TitleAbstractYear(sorted ascending)
transposon tn21, flagship of the floating genome.the transposon tn21 and a group of closely related transposons (the tn21 family) are involved in the global dissemination of antibiotic resistance determinants in gram-negative facultative bacteria. the molecular basis for their involvement is carriage by the tn21 family of a mobile dna element (the integron) encoding a site-specific system for the acquisition of multiple antibiotic resistance genes. the paradigm example, tn21, also carries genes for its own transposition and a mercury resistanc ...199910477306
principles and applications of methods for dna-based typing of microbial organisms. 199910325304
rapid identification of up to three candida species in a single reaction tube by a 5' exonuclease assay using fluorescent dna probes.we used fungus-specific pcr primers and species-specific dna probes to detect up to three candida species in a single reaction tube by exploiting the 5' to 3' exonuclease activity of taq dna polymerase. probes to the internal transcribed spacer region of the rrna gene were labeled at the 5' end with one of three fluorescent reporter dyes, 6-carboxy-fluorescein (fam), tetrachloro-6-carboxy-fluorescein (tet), or hexachloro-6-carboxy-fluorescein (hex), and at the 3' end with a quencher dye, 6-carbo ...19999854084
molecular epidemiology of ceftazidime-resistant gram-negative bacilli on inanimate surfaces and their role in cross-transmission during nonoutbreak periods.we described the molecular epidemiology of expanded-spectrum cephalosporin-resistant gram-negative bacilli (rgn) recovered from inanimate surfaces. rgn were isolated from 9% of environmental cultures. numerous species, each with multiple unique strains, were recovered. epidemiological links between environmental, personnel, and patient strains suggested the exogenous acquisition of rgn from the hospital environment.199910449510
polypeptide requirement of multicomponent monooxygenase dsoabcdef for dimethyl sulfide oxidizing our previous study, the multicomponent monooxygenase dsoabcdef in acinetobacter sp. strain 20b was cloned based on its ability to oxidize dimethyl sulfide (dms) to dimethyl sulfoxide (dmso) in e. coli cells, which had high sequence similarity with phenol hydroxylase mopklmnop in acinetobacter calcoaceticus ncib8250, dmpklmnop in pseudomonas sp. cf600 and some other multicomponent monooxygenases. in this study, dsob, c, d, e, and f were found to be needed for dms-oxidizing activity in polypept ...199926300166
nature, nomenclature and taxonomy of obligate methanol utilizing a screening program, a number of different bacterial strains with the ability to utilize methanol as a sole carbon and energy source were isolated and described. they are well known methanol utilizing genera pseudomonas, klebsiella, micrococcus, methylomonas or, on the contrary, the new, unknown genera and species of methylotrophic bacteria. in the last category, acinetobacter and alcaligenes are the new reported genera of organisms able to use methanol as a sole carbon and energy source. the ...199911845445
a protein residing at the subunit interface of the bacterial ribosome.surface labeling of escherichia coli ribosomes with the use of the tritium bombardment technique has revealed a minor unidentified ribosome-bound protein (spot y) that is hidden in the 70s ribosome and becomes highly labeled on dissociation of the 70s ribosome into subunits. in the present work, the n-terminal sequence of the protein y was determined and its gene was identified as yfia, an orf located upstream the phe operon of e. coli. this 12.7-kda protein was isolated and characterized. an af ...199910535924
allylic or benzylic stabilization is essential for catalysis by bacterial benzyl alcohol dehydrogenases.benzyl alcohol dehydrogenase from acinetobacter calcoaceticus (ac-badh) and tol plasmid-encoded benzyl alcohol dehydrogenase from pseudomonas putida (tol-badh) have previously been shown to oxidize a variety of aromatic alcohols but not aliphatic substrates. here, we have expressed the genes for ac-badh and tol-badh in escherichia coli, purified the resulting over-expressed enzymes, and shown that each is an effective catalyst of both benzylic and allylic alcohol oxidation, but not of oxidation ...199910334943
rrna probe-based cell fishing of bacteria.we have developed a new, cultivation-independent, fast and flexible method for the rrna-targeted probe-based enrichment of bacteria. the target cells were labelled by in situ hybridization with biotinylated polyribonucleotide probes. these probes were generated by in vitro transcription of amplified rdna of a variable region in domain iii of the 23s rrna molecules. the probes were about 300 nucleotides in length and were labelled by incorporation of biotin-utp during the transcription. probes we ...199911207744
molecular screening for alkane hydroxylase genes in gram-negative and gram-positive strains.we have developed highly degenerate oligonucleotides for polymerase chain reaction (pcr) amplification of genes related to the pseudomonas oleovorans gpo1 and acinetobacter sp. adp1 alkane hydroxylases, based on a number of highly conserved sequence motifs. in all gram-negative and in two out of three gram-positive strains able to grow on medium- (c6-c11) or long-chain n-alkanes (c12-c16), pcr products of the expected size were obtained. the pcr fragments were cloned and sequenced and found to e ...199911207749
genotypic and phenotypic similarity of multiresistant acinetobacter baumannii isolates in the czech republic.the diversity of 103 clinical isolates of the acinetobacter calcoaceticus-acinetobacter baumannii complex obtained between 1991 and 1997 from 17 czech hospitals was studied by ribotyping, biotyping, plasmid profiling and antibiotic susceptibility testing. according to the ecori ribotypes, all but one of these isolates were identified to the dna group level: 77 isolates were allocated to dna group 2 (a. baumannii), 14 to dna group 3, 10 to dna group 13 sensu tjernberg and ursing and one to dna gr ...199910334596
distribution of acinetobacter species on skin of healthy humans.the distribution of the 19 currently known genospecies of acinetobacter on human skin, i.e. forehead, forearm and toe webs, was determined. three selective media were compared for their specificity for all genospecies of acinetobacter. a minimal-salts agar supplemented with 1% acetate proved to be more efficient than the leeds medium for the isolation of most genospecies in mixed culture with other bacterial species. acinetobacter isolates were provisionally identified using biochemical tests an ...199910357050
epidemiological study of an acinetobacter baumannii outbreak by using a combination of antibiotyping and ribotyping.from june to november 1994 (period 1) and from february to june 1995 (period 2), multiresistant acinetobacter baumannii strains were isolated in intensive care units and surgical wards of the amiens teaching hospital center (amiens, france). eighteen isolates were obtained from 17 (1%) of 1,706 patients admitted during both of these periods, giving an incidence rate of nosocomial infection per 1,000 patient days of 0.6%. of 17 infected patients, 9 had pneumonia, 3 had urinary tract infection, 2 ...199910364581
[trends in resistant bacteria isolated from a tube smear in intubated patients in intensive care].long standing antibiotics therapy has resulted in growing bacteria resistance. we took a tube smear and prepared culture with antibiogram from the fifty intubated patients in the intensive care unit in the war period. gram-negative germs were the dominant ones in total sum, and among them the acinetobacter calcoaceticus (no 21), pseudomonas aeruginosa (no 18), klebsiella pneumoniae (no 18), were isolated most frequently. pseudomonas aeruginosa showed high resistance to gentamicin (64%), amikacin ...199910386042
cell surface analysis techniques: what do cell preparation protocols do to cell surface properties?cell surface analysis often requires manipulation of cells prior to examination. the most commonly employed procedures are centrifugation at different speeds, changes of media during washing or final resuspension, desiccation (either air drying for contact angle measurements or freeze-drying for sensitive spectroscopic analysis, such as x-ray photoelectron spectroscopy), and contact with hydrocarbon (hydrophobicity assays). the effects of these procedures on electrophoretic mobility, adhesion to ...199910388679
physiological adaptations involved in alkane assimilation at a low temperature by rhodococcus sp. strain q15.we examined physiological adaptations which allow the psychrotroph rhodococcus sp. strain q15 to assimilate alkanes at a low temperature (alkanes are contaminants which are generally insoluble and/or solid at low temperatures). during growth at 5 degrees c on hexadecane or diesel fuel, strain q15 produced a cell surface-associated biosurfactant(s) and, compared to glucose-acetate-grown cells, exhibited increased cell surface hydrophobicity. a transmission electron microscopy examination of strai ...199910388690
structure of the soluble methane monooxygenase regulatory protein b.the soluble methane monooxygenase (smmo; ec from the pseudothermophile methylococcus capsulatus (bath) is a three-component enzyme system that catalyzes the selective oxidation of methane to methanol. we have used nmr spectroscopy to produce a highly refined structure of mmob, the 16-kda regulatory protein of this system. this structure has a unique and intricate fold containing seven beta-strands forming two beta-sheets oriented perpendicular to each other and bridged by three alpha ...199910393915
specificity of the lipase-specific foldases of gram-negative bacteria and the role of the membrane anchor.folding of lipases that are secreted by pseudomonads and other gram-negative bacteria via the type ii secretion pathway is facilitated by dedicated chaperones, called lipase-specific foldases (lifs). lifs are membrane-anchored proteins with a large periplasmic domain. the functional interaction between the lif and its cognate lipase is specific, since the pseudomonas aeruginosa lif was found not to substitute for lifs from burkholderia glumae or acinetobacter calcoaceticus. however, the p. aerug ...199910394914
site-directed mutagenesis study on the thermal stability of a chimeric pqq glucose dehydrogenase and its structural interpretation.we have previously reported that a chimeric pyrroloquinoline quinone (pqq) glucose dehydrogenase (gdh), e97a3, which was made up of 97% of escherichia coli pqqgdh sequence and 3% of acinetobacter calcoaceticus pqqgdh, showed increased thermal stability compared with both parental enzymes. site-directed mutagenesis studies were carried out in order to investigate the role of amino-acid substitution at the c-terminal region, ser771, of a chimeric pqqgdhs on their thermal stability. a series of ser ...199910399272
the genes ruba and rubb for alkane degradation in acinetobacter sp. strain adp1 are in an operon with estb, encoding an esterase, and oxyr.alkanes are oxidized in acinetobacter sp. strain adp1 by a three-component alkane monooxygenase, composed of alkane hydroxylase, rubredoxin, and rubredoxin reductase. ruba and rubb encode rubredoxin and a nad(p)h-dependent rubredoxin reductase. we demonstrate here that single base pair substitutions in ruba or rubb lead to defects in alkane degradation, showing that both genes are essential for alkane utilization. differences in the degradation capacity for hexadecane and dodecane in these mutan ...199910400587
areabc genes determine the catabolism of aryl esters in acinetobacter sp. strain adp1.acinetobacter sp. strain adp1 is able to grow on a range of esters of aromatic alcohols, converting them to the corresponding aromatic carboxylic acids by the sequential action of three inducible enzymes: an area-encoded esterase, an areb-encoded benzyl alcohol dehydrogenase, and an arec-encoded benzaldehyde dehydrogenase. the are genes, adjacent to each other on the chromosome and transcribed in the order arecba, were located 3.5 kbp upstream of benk. benk, encoding a permease implicated in ben ...199910419955
nosocomial meningitis in children after ventriculoperitoneal shunt insertion.this study reviews 33 cases of ventriculoperitoneal shunt (vps) meningitis among 415 children after 540 shunt insertions within 8 y, in 9 paediatric intensive care units from 5 centres in slovakia. the incidence of vps meningitis was 6.3% per insertion. the most common organisms isolated from cerebrospinal fluid (csf) and shunt were coagulase-negative staphylococci (52.8%), followed by staphylococcus aureus (13.1%) and pseudomonas aeruginosa (7.5%). seven of 15 assessed risk factors were signifi ...199910426184
a functional 4-hydroxysalicylate/hydroxyquinol degradative pathway gene cluster is linked to the initial dibenzo-p-dioxin pathway genes in sphingomonas sp. strain rw1.the bacterium sphingomonas sp. strain rw1 is able to use dibenzo-p-dioxin, dibenzofuran, and several hydroxylated derivatives as sole sources of carbon and energy. we have determined and analyzed the nucleic acid sequence of a 9,997-bp hindiii fragment downstream of cistrons dxna1a2, which encode the dioxygenase component of the initial dioxygenase system of the corresponding catabolic pathways. this fragment contains 10 colinear open reading frames (orfs), apparently organized in one compact op ...199910348858
proton-nuclear magnetic resonance analyses of the substrate specificity of a beta-ketolase from pseudomonas putida, acetopyruvate hydrolase.a revised purification of acetopyruvate hydrolase from orcinol-grown pseudomonas putida orc is described. this carbon-carbon bond hydrolase, which is the last inducible enzyme of the orcinol catabolic pathway, is monomeric with a molecular size of approximately 38 kda; it hydrolyzes acetopyruvate to equimolar quantities of acetate and pyruvate. we have previously described the aqueous-solution structures of acetopyruvate at ph 7.5 and several synthesized analogues by (1)h-nuclear magnetic resona ...199910438778
fluorescence in situ hybridization assay using peptide nucleic acid probes for differentiation between tuberculous and nontuberculous mycobacterium species in smears of mycobacterium cultures.tb pna fish is a new fluorescence in situ hybridization (fish) method using peptide nucleic acid (pna) probes for differentiation between species of the mycobacterium tuberculosis complex (mtc) and nontuberculous mycobacteria (ntm) in acid-fast bacillus-positive (afb+) cultures is described. the test is based on fluorescein-labelled pna probes that target the rrna of mtc or ntm species applied to smears of afb+ cultures for microscopic examination. parallel testing with the two probes serves as ...199910449448
skin carriage of acinetobacters in hong kong.we studied the carriage of acinetobacter spp. at five superficial sites in 79 patients from two hospitals, in 133 healthy controls from the community (medical students and new nurses), and in 198 student nurses in different classes. a total of 431 isolates from 364 positive sites of 201 subjects and 124 blood culture isolates (1997 to 1998) were genospeciated by amplified ribosomal dna restriction analysis. genospecies 3 was the most common species. the carriage rate of student nurses (42 of 131 ...199910449482
legionella pneumophila invasion of mrc-5 cells induces tyrosine protein phosphorylation.after uptake and intracellular multiplication of legionella pneumophila in mrc-5 lung fibroblasts, important cytoskeletal filament structures, like actin, tubulin, or vimentin, and a cell membrane-associated fibronectin were rearranged during early infection, resulting in a loss of cell adhesion and collapse of the cytoskeleton. dysregulation of the cellular phosphorylation and dephosphorylation cascade may contribute to the observed changes and may support intracellular survival and multiplicat ...199910456891
the physiological contribution of acinetobacter pcak, a transport system that acts upon protocatechuate, can be masked by the overlapping specificity of vank.vank is the fourth member of the ubiquitous major facilitator superfamily of transport proteins to be identified that, together with pcak, benk, and muck, contributes to aromatic catabolism in acinetobacter sp. strain adp1. vank and pcak have overlapping specificity for p-hydroxybenzoate and, most clearly, for protocatechuate: inactivation of both proteins severely impairs growth with protocatechuate, and the activity of either protein alone can mask the phenotype associated with inactivation of ...199910348864
pathophysiology of surgical site infection in total hip arthroplasty.this article is a case report of a 69-year-old man who underwent a right total hip replacement procedure and developed a surgical site infection. areas of concern in prevention and treatment of hip arthroplasty infection are presented, focusing on the pathophysiologic process involved. a review of the patient risk factors and the pathophysiologic action potentiating risk for infection include host immunity, nutritional status, diabetes, age, use of steroids or immunosuppressive drugs, rheumatoid ...199910586159
polyphosphate kinase of acinetobacter sp. strain adp1: purification and characterization of the enzyme and its role during changes in extracellular phosphate levels.polyphosphate (polyp) is a ubiquitous biopolymer whose function and metabolism are incompletely understood. the polyphosphate kinase (ppk) of acinetobacter sp. strain adp1, an organism that accumulates large amounts of polyp, was purified to homogeneity and characterized. this enzyme, which adds the terminal phosphate from atp to a growing chain of polyp, is a 79-kda monomer. ppk is sensitive to magnesium concentrations, and optimum activity occurs in the presence of 3 mm mgcl(2). the optimum ph ...199910473375
bacterial growth state distinguished by single-cell protein profiling: does chlorination kill coliforms in municipal effluent?municipal effluent is the largest reservoir of human enteric bacteria. its public health significance, however, depends upon the physiological status of the wastewater bacterial community. a novel immunofluorescence assay was developed and used to examine the bacterial growth state during wastewater disinfection. quantitative levels of three highly conserved cytosolic proteins (dnak, dps, and fis) were determined by using enterobacterium-specific antibody fluorochrome-coupled probes. enterobacte ...199910473432
the 1.7 a crystal structure of the apo form of the soluble quinoprotein glucose dehydrogenase from acinetobacter calcoaceticus reveals a novel internal conserved sequence repeat.the crystal structure of a dimeric apo form of the soluble quinoprotein glucose dehydrogenase (s-gdh) from acinetobacter calcoaceticus has been solved by multiple isomorphous replacement followed by density modification, and was subsequently refined at 1. 72 a resolution to a final crystallographic r-factor of 16.5% and free r-factor of 20.8% [corrected]. the s-gdh monomer has a beta-propeller fold consisting of six four-stranded anti-parallel beta-sheets aligned around a pseudo 6-fold symmetry ...199910366508
identification and characterization of come and comf, two novel pilin-like competence factors involved in natural transformation of acinetobacter sp. strain bd413.although the high level of competence for natural transformation of acinetobacter sp. strain bd413 has been the subject of numerous studies, only two competence genes, comc and comp, have been identified to date. by chromosomal walking analysis we found two overlapping open reading frames, designated come and comf, starting 61 bp downstream of comc. come and comf are expressed as stable proteins in escherichia coli, thus proving that they are indeed coding regions, but expression was successful ...199910508090
a gene involved in quinate metabolism is specific to one dna homology group of xanthomonas campestris.a gene involved in quinate metabolism was cloned from xanthomonas campestris pv. juglandis strain c5. the gene, quma, located on a 4. 2-kb kpni-ecorv fragment in plasmid pqm38, conferred quinate metabolic activity to x. c. pv. celebensis. tn3-spice insertional analyses further located the quma gene on a region of about 3.0 kb within pqm38. nucleotide sequencing of this 3.0-kb fragment reveals that the coding region of quma is 2373 bp, the deduced amino acid sequence of which closely resembles a ...199910594704
characterization of the phthalate permease ophd from burkholderia cepacia atcc 17616.the ophd gene, encoding a permease for phthalate transport, was cloned from burkholderia cepacia atcc 17616. expression of the gene in escherichia coli results in the ability to transport phthalate rapidly into the cell. uptake inhibition experiments show that 4-hydroxyphthalate, 4-chlorophthalate, 4-methylphthalate, and cinchomeronate compete for the phthalate permease. an ophd knockout mutant of 17616 grows slightly more slowly on phthalate but is still able to take up phthalate at rates equiv ...199910498738
isolation and characterization of two cryptic plasmids in the ammonia-oxidizing bacterium nitrosomonas sp. strain eni-11.two plasmids were discovered in the ammonia-oxidizing bacterium nitrosomonas sp. strain eni-11, which was isolated from activated sludge. the plasmids, designated pays and payl, were relatively small, being approximately 1.9 kb long. they were cryptic plasmids, having no detectable plasmid-linked antibiotic resistance or heavy metal resistance markers. the complete nucleotide sequences of pays and payl were determined, and their physical maps were constructed. there existed two major open readin ...199910348848
identification of acinetobacter baumannii strains with monoclonal antibodies against the o antigens of their lipopolysaccharides.despite the emergence of acinetobacter baumannii strains as nosocomial pathogens, simple methods for their phenotypic identification are still unavailable. murine monoclonal antibodies specific for the o-polysaccharide moiety of the lipopolysaccharide (lps) of two a. baumannii strains were obtained after immunization with heat-killed bacteria. the monoclonal antibodies were characterized by enzyme immunoassay and by western and dot blot analyses and were investigated for their potential use for ...199910225830
induction of heat shock proteins in response to primary alcohols in acinetobacter calcoaceticus.cells of acinetobacter calcoaceticus 69-v, a species able to metabolize a range of aliphatic hydrocarbons and alcohols, were confronted with ethanol, butanol, hexanol or heat shock during growth on acetate as sole source of carbon and energy. the primary alcohols and the heat shock led to the synthesis of new proteins or amplified expression of specific, common and general proteins, which were detected by silver staining after two-dimensional gel electrophoresis. some of the alcohol-inducible pr ...199910344248
structures and properties of gellan polymers produced by sphingomonas paucimobilis atcc 31461 from lactose compared with those produced from glucose and from cheese wheythe dairy industry produces large quantities of whey as a by-product of cheese production and is increasingly looking for new ways to utilize this waste product. gellan gum is reliably produced by sphingomonas paucimobilis in growth media containing lactose, a significant component of cheese whey, as a carbon source. we studied and compared polysaccharide biosynthesis by s. paucimobilis atcc 31461 in media containing glucose, lactose (5 to 30 g/liter), and sweet cheese whey. we found that alteri ...199910347031
characterization of a pseudomonas putida allylic alcohol dehydrogenase induced by growth on 2-methyl-3-buten-2-ol.we have been working to develop an enzymatic assay for the alcohol 2-methyl-3-buten-2-ol (232-mb), which is produced and emitted by certain pines. to this end we have isolated the soil bacterium pseudomonas putida mb-1, which uses 232-mb as a sole carbon source. strain mb-1 contains inducible 3-methyl-2-buten-1-ol (321-mb) and 3-methyl-2-buten-1-al dehydrogenases, suggesting that 232-mb is metabolized by isomerization to 321-mb followed by oxidation. 321-mb dehydrogenase was purified to near-hom ...199910347052
enhancement of solubilization and biodegradation of polyaromatic hydrocarbons by the bioemulsifier alasan.alasan, a high-molecular-weight bioemulsifier complex of an anionic polysaccharide and proteins that is produced by acinetobacter radioresistens ka53 (s. navon-venezia, z. zosim, a. gottlieb, r. legmann, s. carmeli, e. z. ron, and e. rosenberg, appl. environ. microbiol. 61:3240-3244, 1995), enhanced the aqueous solubility and biodegradation rates of polyaromatic hydrocarbons (pahs). in the presence of 500 microg of alasan ml-1, the apparent aqueous solubilities of phenanthrene, fluoranthene, and ...199910347063
genetic analysis of a chromosomal region containing vana and vanb, genes required for conversion of either ferulate or vanillate to protocatechuate in and vanb form an oxygenative demethylase that converts vanillate to protocatechuate in microorganisms. ferulate, an abundant phytochemical, had been shown to be metabolized through a vanillate intermediate in several pseudomonas isolates, and biochemical evidence had indicated that vanillate also is an intermediate in ferulate catabolism by acinetobacter. genetic evidence supporting this conclusion was obtained by characterization of mutant acinetobacter strains blocked in catabolism of bot ...199910348863
substitution, insertion, deletion, suppression, and altered substrate specificity in functional protocatechuate 3,4-dioxygenases.protocatechuate 3,4-dioxygenase is a member of a family of bacterial enzymes that cleave the aromatic rings of their substrates between two adjacent hydroxyl groups, a key reaction in microbial metabolism of varied environmental chemicals. in an appropriate genetic background, it is possible to select for acinetobacter strains containing spontaneous mutations blocking expression of pcah or -g, genes encoding the alpha and beta subunits of protocatechuate 3, 4-dioxygenase. the crystal structure o ...199910515940
rapid identification of burkholderia pseudomallei in blood cultures by a monoclonal antibody assay.burkholderia pseudomallei is the causative agent of melioidosis. in northeast thailand, this gram-negative bacterium is a major cause of mortality from septicemia. the definitive diagnosis of this disease is made by bacterial culture. in this study, we produced a monoclonal antibody (mab) specific to the 30-kda protein of b. pseudomallei by in vivo and in vitro immunization of balb/c mice with a crude culture filtrate antigen. the mab could directly agglutinate with all 243 clinical isolates of ...199910523570
ciprofloxacin in treatment of nosocomial meningitis in neonates and in infants: report of 12 cases and review.twelve cases of neonatal and infant nosocomial meningitis treated with intravenous ciprofloxacin in doses of 10 to 60 mg/kg/day are described. four neonates were 21 to 28 days old and eight infants were 2 to 6 months old. six presented with gram-negative meningitis: escherichia coli (2), salmonella enteritidis (1), acinetobacter calcoaceticus (1), two with two organisms, and (h. influenzae plus staphylococcus epidermidis, acinetobacter spp. plus s. epidermidis), and six were attributable to gram ...199910529884
identification of a transcriptional activator (chnr) and a 6-oxohexanoate dehydrogenase (chne) in the cyclohexanol catabolic pathway in acinetobacter sp. strain ncimb 9871 and localization of the genes that encode them.we identified chnr, a gene encoding an arac-xyls type of transcriptional activator that regulates the expression of chnb, the structural gene for cyclohexanone monooxygenase (chmo) in acinetobacter sp. strain ncimb 9871. the gene sequence of chne, which encodes an nadp(+)-linked 6-oxohexanoate dehydrogenase, the enzyme catalyzing the fifth step of cyclohexanol degradation, was also determined. the gene arrangement is chnb-chne-chnr. the predicted molecular masses of the three polypeptides were v ...199910543838
a novel lipolytic enzyme located in the outer membrane of pseudomonas aeruginosa.a lipase-negative deletion mutant of pseudomonas aeruginosa pao1 still showed extracellular lipolytic activity toward short-chain p-nitrophenylesters. by screening a genomic dna library of p. aeruginosa pao1, an esterase gene, esta, was identified, cloned, and sequenced, revealing an open reading frame of 1,941 bp. the product of esta is a 69.5-kda protein, which is probably processed by removal of an n-terminal signal peptide to yield a 67-kda mature protein. a molecular mass of 66 kda was dete ...199910559163
transcriptional analysis of the rubrerythrin and superoxide dismutase genes of clostridium perfringens.we cloned and sequenced a 2.7-kb fragment of chromosomal dna from clostridium perfringens containing the superoxide dismutase-encoding gene, sod. previously, rubrerythrin from c. perfringens had been isolated and its gene (rbr) had been cloned (y. lehmann, l. meile, and m. teuber, j. bacteriol. 178:7152-7158, 1996). northern blot experiments revealed a length of approximately 800 bases for each transcript of rbr and sod of c. perfringens. thus, rbr and sod each represent a monocistronic operon. ...199910559182
autoantibodies to brain components and antibodies to acinetobacter calcoaceticus are present in bovine spongiform encephalopathy.bovine spongiform encephalopathy (bse) is a neurological disorder, predominantly of british cattle, which belongs to the group of transmissible spongiform encephalopathies together with creutzfeldt-jakob disease (cjd), kuru, and scrapie. autoantibodies to brain neurofilaments have been previously described in patients with cjd and kuru and in sheep affected by scrapie. spongiform-like changes have also been observed in chronic experimental allergic encephalomyelitis, at least in rabbits and guin ...199910569779
characterization of activity and expression of isocitrate lyase in mycobacterium avium and mycobacterium tuberculosis.analysis by two-dimensional gel electrophoresis revealed that mycobacterium avium expresses several proteins unique to an intracellular infection. one abundant protein with an apparent molecular mass of 50 kda was isolated, and the n-terminal sequence was determined. it matches a sequence in the m. tuberculosis database (sanger) with similarity to the enzyme isocitrate lyase of both corynebacterium glutamicum and rhodococcus fascians. only marginal similarity was observed between this open readi ...199910572116
active-site structure of the soluble quinoprotein glucose dehydrogenase complexed with methylhydrazine: a covalent cofactor-inhibitor complex.soluble glucose dehydrogenase (s-gdh) from the bacterium acinetobacter calcoaceticus is a classical quinoprotein. it requires the cofactor pyrroloquinoline quinone (pqq) to catalyze the oxidation of glucose to gluconolactone. the precise catalytic role of pqq in s-gdh and several other pqq-dependent enzymes has remained controversial because of the absence of comprehensive structural data. we have determined the crystal structure of a ternary complex of s-gdh with pqq and methylhydrazine, a comp ...199910518528
bacterial populations in an anthropogenically disturbed stream: comparison of different seasons.abstract to determine the effects of environmental changes on stream bacterial populations, assemblage- and population-level measurements were compared between an anthropogenically disturbed stream and an undisturbed reference stream during different seasons. physical and chemical variables monitored at two disturbed sites from a stream affected by multiple environmental perturbations confirmed discernibly different water quality from three reference sites: two from an adjacent, undisturbed wate ...199910541785
natural competence for dna transformation by legionella pneumophila and its association with expression of type iv pili.we have recently described the expression of two pili of different lengths on the surface of legionella pneumophila (b. j. stone and y. abu kwaik, infect. immun. 66:1768-1775, 1998). production of long pili requires a functional pilel locus, encoding a type iv pilin protein. since type iv pili in neisseria gonorrhoeae are associated with competence for dna transformation, we examined the competence of l. pneumophila for dna transformation under conditions that allowed the expression of type iv p ...199910049368
cloning, expression, and nucleotide sequence of the pseudomonas aeruginosa 142 ohb genes coding for oxygenolytic ortho dehalogenation of halobenzoates.we have cloned and characterized novel oxygenolytic ortho-dehalogenation (ohb) genes from 2-chlorobenzoate (2-cba)- and 2,4-dichlorobenzoate (2,4-dcba)-degrading pseudomonas aeruginosa 142. among 3,700 escherichia coli recombinants, two clones, dh5alphaf'(pod22) and dh5alphaf'(pod33), converted 2-cba to catechol and 2,4-dcba and 2,5-dcba to 4-chlorocatechol. a subclone of pod33, plasmid pe43, containing the 3,687-bp minimized ohb dna region conferred to p. putida pb2440 the ability to grow on 2- ...199910224014
adenosylcobalamin-mediated methyl transfer by toluate cis-dihydrodiol dehydrogenase of the tol plasmid pww0.we identified and characterized a methyl transfer activity of the toluate cis-dihydrodiol (4-methyl-3,5-cyclohexadiene-cis-1, 2-diol-1-carboxylic acid) dehydrogenase of the tol plasmid pww0 towards toluene cis-dihydrodiol (3-methyl-4,5-cyclohexadiene-cis-1, 2-diol). when the purified enzyme from the recombinant escherichia coli containing the xyll gene was incubated with toluene cis-dihydrodiol in the presence of nad+, the end products differed depending on the presence of adenosylcobalamin (coe ...199910217792
in vitro and in vivo antimicrobial activities of t-3811me, a novel des-f(6)-quinolone.the in vitro and in vivo activities of t-3811me, a novel des-f(6)-quinolone, were evaluated in comparison with those of some fluoroquinolones, including a newly developed one, trovafloxacin. t-3811, a free base of t-3811me, showed a wide range of antimicrobial spectra, including activities against chlamydia trachomatis, mycoplasma pneumoniae, and mycobacterium tuberculosis. in particular, t-3811 exhibited potent activity against various gram-positive cocci, with mics at which 90% of the isolates ...199910223917
integron- and carbenicillinase-mediated reduced susceptibility to amoxicillin-clavulanic acid in isolates of multidrug-resistant salmonella enterica serotype typhimurium dt104 from french patients.fifty-seven salmonella enterica serotype typhimurium (s. typhimurium) isolates were collected from human patients in two french hospitals, hôpital antoine béclère (clamart, france) and hôpital bicêtre (le kremlin-bicêtre, france), between 1996 and 1997. thirty of them (52 percent) were resistant to amino-, carbeni-, and ureidopenicillins, had reduced susceptibility to amoxicillin-clavulanic acid, were susceptible to cephalothin, and were resistant to sulfonamides, streptomycin, chloramphenicol, ...199910223920
in vivo efficacies of combinations of beta-lactams, beta-lactamase inhibitors, and rifampin against acinetobacter baumannii in a mouse pneumonia model.the effects of various regimens containing combinations of beta-lactams, beta-lactam inhibitor(s), and rifampin were assessed in a recently described mouse model of acinetobacter baumannii pneumonia (m. l. joly-guillou, m. wolff, j. j. pocidalo, f. walker, and c. carbon, antimicrob. agents chemother. 41:345-351, 1997). two aspects of the therapeutic response were studied: the kinetics of the bactericidal effect (treatment was initiated 3 h after intratracheal inoculation, and bacterial counts we ...199910348761
phylogenetic structures of the genus acinetobacter based on gyrb sequences: comparison with the grouping by dna-dna hybridization.the phylogenetic relationships of 49 acinetobacter strains, 46 of which have previously been classified into 18 genomic species by dna-dna hybridization studies, were investigated using the nucleotide sequence of gyrb, the structural gene for the dna gyrase b subunit. the phylogenetic tree showed linkages between genomic species 1 (acinetobacter calcoaceticus), 2 (acinetobacter baumannii), 3 and tu13; genomic species 6, bj15, bj16 and bj17; genomic species 5, bj13 (synonym of tu14) and bj14; gen ...199910028249
adhesion of acinetobacter venetianus to diesel fuel droplets studied with in situ electrochemical and molecular probesthe adhesion of a recently described species, acinetobacter venetianus ve-c3 (f. di cello, m. pepi, f. baldi, and r. fani, res. microbiol. 148:237-249, 1997), to diesel fuel (a mixture of c12 to c28 n-alkanes) and n-hexadecane was studied and compared to that of acinetobacter sp. strain rag-1, which is known to excrete the emulsifying lipopolysaccharide, emulsan. oxygen consumption rates, biomass, cell hydrophobicity, electrophoretic mobility, and zeta potential were measured for the two strains ...199910223998
evidence for a chemiosmotic model of dehalorespiration in desulfomonile tiedjei dcb-1.desulfomonile tiedjei dcb-1, a sulfate-reducing bacterium, conserves energy for growth from reductive dehalogenation of 3-chlorobenzoate by an uncharacterized chemiosmotic process. respiratory electron transport components were examined in d. tiedjei cells grown under conditions for reductive dehalogenation, pyruvate fermentation, and sulfate reduction. reductive dehalogenation was inhibited by the respiratory quinone inhibitor 2-heptyl-4-hydroxyquinoline n-oxide, suggesting that a respiratory q ...19999864310
genetic and biochemical characterization of a 2-pyrone-4, 6-dicarboxylic acid hydrolase involved in the protocatechuate 4, 5-cleavage pathway of sphingomonas paucimobilis syk-6.sphingomonas paucimobilis syk-6 is able to grow on a wide variety of dimeric lignin compounds with guaiacyl moieties, which are converted into protocatechuate by the actions of lignin degradation enzymes in this strain. protocatechuate is a key metabolite in the syk-6 degradation of lignin compounds with guaiacyl moieties, and it is thought that it degrades to pyruvate and oxaloacetate via the protocatechuate 4,5-cleavage pathway. in a 10.5-kb ecori fragment carrying the protocatechuate 4,5-diox ...19999864312
characterization of biosynthetic enzymes for ectoine as a compatible solute in a moderately halophilic eubacterium, halomonas elongata.1,4,5,6-tetrahydro-2-methyl-4-pyrimidinecarboxylic acid (ectoine) is an excellent osmoprotectant. the biosynthetic pathway of ectoine from aspartic beta-semialdehyde (asa), in halomonas elongata, was elucidated by purification and characterization of each enzyme involved. 2,4-diaminobutyrate (daba) aminotransferase catalyzed reversively the first step of the pathway, conversion of asa to daba by transamination with l-glutamate. this enzyme required pyridoxal 5'-phosphate and potassium ions for i ...19999864317
fate of free dna and transformation of the oral bacterium streptococcus gordonii dl1 by plasmid dna in human saliva.competitive pcr was used to monitor the survival of a 520-bp dna target sequence from a recombinant plasmid, pvacmc1, after admixture of the plasmid with freshly sampled human saliva. the fraction of the target remaining amplifiable ranged from 40 to 65% after 10 min of exposure to saliva samples from five subjects and from 6 to 25% after 60 min of exposure. pvacmc1 plasmid dna that had been exposed to degradation by fresh saliva was capable of transforming naturally competent streptococcus gord ...19999872752
in situ, real-time catabolic gene expression: extraction and characterization of naphthalene dioxygenase mrna transcripts from groundwater.we developed procedures for isolating and characterizing in situ-transcribed mrna from groundwater microorganisms catabolizing naphthalene at a coal tar waste-contaminated site. groundwater was pumped through 0.22-microm-pore-size filters, which were then frozen in dry ice-ethanol. rna was extracted from the frozen filters by boiling sodium dodecyl sulfate lysis and acidic phenol-chloroform extraction. transcript characterization was performed with a series of pcr primers designed to amplify nah ...19999872763
plasmid-encoded anthranilate synthase (trpeg) in buchnera aphidicola from aphids of the family pemphigidae.buchnera aphidicola is an obligate intracellular symbiont of aphids. one of its proposed functions is the synthesis of essential amino acids, nutrients required by aphids but deficient in their diet of plant phloem sap. the genetic organization of the tryptophan pathway in buchnera from proliferous aphids of the family aphididae has previously been shown to reflect a capacity to overproduce this essential amino acid (c.-y. lai, l. baumann, and p. baumann, proc. natl. acad. sci. usa 91:3819-3823, ...19999872768
bacterial adhesion to soil contaminants in the presence of surfactantsit has been proposed that addition of surfactants to contaminated soil enhances the solubility of target compounds; however, surfactants may simultaneously reduce the adhesion of bacteria to hydrophobic surfaces. if the latter mechanism is important for the biodegradation of virtually insoluble contaminants, then the use of surfactants may not be beneficial. the adhesion of a mycobacterium strain and a pseudomonas strain, isolated from a creosote-contaminated soil, to the surfaces of highly visc ...19999872775
the alpha subunit of toluene dioxygenase from pseudomonas putida f1 can accept electrons from reduced ferredoxintol but is catalytically inactive in the absence of the beta subunit.the oxygenase component of toluene dioxygenase from pseudomonas putida f1 is an iron-sulfur protein (isptol) consisting of alpha (todc1) and beta (todc2) subunits. purified todc1 gave absorbance and electron paramagnetic resonance spectra identical to those given by purified isptol. todc1 was reduced by nadh and catalytic amounts of reductasetol and ferredoxintol. reduced todc1 did not oxidize toluene, and catalysis was strictly dependent on the presence of purified todc2.19999872799
production of wax esters during aerobic growth of marine bacteria on isoprenoid compoundsthis paper describes the production of isoprenoid wax esters during the aerobic degradation of 6,10,14-trimethylpentadecan-2-one and phytol by four bacteria (acinetobacter sp. strain phy9, pseudomonas nautica [ip85/617], marinobacter sp. strain cab [dsmz 11874], and marinobacter hydrocarbonoclasticus [atcc 49840]) isolated from the marine environment. different pathways are proposed to explain the formation of these compounds. in the case of 6,10, 14-trimethylpentadecan-2-one, these esters resul ...19999872783
characterization and nucleotide sequence of carb-6, a new carbenicillin-hydrolyzing beta-lactamase from vibrio cholerae.a clinical strain of vibrio cholerae non-o1 non-o139 isolated in france produced a new beta-lactamase with a pi of 5.35. the purified enzyme, with a molecular mass of 33,000 da, was characterized. its kinetic constants show it to be a carbenicillin-hydrolyzing enzyme comparable to the five previously reported carb beta-lactamases and to sar-1, another carbenicillin-hydrolyzing beta-lactamase that has a pi of 4.9 and that is produced by a v. cholerae strain from tanzania. this beta-lactamase is d ...19999925522
vig-1, a new fish gene induced by the rhabdovirus glycoprotein, has a virus-induced homologue in humans and shares conserved motifs with the moaa family.we used mrna differential display methodology to analyze the shift of transcription profile induced by the fish rhabdovirus, viral hemorrhagic septicemia virus (vhsv), in rainbow trout leukocytes. we identified and characterized a new gene which is directly induced by vhsv. this vhsv-induced gene (vig-1) encodes a 348-amino-acid protein. vig-1 is highly expressed during the experimental disease in lymphoid organs of the infected fish. intramuscular injection of a plasmid vector expressing the vi ...19999971762
spread of amikacin resistance in acinetobacter baumannii strains isolated in spain due to an epidemic strain.sixteen amikacin-resistant clinical acinetobacter baumannii isolates from nine different hospitals in spain were investigated to determine whether the high incidence of amikacin-resistant a. baumannii was due to the dissemination of an amikacin-resistant strain or to the spread of an amikacin resistance gene. the epidemiological relationship studied by repetitive extragenic palindromic pcr and low-frequency restriction analysis of chromosomal dna showed that the same clone was isolated in eight ...19999986846
dust-borne bacteria in animal sheds, schools and children's day care centres.a total of 316 bacterial strains, including psychrophiles, mesophiles and thermophiles, were isolated and identified from indoor dusts in schools, children's day care centres and animal sheds. several species which had not previously been reported from indoor environments were found: sphingomonas, brevibacterium, nocardiopsis, deinococcus and rhodococcus/gordona. a new psychrophilic actinomycete genus was also found in animal sheds, representing a new undescribed peptidoglycan type and an unusua ...199910212408
random mutagenesis by recombinational capture of pcr products in bacillus subtilis and acinetobacter calcoaceticus.we describe a general method for random mutagenesis of cloned genes by error-prone pcr or dna shuffling that eliminates the need for post-amplification subcloning following each cycle of mutagenesis. this method exploits the highly efficient and recombinogenic nature of dna uptake during natural transformation in the gram-positive bacterium bacillus subtilis and the gram-negative bacterium acinetobacter calcoaceticus. plasmid systems were designed that allow capture of pcr-amplified dna fragment ...19999927739
cloning and sequence analysis of the lipase and lipase chaperone-encoding genes from acinetobacter calcoaceticus rag-1, and redefinition of a proteobacterial lipase family and an analogous lipase chaperone family.the genes encoding the lipase (lipa) and lipase chaperone (lipb) from acinetobacter calcoaceticus rag-1 were cloned and sequenced. the genes were isolated from a genomic dna library by complementation of a lipase-deficient transposon mutant of the same strain. transposon insertion in this mutant and three others was mapped to a single site in the chaperone gene. the deduced amino acid (aa) sequences for the lipase and its chaperone were found to encode mature proteins of 313 aa (32.5kda) and 347 ...199910216267
in vitro activities of aminomethyl-substituted analogs of novel tetrahydrofuranyl 188,624, cl 190,294, and cl 191,121 are novel aminomethyl tetrahydrofuranyl (thf)-1 beta-methylcarbapenems. the in vitro antibacterial activities of these thf carbapenems were evaluated and compared with those of biapenem, imipenem, and meropenem against 554 recent clinical isolates obtained from geographically distinct medical centers across north america. the antibacterial activities of the thf carbapenems were equivalent to that of biapenem, and the thf carbapenems were slightly more activ ...199910049250
complete sequence of a 184-kilobase catabolic plasmid from sphingomonas aromaticivorans f199.the complete 184,457-bp sequence of the aromatic catabolic plasmid, pnl1, from sphingomonas aromaticivorans f199 has been determined. a total of 186 open reading frames (orfs) are predicted to encode proteins, of which 79 are likely directly associated with catabolism or transport of aromatic compounds. genes that encode enzymes associated with the degradation of biphenyl, naphthalene, m-xylene, and p-cresol are predicted to be distributed among 15 gene clusters. the unusual coclustering of gene ...199910049392
cloning and characterization of polyphosphate kinase and exopolyphosphatase genes from pseudomonas aeruginosa 8830.pseudomonas aeruginosa accumulates polyphosphates in response to nutrient limitations. to elucidate the function of polyphosphate in this microorganism, we have investigated polyphosphate metabolism by isolating from p. aeruginosa 8830 the genes encoding polyphosphate kinase (ppk) and exopolyphosphatase (ppx), which are involved in polyphosphate synthesis and degradation, respectively. the 690- and 506-amino-acid polypeptides encoded by the two genes have been expressed in escherichia coli and p ...199910224002
molecular characterization of the genes pcag and pcah, encoding protocatechuate 3,4-dioxygenase, which are essential for vanillin catabolism in pseudomonas sp. strain hr199.pseudomonas sp. strain hr199 is able to utilize eugenol (4-allyl-2-methoxyphenol), vanillin (4-hydroxy-3-methoxybenzaldehyde), or protocatechuate as the sole carbon source for growth. mutants of this strain which were impaired in the catabolism of vanillin but retained the ability to utilize eugenol or protocatechuate were obtained after nitrosoguanidine mutagenesis. one mutant (sk6169) was used as recipient of a pseudomonas sp. strain hr199 genomic library in cosmid pvk100, and phenotypic compl ...199910049847
engineering a chimeric pyrroloquinoline quinone glucose dehydrogenase: improvement of edta tolerance, thermal stability and substrate engineered escherichia coli pqq glucose dehydrogenase (pqqgdh) with improved enzymatic characteristics was constructed by substituting and combining the gene-encoding protein regions responsible for edta tolerance, thermal stability and substrate specificity. the protein region responsible for complete edta tolerance in acinetobacter calcoaceticus, which is recognized as the indicator of high stability in co-factor binding, was elucidated. the region is located between 32 and 59% from the n-t ...199910065712
molecular analysis of a novel methanesulfonic acid monooxygenase from the methylotroph methylosulfonomonas methylovora.methylosulfonomonas methylovora m2 is an unusual gram-negative methylotrophic bacterium that can grow on methanesulfonic acid (msa) as the sole source of carbon and energy. oxidation of msa by this bacterium is carried out by a multicomponent msa monooxygenase (msamo). cloning and sequencing of a 7.5-kbp sphi fragment of chromosomal dna revealed four tightly linked genes encoding this novel monooxygenase. analysis of the deduced msamo polypeptide sequences indicated that the enzyme contains a tw ...199910094704
characterization of mdcr, a regulatory gene of the malonate catabolic system in klebsiella pneumoniae.the klebsiella pneumoniae mdcr gene, which encodes a lysr-type regulator, was overexpressed in escherichia coli. purified mdcr was found to bind specifically to the control region of either the malonate decarboxylase (mdc) genes or mdcr. we have also demonstrated that mdcr is an activator of the expression of the mdc genes, whereas it represses the transcription of the putative control region of mdcr, pmdcr, indicating a negative autoregulatory control.199910094715
use of a murine o-antigen-specific monoclonal antibody to identify acinetobacter strains of unnamed genomic species 13 sensu tjernberg and ursing.a monoclonal antibody against the o-antigenic polysaccharide chain of the lipopolysaccharide (lps) of acinetobacter strains belonging to the unnamed genomic species 13 sensu tjernberg and ursing (13tu) was obtained after immunization of balb/c mice with heat-killed bacteria and was characterized by enzyme immunoassay and western blot analysis, by use of lps and proteinase k-treated bacterial lysates, analyses in which the antibody was shown to be highly specific for the homologous antigen. in ad ...199910325309
emergence of fosfomycin-resistant isolates of shiga-like toxin-producing escherichia coli o26.we evaluated the susceptibilities of 129 shiga-like toxin-producing escherichia coli (stec) isolates to various antibiotics. the numbers of isolates for which mics were high (> or = 128 micrograms/ml) were as follows: 5 for fosfomycin, 14 for ampicillin, 1 for cefaclor, 6 for kanamycin, 22 for tetracycline, and 2 for doxycycline. for two isolates of stec o26 mics of fosfomycin were high (1,024 and 512 micrograms/ml, respectively). conjugation experiments and glutathione s-transferase assays sugg ...199910103182
clavulanate induces expression of the pseudomonas aeruginosa ampc cephalosporinase at physiologically relevant concentrations and antagonizes the antibacterial activity of ticarcillin.although previous studies have indicated that clavulanate may induce ampc expression in isolates of pseudomonas aeruginosa, the impact of this inducer activity on the antibacterial activity of ticarcillin at clinically relevant concentrations has not been investigated. therefore, a study was designed to determine if the inducer activity of clavulanate was associated with in vitro antagonism of ticarcillin at pharmacokinetically relevant concentrations. by the disk approximation methodology, clav ...199910103195
the alkene monooxygenase from xanthobacter strain py2 is closely related to aromatic monooxygenases and catalyzes aromatic monohydroxylation of benzene, toluene, and phenol.the genes encoding the six polypeptide components of the alkene monooxygenase from xanthobacter strain py2 (xamo) have been located on a 4.9-kb fragment of chromosomal dna previously cloned in cosmid pny2. sequencing and analysis of the predicted amino acid sequences indicate that the components of xamo are homologous to those of the aromatic monooxygenases, toluene 2-, 3-, and 4-monooxygenase and benzene monooxygenase, and that the gene order is identical. the genes and predicted polypeptides a ...199910103255
effects of surfactant mixtures, including corexit 9527, on bacterial oxidation of acetate and alkanes in crude oil.mixtures of nonionic and anionic surfactants, including corexit 9527, were tested to determine their effects on bacterial oxidation of acetate and alkanes in crude oil by cells pregrown on these substrates. corexit 9527 inhibited oxidation of the alkanes in crude oil by acinetobacter calcoaceticus atcc 31012, while span 80, a corexit 9527 constituent, markedly increased the oil oxidation rate. another corexit 9527 constituent, the negatively charged dioctyl sulfosuccinate (aot), strongly reduced ...199910103264
phenotypic expression of pcr-generated random mutations in a pseudomonas putida gene after its introduction into an acinetobacter chromosome by natural transformation.localized sets of random point mutations generated by pcr amplification can be transferred efficiently to the chromosome of acinetobacter adp1 (also known as strain bd413) by natural transformation. the technique does not require cloning of pcr fragments in plasmids: pcr-amplified dna fragments are internalized by cells and directly incorporated into their genomes by homologous recombination. previously such procedures for random mutagenesis could be applied only to acinetobacter genes affording ...199910103267
a novel aromatic-ring-hydroxylating dioxygenase from the diterpenoid-degrading bacterium pseudomonas abietaniphila bkme-9.pseudomonas abietaniphila bkme-9 is able to degrade dehydroabietic acid (dha) via ring hydroxylation by a novel dioxygenase. the dita1, dita2, and dita3 genes, which encode the alpha and beta subunits of the oxygenase and the ferredoxin of the diterpenoid dioxygenase, respectively, were isolated and sequenced. the ferredoxin gene is 9. 2 kb upstream of the oxygenase genes and 872 bp upstream of a putative meta ring cleavage dioxygenase gene, ditc. a tn5 insertion in the alpha subunit gene, dita1 ...199910217753
acs1 of haemophilus influenzae type a capsulation locus region ii encodes a bifunctional ribulose 5-phosphate reductase- cdp-ribitol pyrophosphorylase.the serotype-specific, 5.9-kb region ii of the haemophilus influenzae type a capsulation locus was sequenced and found to contain four open reading frames termed acs1 to acs4. acs1 was 96% identical to h. influenzae type b orf1, previously shown to have cdp-ribitol pyrophosphorylase activity (j. van eldere, l. brophy, b. loynds, p. celis, i. hancock, s. carman, j. s. kroll, and e. r. moxon, mol. microbiol. 15:107-118, 1995). low but significant homology to other pyrophosphorylases was only detec ...199910094675
cloning and sequencing of a novel meta-cleavage dioxygenase gene whose product is involved in degradation of gamma-hexachlorocyclohexane in sphingomonas paucimobilis.sphingomonas (formerly pseudomonas) paucimobilis ut26 utilizes gamma-hexachlorocyclohexane (gamma-hch), a halogenated organic insecticide, as a sole source of carbon and energy. in a previous study, we showed that gamma-hch is degraded to chlorohydroquinone (chq) and then to hydroquinone (hq), although the rate of reaction from chq to hq was slow (k. miyauchi, s. k. suh, y. nagata, and m. takagi, j. bacteriol. 180:1354-1359, 1998). in this study, we cloned and characterized a gene, designated li ...199910542173
stereochemical course of biotin-independent malonate decarboxylase catalysis.malonate decarboxylases, which catalyze the conversion of malonate to acetate, can be classified into biotin-dependent and biotin-independent enzymes. in order to reveal the stereochemical course of the reactions catalyzed by the biotin-independent enzymes from acinetobacter calcoaceticus and pseudomonas fluorescens, a chiral substrate, malonate carrying (13)c in one carboxyl group and (3)h at one of the methylene positions, was prepared and used in the reactions catalyzed by these two enzymes. ...199910496981
characteristics of acinetobacter strains (phenotype classification, antibiotic susceptibility and production of beta-lactamases) isolated from haemocultures from patients at the teaching hospital in olomouc.a total of 85 strains of the genus acinetobacter were isolated from haemocultures at the institute of microbiology of the teaching hospital in olomouc over the period january 1993 to june 1997. sixty-two (73.0%) strains of the acinetobacter calcoaceticus-baumannii complex (acb complex) were the most frequent. in 3 (3.5%) strains it was impossible to decide whether they belonged to the acb complex. other acinetobacter species were represented by 20 (23.5%) strains. the greatest amount (28.2%) of ...199910743729
functional evaluation of the genes involved in malonate decarboxylation by acinetobacter calcoaceticus.the genomic locus containing the potential repressor gene mdcy (inactivated by a putative is3 element) and the mdclmacdegbh genes from acinetobacter calcoaceticus was cloned and sequenced. in order to evaluate the biochemical function of the protein components, the genes were expressed independently and their activities predicted by database analysis. the mdca gene product, the alpha subunit, was found to be malonate/acetyl-coa transferase and the mdcd gene product, the beta subunit, was found t ...199910561613
the discriminatory power of ribo-pcr compared to conventional ribotyping for epidemiological purposes.molecular typing techniques have become increasingly important for confirmation of epidemiological relationships and delimitation of nosocomial outbreaks. the discriminatory power of the two dna-based typing methods, conventional ribotyping and ribo-pcr, was assessed to distinguish between selected strains of acinetobacter calcoaceticus, enterobacter cloacae, serratia marcescens and pseudomonas aeruginosa. overall, conventional ribotyping was more discriminatory than ribo-pcr.199910660137
isolation and partial characterization of bac201: a plasmid-associated bacteriocin-like inhibitory substance from staphylococcus aureus ab201.staphylococcus aureus ab201, a clinical isolate from wound pus, produced a bacteriocin-like inhibitory substance termed as bac201 that exhibited a broad-spectrum activity against both gram-positive as well as gram-negative bacteria. among gram-negative bacteria it was active against neisseria meningitidis and acinetobacter calcoaceticus both being gram-negative cocci. inhibition due to the effect of organic acids, hydrogen peroxide, or bacteriophages was excluded. this inhibitory substance could ...199910629973
[the clinical picture, treatment and prognosis of meningitis due to anaerobic and nonfermentative bacteria].the study of incidence, clinical manifestations and prognosis of meningitis with anaerobic and non-fermentative bacteria.199910756944
the phn genes of burkholderia sp. strain rp007 constitute a divergent gene cluster for polycyclic aromatic hydrocarbon catabolism.cloning and molecular ecological studies have underestimated the diversity of polycyclic aromatic hydrocarbon (pah) catabolic genes by emphasizing classical nah-like (nah, ndo, pah, and dox) sequences. here we report the description of a divergent set of pah catabolic genes, the phn genes, which although isofunctional to the classical nah-like genes, show very low homology. this phn locus, which contains nine open reading frames (orfs), was isolated on an 11.5-kb hindiii fragment from phenanthre ...19999882667
a sensitive, viable-colony staining method using nile red for direct screening of bacteria that accumulate polyhydroxyalkanoic acids and other lipid storage compounds.the oxazine dye nile blue a and its fluorescent oxazone form, nile red, were used to develop a simple and highly sensitive staining method to detect poly(3-hydroxybutyric acid) and other polyhydroxyalkanoic acids (phas) directly in growing bacterial colonies. in contrast to previously described methods, these dyes were directly included in the medium at concentrations of only 0.5 microgram/ml, and growth of the cells occurred in the presence of the dyes. this allowed an estimation of the presenc ...19999914303
poly-3-hydroxybutyrate degradation in rhizobium (sinorhizobium) meliloti: isolation and characterization of a gene encoding 3-hydroxybutyrate dehydrogenase.we have cloned and sequenced the 3-hydroxybutyrate dehydrogenase-encoding gene (bdha) from rhizobium (sinorhizobium) meliloti. the gene has an open reading frame of 777 bp that encodes a polypeptide of 258 amino acid residues (molecular weight 27,177, pi 6.07). the r. meliloti bdh protein exhibits features common to members of the short-chain alcohol dehydrogenase superfamily. bdha is the first gene transcribed in an operon that also includes xdha, encoding xanthine oxidase/dehydrogenase. transc ...19999922248
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