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Biomedical subjects

M G Bramucci

Publications and source records attributed to M G Bramucci.

At least 19 recordsLinked to original sources
PubMed2002

Pure bacterial isolates that convert p-xylene to terephthalic acid.

Bacteria that grow on p-xylene, p-toluic acid, and terephthalic acid (TPA) were isolated from a wastewater bioreactor that is used to treat a waste stream that contains all three of these compounds. Although previously described aerobic bacteria degrade p-xylene by initially oxidizing a single methyl group to form p-toluic acid and then cleaving the aromatic ring, some of the bacteria isolated during this study transformed p-xylene by oxidizing both methyl groups to produce TPA.

Bioreactors↗
PubMed2000

Industrial wastewater bioreactors: sources of novel microorganisms for biotechnology.

Microorganisms exist in nature as members of complex, mixed communities. The microbial communities in industrial wastewater bioreactors can be used as model systems to study the evolution of new metabolic pathways in natural ecosystems. The evolution of microbial metabolic capability in these bioreactors is presumably analogous to phenomena that occur in natural ecosystems. The microorganisms in these bioreactors compete for different carbon sources and constantly have to evolve new metabolic capabilities for survival. Thus, industrial bioreactors should be a rich source of novel biocatalysts.

Bacteria↗
PubMed1996

Direct selection of cloned DNA in Bacillus subtilis based on sucrose-induced lethality.

Expression of the Bacillus subtilis or Bacillus amyloliquefaciens sacB gene in the presence of sucrose is lethal for a variety of bacteria. Sucrose-induced lethality can be used to select for inactivation of sacB by insertion of heterologous DNA in sensitive bacteria. This procedure has not been applicable to B. subtilis heretofore because expression of wild-type sacB is not detrimental to B. subtilis. The W29 mutation in the B. amyloliquefaciens sacB gene interferes with processing of the levansucrase signal peptide. The W29 mutation does not affect growth of B. subtilis in media lacking sucrose. However, this mutation inhibited growth of B. subtilis in media containing sucrose. Inactivation of the fructose polymerase activity encoded by sacB indicated that levan production was essential for sucrose-induced lethality. As a result, it was possible to select for cloned DNA in B. subtilis by insertional inactivation of the mutant sacB gene located on a multicopy plasmid vector in medium containing sucrose.

Bacillus↗
PubMed1991

The Bacillus subtilis spo0J gene: evidence for involvement in catabolite repression of sporulation.

Previous observations concerning the ability of the Bacillus subtilis bacteriophages SP10 and PMB12 to suppress mutations in spo0J and to make wild-type sporulation catabolite resistant suggested that spo0J had a role in catabolite repression of sporulation. This suggestion was supported in the present report by the ability of the catabolite-resistant sporulation mutation crsF4 to suppress a Tn917 insertion mutation of the B. subtilis spo0J locus (spo0J::Tn917 omega HU261) in medium without glucose. Although crsF4 and SP10 made wild-type B. subtilis sporulation catabolite resistant, neither crsF4 nor SP10 caused a mutant with spo0J::Tn917 omega HU261 to sporulate in medium with glucose. Sequencing the spo0J locus revealed an open reading frame that was 179 codons in length. Disruption of the open reading frame resulted in a sporulation-negative (Spo-) phenotype that was similar to those of other spo0J mutations. Analysis of the deduced amino acid sequence of the spo0J locus indicated that the spo0J gene product contains an alpha-helix-turn-alpha-helix unit similar to the motif found in lambda Cro-like DNA-binding proteins.

Amino Acid Sequence↗
PubMed1990

Bacteriophage-enhanced sporulation: comparison of spore-converting bacteriophages PMB12 and SP10.

The previously characterized bacteriophage SP10 enhanced the frequency of wild-type sporulation by Bacillus subtilis W23 and 3-13. Comparison of SP10 with the spore-converting bacteriophage PMB12 indicated that both bacteriophages significantly increased the sporulation frequency of an oligosporogenic mutant that contained spo0J::Tn917 omega HU261. SP10 and PMB12 caused wild-type bacteria to sporulate in a liquid medium that initially contained enough glucose to inhibit the sporulation and expression of alpha-amylase by uninfected bacteria. SP10 also induced the expression of alpha-amylase in the presence of glucose, whereas PMB12 had no detectable effect. These observations were consistent with the conclusion that SP10 is a spore-converting bacteriophage and that SP10 and PMB12 relieve glucose-mediated catabolite repression of sporulation by different mechanisms.

Bacillus subtilis↗
PubMed1982

Mapping of a gene in Vibrio cholerae that determines the antigenic structure of cholera toxin.

In Ouchterlony-type immunodiffusion gels, the cholera toxin produced by the classical Vibrio cholerae strain 569B was indistinguishable from the cholera toxin of the Eltor strain RJ1. However, the cholera toxin produced by both of these strains was incompletely cross-reactive with the cholera toxin produced by the Eltor strain 3083-2. The allele of the gene responsible for the 569B and RJ1 type of toxin was designated vct-1, and the allele responsible for the 3083-2 type of toxin was designated vct-2. The vct-1 allele was transferred from RJ1 donors to 3083-2 recipients by conjugation. The vct locus was found to be between met and trp on the V. cholerae linkage map.

Alleles↗
PubMed1981

Isolation and characterization of hypertoxinogenic (htx) mutants of Escherichia coli KL320(pCG86).

The structural genes for heat-labile enterotoxin (LT) are present on plasmid pCG86. Escherichia coli KL320(pCG86), LT was found to be cell associated. LT was present as a soluble protein in sonic lysates of KL320(pCG86). Thirty-one mutants of KL320(pCG86) that produced increased amounts of extracellular LT were isolated. These hypertoxinogenic (htx) mutants were assigned to four phenotypically distinct classes based on the amounts of cell-associated and extracellular LT in early-stationary-phase cultures. Type 1 and type 2 htx mutants produced significantly increased amounts of cell-associated LT. Type 3 and type 4 htx mutants produced normal or decreased amounts of cell-associated LT was similar to that of the wild type. In the mutants of types 1, 3, and 4, the ratios of extracellular to cell-associated LT were higher than that of the wild type and were characteristic for each strain. Cell lysis or leakage of macromolecular cytoplasmic constituents appeared to be significant for release of LT by mutants of types 1, 3, and 4, because supernatants from cultures of these mutants also contained increased amounts of protein and of the cytoplasmic enzyme glucose 6-phosphate dehydrogenase. In all four representative htx mutants, the hypertoxinogenic phenotypes were dependent on chromosomal mutations. The resident pCG86 plasmids were eliminated from the htx mutants of types 2 and 3. After wild-type plasmid pCG86 was introduced into the cured strains by conjugation, their hypertoxinogenic phenotypes were restored. We conclude that chromosomal loci in E. coli KL320 are important in regulating expression of the LT structural genes of plasmid pCG86.

Bacterial Toxins↗
PubMed1981

Analysis of Bacillus subtilis sporulation with spore-converting bacteriophage PMB12.

Previous observations concerning the ability of the spore-converting bacteriophage PMB12 to cause sporulation in certain sporulation-deficient mutants of Bacillus subtilis 168 were extended to include a spoOK mutant and a mutant temperature sensitive for sporulation due to a ribosomal mutation. Mutants of PMB12 that were unable to induce sporulation in the spoOK mutant were isolated to determine whether PMB12-encoded products had to affect the sporulation-specific functions of both the transcription and the translation systems of B. subtilis to induce sporulation. A complementation assay for spore conversion was used to assign the spore conversion-negative PMB12 mutants to four groups. One group of mutants repressed the ability of wild-type PMB12 to induce sporulation. None of the spore conversion-negative PMB12 mutants could induce significant levels of sporulation in B. subtilis mutants that were temperature sensitive for sporulation due to mutations in the beta subunit of ribonucleic acid polymerase or the 30S ribosomal subunit. Our data suggest that PMB12 may have at least three genes for spore conversion. The products of these genes apparently interact with a host cell pathway that is expressed during the earliest stage of sporulation and is not dependent for expression upon sporulation-specific functions of the host cell's transcription and translation systems.

Bacillus subtilis↗
PubMed1977

Host function specified by Bacillus pumilus plasmid pPL7065.

Plasmid pPL7065 ( approximately 4.7 x 10(6) daltons; approximately 20 copies per chromosome) determines the production of, and immunity or resistance to, a killing activity in strains of Bacillus pumilus. Plasmid pPL7065 is compatible with plasmid pPL576 ( approximately 28 x 10(6) daltons; approximately 2 copies per chromosome).

Bacillus↗
PubMed1977

Selective plasmid transduction in Bacillus pumilus.

The inducible temperate bacteriophage phi75 and a clear-plaque-forming variant, phi75C1, mediated transduction of a 4.4 X 10(6)-dalton multicopy Bacillus pumilus plasmid, pPL10, at frequencies of 10(-5) to 10(-6) transductants per plaque-forming unit. phi75- and phi75C1-mediated transduction of several chromosome markers tested did not occur at a detectable frequency. phi75-mediated plasmid transducing activity resides in particles that are similar to infectious particles in sedimentation velocity and buoyant density.

Bacillus↗
PubMed1977

Bacteriophage conversion of spore-negative mutants to spore-positive in Bacillus pumilus.

A pseudolysogenic phage, PMB1, was isolated from soil on the basis of its ability to increase the sporulation frequency of the oligosporogenic Bacillus pumilus strain NRS 576 (sporulation frequency, less than 1%). Several spore-negative mutants (sporulation frequency, less than 10-8) derived from strain NRS 576, which were converted to spore positive by infection with PMB1, were subsequently identified. PMB1 repeatedly grown on a given spore-negative mutant (e.g., GW2) converted GW2 cells to spore positive. Each plaque-forming unit initiated the conversion of a spore-positive clone in semisolid agar overlays. GW2 cells remained spore positive as long as they maintained PMB1. Return of PMB1-converted cells to the orginal spore-negative phenotype correlated with loss of PMB1. In liquid media, PMB1 infection increased the sporulation frequency of mutant GW2 over 106-fold. More than half of the spore-negative mutants we isolated from strain NRS 576 were converted to spore positive by PMB1 infection. PMB1-induced spores of the spore-negative mutant GW2 were somewhat more heat sensitive than uninfected or PMB1-infected spores of the spore positive parent of GW2. PMB1-induced spores of GW2 do not differ from wild-type spores in morphology by phase-contrast microscopy, dipicolinic acid content, or rate of sedimentation through Renografin gradients.

Bacillus↗
PubMed1977

Bacteriophage PMB12 conversion of the sporulation defect in RNA polymerase mutants of Bacillus subtilis.

The pseudotemperate phage PMB12 was isolated from soil on the basis of its ability to enhance the rate of sporulation of Bacillus subtilis 168. PMB12 was subsequently shown to convert the sporulation defect in two genetically distinct classes of sporulation mutants. One class includes those rifampin-resistant mutants that are also spore-negative (mutated at the rif locus). The other class includes a strain carrying the sporulation mutation spoCM-1. The spoCM-1 mutation is linked to cysA15 by PBS1 transduction but is distinct from the rif locus. Several other sporulation mutants were not converted by PMB12. PMB12 is related to phage PBS1. However, PBS1 did not convert the above sporulation mutants. The replication of PBS2, a clear-plaquing derivative of PBS1, is rifampin insensitive, apparently due to a phage-induced rifampin-insensitive RNA polymerase. PMB12 replication is also rifampin insensitive.

Bacillus subtilis↗