphology/autoaggregation and Ag43 expression as determined by colony and Western immunoblotting and immunofluorescence microscopy; and (ii) on the use of a specific probe to map the gene encoding Ag43 to a position (min 43) on the E. coli chromosome previously established for flu.

Mutants of Saccharomyces cerevisiae defective in vacuolar function confirm a role for the vacuole in toxic metal ion detoxification

Abstract: To directly define vacuolar role(s) in metal detoxification, we have examined the responses of vacuole-deficient mutants of Saccharomyces cerevisiae to several potentially toxic metals known to be mainly detoxified in the cytosol (Cu, Cd) or the vacuole (Co, Mn, Ni, Zn). Three mutants, deficient in targeting of vacuolar proteins, were used with JSR18Δ1 being devoid of any vacuole-like structure while ScVatB and ScVatC were deficient in specific protein subunits of the V-ATPase. The results obtained show that the absence of a vacuole or a functional vacuolar H+-ATPase was associated with increased sensitivity and a largely decreased capacity of the vacuole-deficient strains to accumulate Zn, Mn, Co and Ni, confirming an essential role for the vacuole in detoxification of these metals. In addition, the lack of vacuolar involvement in detoxification of Cu and Cd was confirmed since these metals did not exhibit increased toxicity towards the vacuolar mutants nor were there significant differences in Cu or Cd accumulation between parental and mutant strains.

Bacterial degradation of natural rubber: a privilege of actinomycetes?

Abstract: Using natural rubber latex as the sole source of carbon and energy 50 rubber-degrading bacteria were isolated. Out of those 50 isolates, 33 were identified as Streptomyces species and 8 as Micromonospora species. Screening of 1220 bacteria obtained from different culture collections revealed 46 additional rubber-degrading bacteria ( Streptomyces 31 strains, Micromonospora 5, Actinoplanes 3, Nocardia 2, Dactylosporangium 1, Actinomadura 1, unidentified 3). All rubber-degrading isolates were identified as members of the actinomycetes, a large group of mycelium-forming Gram-positive bacteria. Interestingly no Gram-negative bacterium could be isolated. In most strains expression of extracellular rubber-degrading enzymes was repressed by glucose and/or succinate. The reduction of the average molecular mass of solution-cast films of natural rubber from 640 000 to 25 000 in liquid culture upon bacterial growth indicates the participation of an endo -cleavage mechanism of degradation.