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genetic organization of the mau gene cluster in methylobacterium extorquens am1: complete nucleotide sequence and generation and characteristics of mau mutants.the nucleotide sequence of the methylamine utilization (mau) gene region from methylobacterium extorquens am1 was determined. open reading frames for 11 genes (maufbedacjglmn) were found, all transcribed in the same orientation. the maub, maua, and mauc genes encode the periplasmic methylamine dehydrogenase (madh) large and small subunit polypeptides and amicyanin, respectively. the products of maud, maug, maul, and maum were also predicted to be periplasmic. the products of mauf, maue, and maun ...19948021187
nucleotide sequence and functional analysis of the meta-cleavage pathway involved in biphenyl and polychlorinated biphenyl degradation in pseudomonas sp. strain kks102.pseudomonas sp. strain kks102 is able to degrade biphenyl and polychlorinated biphenyls via the meta-cleavage pathway. we sequenced the upstream region of the bpha1a2a3bcd (open reading frame 1 [orf1]) a4 and found four orfs in this region. as the deduced amino acid sequences of the first, second, and third orfs are homologous to the meta-cleavage enzymes from pseudomonas sp. strain cf600 (v. shingler, j. powlowski, and u. marklund, j. bacteriol. 174:711-724, 1992), these orfs have been named bp ...19948021212
inability of muconate cycloisomerases to cause dehalogenation during conversion of 2-chloro-cis,cis-muconate.the conversion of 2-chloro-cis,cis-muconate by muconate cycloisomerase from pseudomonas putida prs2000 yielded two products which by nuclear magnetic resonance spectroscopy were identified as 2-chloro- and 5-chloromuconolactone. high-pressure liquid chromatography analyses showed the same compounds to be formed also by muconate cycloisomerases from acinetobacter calcoaceticus adp1 and pseudomonas sp. strain b13. during 2-chloro-cis,cis-muconate turnover by the enzyme from p. putida, 2-chloromuco ...19948021223
cross-regulation by xylr and dmpr activators of pseudomonas putida suggests that transcriptional control of biodegradative operons evolves independently of catabolic genes.the pu promoter of the toluene degradation plasmid pww0 of pseudomonas putida drives expression of an operon involved in the sequential oxidation of toluene and m- and p-xylenes to benzoate and toluates, respectively. similarly, the po promoter of plasmid pvi150 controls expression of an operon of pseudomonas sp. strain cf600 which is required for the complete catabolism of phenol and cresols. these promoters, which both belong to the sigma 54-dependent class, are regulated by their cognate acti ...19948051017
degradation of alkylphenol ethoxylates by pseudomonas sp. strain tr01.an alkylphenol ethoxylate-degrading bacterium was isolated from activated sludge of a municipal sewage treatment plant by enrichment culture. this organism was found to belong to the genus pseudomonas; since no corresponding species was identified, we designated it as pseudomonas sp. strain tr01. this strain had an optimal temperature and ph of 30 degrees c and 7, respectively, for both growth and the degradation of triton n-101 (a nonylphenol ethoxylate in which the average number of ethylene o ...19948074508
purification and characterization of thermostable and nonthermostable 2-haloacid dehalogenases with different stereospecificities from pseudomonas sp. strain yl.two novel hydrolytic dehalogenases, thermostable l-2-haloacid dehalogenase (l-dex) inducibly synthesized by 2-chloropropionate (2-cpa) and nonthermostable dl-2-haloacid dehalogenase (dl-dex) induced by 2-chloroacrylate, were purified to homogeneity from pseudomonas sp. strain yl. dl-dex consisted of a monomer with a molecular weight of about 36,000 and catalyzed the dehalogenation of l and d isomers of 2-cpa to produce d- and l-lactates, respectively. it acted on 2-haloalkanoic acids with a carb ...19948074519
evidence for a novel pathway in the degradation of fluorene by pseudomonas sp. strain f274.a fluorene-utilizing microorganism, identified as a species of pseudomonas, was isolated from soil severely contaminated from creosote use and was shown to accumulate six major metabolites from fluorene in washed-cell incubations. five of these products were identified as 9-fluorenol, 9-fluorenone, (+)-1,1a-dihydroxy-1-hydro-9-fluorenone, 8-hydroxy-3,4-benzocoumarin, and phthalic acid. this last compound was also identified in growing cultures supported by fluorene. fluorene assimilation into ce ...19948074523
azoreductase activity in bacteria associated with the greening of instant chocolate puddings.pseudomonas sp. strain azr1 and klebsiella sp. strain azr2 were isolated from reconstituted instant chocolate puddings that had turned green and were found to have azoreductase activity. this activity was inducible and nadh dependent. differences in dye reduction rates between the two strains were apparent, and substrate specificity related to dye structure was observed.19948085839
insertion sequence is6100 on plasmid poad2, which degrades nylon oligomers.the nucleotide sequence of repeated sequence i, which appears in five regions on nylon oligomer-degrading plasmid poad2, harbored in flavobacterium sp. strain k172, was determined. the five regions of repeated sequence i had 880 bp of identical sequence, and the sequence was identical to that of is6100, an insertion sequence classified in the is6 family, initially found in mycobacterium fortuitum. sequences homologous to that of is6100 were found for another nylon oligomer-degrading plasmid, pna ...19948106333
analysis of the binding site of the lysr-type transcriptional activator tcbr on the tcbr and tcbc divergent promoter sequences.the tcbr transcriptional activator protein, which is encoded by the tcbr gene of pseudomonas sp. strain p51 (j. r. van der meer, a. c. j. frijters, j. h. j. leveau, r. i. l. eggen, a. j. b. zehnder, and w. m. de vos, j. bacteriol. 173:3700-3708, 1991), was purified from overproducing escherichia coli cells by using a two-step chromatographic procedure. subsequent use of tcbr in gel mobility shift assays with progressively shortened portions of a dna fragment containing the divergent promoter seq ...19948144450
sensing of aromatic compounds by the dmpr transcriptional activator of phenol-catabolizing pseudomonas sp. strain cf600.the dmp operon of the pvi150 catabolic plasmid of pseudomonas sp. strain cf600 encodes the enzymes involved in the catabolism of phenol and methylphenols. the regulator of this dmp pathway, dmpr, is a member of the ntrc family of transcriptional activators and controls transcription of the dmp operon in response to aromatic effector compounds (v. shingler, m. bartilson, and t. moore, j. bacteriol. 175:1596-1604, 1993). using a lux gene fusion reporter system, in which the dmpr-regulated operon p ...19948132448
identification of the bpha4 gene encoding ferredoxin reductase involved in biphenyl and polychlorinated biphenyl degradation in pseudomonas sp. strain kks102.the nucleotide sequence of the downstream region of the bph operon from pseudomonas sp. strain kks102 was determined. two open reading frames (orf1 and orf2) were found in this region, and the deduced amino acid sequence of orf2 showed homology with the sequences of four ferredoxin reductases of dioxygenase systems. when this region was inserted just upstream of the bph operon, which does not contain a gene encoding ferredoxin reductase, biphenyl dioxygenase activity was detected. the 24- and 44 ...19948132464
metabolism of both 4-chlorobenzoate and toluene under denitrifying conditions by a constructed bacterial strain.t1, a dentrifying bacterium originally isolated for its ability to grow on toluene, can also metabolize 4-hydroxybenzoate and other aromatic compounds under denitrifying conditions. a cosmid clone carrying the three genes that code for the 4-chlorobenzoate dehalogenase enzyme complex isolated from the aerobic bacterium pseudomonas sp. strain cbs3 was successfully conjugated into strain t1. the cloned enzyme complex catalyzes the hydrolytic dechlorination of 4-chlorobenzoate to 4-hydroxybenzoate. ...19948161190
identification of a major soluble protein in mitochondria from nonphotosynthetic tissues as nad-dependent formate dehydrogenase.in many plant species, one of the most abundant soluble proteins (as judged by two-dimensional polyacrylamide gel electrophoresis) in mitochondria from nongreen tissues is a 40-kd polypeptide that is relatively scarce in mitochondria from photosynthetic tissues. cdna sequences encoding this polypeptide were isolated from a lambda gt11 cdna expression library from potato (solanum tuberosum l.) by screening with a specific antibody raised against the 40-kd polypeptide. the cdna sequence contains a ...19938278546
enhanced biodegradation of polychlorinated biphenyls after site-directed mutagenesis of a biphenyl dioxygenase gene.biphenyl dioxygenase catalyzes the first step in the aerobic degradation of polychlorinated biphenyls (pcbs). the nucleotide and amino acid sequences of the biphenyl dioxygenases from two pcb-degrading strains (pseudomonas sp. strain lb400 and pseudomonas pseudoalcaligenes kf707) were compared. the sequences were found to be nearly identical, yet these enzymes exhibited dramatically different substrate specificities for pcbs. site-directed mutagenesis of the lb400 bpha gene resulted in an enzyme ...19938285689
detection of polychlorinated biphenyl degradation genes in polluted sediments by direct dna extraction and polymerase chain reaction.it was the aim of this study to specifically detect the dna sequences for the bphc gene, the meta-cleavage enzyme of the aerobic catabolic pathway for biphenyl and polychlorinated biphenyl degradation, in aquatic sediments without prior cultivation of microorganisms by using extraction of total dna, pcr amplification of bphc sequences, and detection with specific gene probes. the direct dna extraction protocol used was modified to enhance lysis efficiency. crude extracts of dna were further puri ...19938285706
identification and characterization of the treponema pallidum tpn50 gene, an ompa homolog.treponema pallidum is a pathogenic spirochete that has no known genetic exchange mechanisms. in order to identify treponemal genes encoding surface and secreted proteins, we carried out tnphoa mutagenesis of a t. pallidum genomic dna library in escherichia coli. several of the resulting clones expressed enzymatically active t. pallidum-alkaline phosphatase fusion proteins. the dna sequence of the 5' portion of a number of the treponemal genes was obtained and analyzed. a recombinant clone harbor ...19948112835
biodegradation of 4-methyl-5-nitrocatechol by pseudomonas sp. strain dnt.pseudomonas sp. strain dnt degrades 2,4-dinitrotoluene (dnt) by a dioxygenase attack at the 4,5 position with concomitant removal of the nitro group to yield 4-methyl-5-nitrocatechol (mnc). here we describe the mechanism of removal of the nitro group from mnc and subsequent reactions leading to ring fission. washed suspensions of dnt-grown cells oxidized mnc and 2,4,5-trihydroxytoluene (tht). extracts prepared from dnt-induced cells catalyzed the disappearance of mnc in the presence of oxygen an ...19948195105
influence of organic nutrients and cocultures on the competitive behavior of 1,2-dichloroethane-degrading bacteria.the effects of organic nutrients and cocultures on substrate removal by and competitive behavior of 1,2-dichloroethane-degrading bacteria were investigated. xanthobacter autotrophicus gj10 needed biotin for optimal growth on 1,2-dichloroethane. in continuous culture, dilution of biotin to a concentration below 0.2 nm resulted in washout. growth could be restored by inoculation with the 2-chloroethanol utilizer pseudomonas sp. strain gj1, leading to a new steady state in which about 1% of the mix ...19938250561
development of field application vectors for bioremediation of soils contaminated with polychlorinated biphenyls.field application vectors (favs), which are a combination of a selective substrate, a host, and a cloning vector, have been developed for the purpose of expressing foreign genes in nonsterile, competitive environments in which the gene products provide no advantage to the host. such gene products are exemplified by the enzymes for the cometabolism of polychlorinated biphenyls (pcbs) through the biphenyl degradation pathway. attempts to use highly competent pcb-cometabolizing strains in the envir ...19938328798
oxidation of polychlorinated biphenyls by pseudomonas sp. strain lb400 and pseudomonas pseudoalcaligenes kf707.biphenyl-grown cells and cell extracts prepared from biphenyl-grown cells of pseudomonas sp. strain lb400 oxidize a much wider range of chlorinated biphenyls than do analogous preparations from pseudomonas pseudoalcaligenes kf707. these results are attributed to differences in the substrate specificity of the biphenyl 2,3-dioxygenases from both organisms.19938331086
new aerobic benzoate oxidation pathway via benzoyl-coenzyme a and 3-hydroxybenzoyl-coenzyme a in a denitrifying pseudomonas sp.a denitrifying pseudomonas sp. is able to oxidize aromatic compounds compounds completely to co2, both aerobically and anaerobically. it is shown that benzoate is aerobically oxidized by a new degradation pathway via benzoyl-coenzyme a (coa) and 3-hydroxybenzoyl-coa. the organism grew aerobically with benzoate, 3-hydroxybenzoate, and gentisate; catechol, 2-hydroxybenzoate, and protocatechuate were not used, and 4-hydroxybenzoate was a poor substrate. mutants were obtained which were not able to ...19938335640
biodegradation of 4-nitrotoluene by pseudomonas sp. strain 4nt.a strain of pseudomonas spp. was isolated from nitrobenzene-contaminated soil on 4-nitrotoluene as the sole source of carbon, nitrogen, and energy. the organism also grew on 4-nitrobenzaldehyde, and 4-nitrobenzoate. 4-nitrobenzoate and ammonia were detected in the culture fluid of glucose-grown cells after induction with 4-nitrotoluene. washed suspensions of 4-nitrotoluene- or 4-nitrobenzoate-grown cells oxidized 4-nitrotoluene, 4-nitrobenzaldehyde, 4-nitrobenzyl alcohol, and protocatechuate. ex ...19938357257
oxidation of biphenyl by a multicomponent enzyme system from pseudomonas sp. strain lb400.pseudomonas sp. strain lb400 grows on biphenyl as the sole carbon and energy source. this organism also cooxidizes several chlorinated biphenyl congeners. biphenyl dioxygenase activity in cell extract required addition of nad(p)h as an electron donor for the conversion of biphenyl to cis-2,3-dihydroxy-2,3-dihydrobiphenyl. incorporation of both atoms of molecular oxygen into the substrate was shown with 18o2. the nonlinear relationship between enzyme activity and protein concentration suggested t ...19938419290
purification and properties of the physically associated meta-cleavage pathway enzymes 4-hydroxy-2-ketovalerate aldolase and aldehyde dehydrogenase (acylating) from pseudomonas sp. strain cf600.the final two steps in the dmp operon-encoded meta-cleavage pathway for phenol degradation in pseudomonas sp. strain cf600 involve conversion of 4-hydroxy-2-ketovalerate to pyruvate and acetyl coenzyme a (acetyl-coa) by the enzymes 4-hydroxy-2-ketovalerate aldolase and aldehyde dehydrogenase (acylating) [acetaldehyde:nad+ oxidoreductase (coa acetylating), ec 1.2.1.10]. a procedure for purifying these two enzyme activities to homogeneity is reported here. the two activities were found to copurify ...19938419288
effects of medium and trace metals on kinetics of carbon tetrachloride transformation by pseudomonas sp. strain kc.under denitrifying conditions, pseudomonas sp. strain kc transforms carbon tetrachloride (ct) to carbon dioxide via a complex but as yet undetermined mechanism. transformation rates were first order with respect to ct concentration over the ct concentration range examined (0 to 100 micrograms/liter) and proportional to protein concentration, giving pseudo-second-order kinetics overall. addition of ferric iron (1 to 20 microm) to an actively transforming culture inhibited ct transformation, and t ...19938357248
degradation of 2-chloroallylalcohol by a pseudomonas sp.three pseudomonas strains capable of utilizing 2-chloroallylalcohol (2-chloropropenol) as the sole carbon source for growth were isolated from soil. the fastest growth was observed with strain jd2, with a generation time of 3.6 h. degradation of 2-chloroallylalcohol was accompanied by complete dehalogenation. chloroallylalcohols that did not support growth were dechlorinated by resting cells; the dechlorination level was highest if an alpha-chlorine substituent was present. crude extracts of str ...19938434917
characterization of the structural gene encoding a copper-containing nitrite reductase and homology of this gene to dna of other denitrifiers.a copper-containing nitrite reductase gene (niru) from pseudomonas sp. strain g-179 was found in a 1.9-kb ecori-bamhi dna fragment. the coding region contained information for a polypeptide of 379 amino acids. the encoded protein had 78% identity in amino acid sequence to the nitrite reductase purified from achromobacter cycloclastes. the ligands for type 1 copper- and type 2 copper-binding sites found in a. cycloclastes were also found in pseudomonas sp. strain g-179, suggesting that these bind ...19938439151
cloning and characterization of pseudomonas sp. strain dnt genes for 2,4-dinitrotoluene degradation.the degradation of 2,4-dinitrotoluene (dnt) by pseudomonas sp. strain dnt is initiated by a dioxygenase attack to yield 4-methyl-5-nitrocatechol (mnc) and nitrite. subsequent oxidation of mnc by a monooxygenase results in the removal of the second molecule of nitrite, and further enzymatic reactions lead to ring fission. initial studies on the molecular basis of dnt degradation in this strain revealed the presence of three plasmids. mitomycin-derived mutants deficient in either dnt dioxygenase o ...19938449889
metabolism of 2,2'-dihydroxybiphenyl by pseudomonas sp. strain hbp1: production and consumption of 2,2',3-trihydroxybiphenyl.cells of pseudomonas sp. strain hbp1 grown on 2-hydroxy- or 2,2'-dihydroxybiphenyl contain nadh-dependent monooxygenase activity that hydroxylates 2,2'-dihydroxybiphenyl. the product of this reaction was identified as 2,2',3-trihydroxybiphenyl by 1h nuclear magnetic resonance and mass spectrometry. furthermore, the monooxygenase activity also hydroxylates 2,2',3-trihydroxybiphenyl at the c-3' position, yielding 2,2',3,3'-tetrahydroxybiphenyl as a product. an estradiol ring cleavage dioxygenase a ...19938449871
cloning and nucleotide sequence of the gene encoding the positive regulator (dmpr) of the phenol catabolic pathway encoded by pvi150 and identification of dmpr as a member of the ntrc family of transcriptional activators.the catabolic plasmid pvi150 of pseudomonas sp. strain cf600 encodes all the genetic information required for the regulated metabolism of phenol and some of its methyl-substituted derivatives. the structural dmp genes of the pathway are clustered in a single operon that lies just downstream of a -24 tggc, -12 ttgc nif/ntr-like promoter sequence. promoters of this class are recognized by a minor form of rna polymerase utilizing sigma 54 (ntra, rpon). primer extension analysis demonstrated that th ...19938449869
selection of pseudomonas sp. strain hbp1 prp for metabolism of 2-propylphenol and elucidation of the degradative pathway.a mutant of pseudomonas sp. strain hbp1, originally isolated on 2-hydroxybiphenyl, was selected for the ability to grow on 2-propylphenol as the sole carbon and energy source. in the mutant strain, which was designated as pseudomonas sp. strain hbp1 prp, the cellular induction mechanism involved in the synthesis of the nadh-dependent monooxygenase is changed. 2-propylphenol, which is known to be a substrate of the monooxygenase, does not induce formation of the monooxygenase in the wild type but ...19938481010
construction of a pseudomonas hybrid strain that mineralizes 2,4,6-trinitrotoluene.a bacterium, pseudomonas sp. strain c1s1, able to grow on 2,4,6-trinitrotoluene (tnt), 2,4- and 2,6-dinitrotoluene, and 2-nitrotoluene as n sources, was isolated. the bacterium grew at 30 degrees c with fructose as a c source and accumulated nitrite. through batch culture enrichment, we isolated a derivative strain, called pseudomonas sp. clone a, which grew faster on tnt and did not accumulate nitrite in the culture medium. use of tnt by these two strains as an n source involved the successive ...19938468288
n.m.r. spectroscopic studies of fucose-containing oligosaccharides derived from keratanase digestion of articular cartilage keratan sulphates. influence of fucose residues on keratanase cleavage.keratan sulphate chains from bovine articular cartilage were fully digested with keratanase from pseudomonas sp. and the products were reduced with alkaline borohydride. the resultant fragments were fractionated on a nucleosil 5sb column and the earliest eluting fucose-containing oligosaccharides were isolated. structural analysis using 1h n.m.r. spectroscopy (600 mhz) showed the two least-charged species to have the following structure: glcnac(6s) beta 1-3gal beta 1-4(fuc alpha 1-3)glcnac(6s) b ...19938489515
developmental regulation of the gene for formate dehydrogenase in neurospora crassa.we have isolated and characterized a gene, fdh, from neurospora crassa which is developmentally regulated and which produces formate dehydrogenase activity when expressed in escherichia coli. the gene is closely linked (less than 0.6 kb apart) to the leu-5 gene encoding mitochondrial leucyl-trna synthetase; the two genes are transcribed convergently from opposite strands. the expression patterns of these genes differ: fdh mrna is found only during conidiation and early germination and is not det ...19938509325
microbial oxidation of dimethylnaphthalene isomers.three bacterial strains, identified as alcaligenes sp. strain d-59 and pseudomonas sp. strains d-87 and d-186, capable of growing on 2,6-dimethylnaphthalene (2,6-dmn) as the sole source of carbon and energy were isolated from soil samples. 2,6-naphthalene dicarboxylic acid was formed in the culture broths of these three strains grown on 2,6-dmn. in addition, 2-hydroxymethyl-6-methylnaphthalene and 6-methylnaphthalene-2-carboxylic acid were detected in the culture broth of strain d-87. strain d-8 ...19938517744
physiological factors affecting carbon tetrachloride dehalogenation by the denitrifying bacterium pseudomonas sp. strain kc.pseudomonas sp. strain kc was grown on a medium with a low content of transition metals in order to examine the conditions for carbon tetrachloride (ct) transformation. several carbon sources, including acetate, glucose, glycerol, and glutamate, were able to support ct transformation. the chelators 2,2'-dipyridyl and 1,10-phenanthroline stimulated ct transformation in a rich medium that otherwise did not support this activity. low (< 10 microm) additions of dissolved iron(ii), iron(iii), and cob ...19938517754
maleylacetate reductases in chloroaromatic-degrading bacteria using the modified ortho pathway: comparison of catalytic properties.the maleylacetate reductases from pseudomonas aeruginosa rho1 and alcaligenes eutrophus jmp134 were tested for activity and affinity to various maleylacetates as well as dechlorinating properties. the dechlorinating activity and the kcat/km values revealed high-level similarity of these reductases to that of pseudomonas sp. strain b13.19968550433
degradation of chloroaromatics: purification and characterization of maleylacetate reductase from pseudomonas sp. strain b13.maleylacetate reductase of pseudomonas sp. strain b13 was purified to homogeneity by chromatography on deae-cellulose, butyl-sepharose, blue-sepharose, and sephacryl s100. the final preparation gave a single band by polyacrylamide gel electrophoresis under denaturing conditions and a single symmetrical peak by gel filtration under nondenaturing conditions. the subunit m(r) value was 37,000 (determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis). estimation of the native m(r) va ...19938407778
a novel dipeptidyl aminopeptidase from pseudomonas sp. strain wo24.an activity similar to that of dipeptidyl aminopeptidase i (dap i) which releases dipeptide from gly-arg-p-nitroanilide (gly-arg-pna) was detected in a pseudomonas sp. an enzyme was isolated and purified about 400-fold by a series of column chromatographies. the enzyme, named dap bi (dap from bacteria, type i), was revealed to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page) and isoelectric focusing. the molecular mass was estimated to be 82 kda by sds-page ...19968631703
catechol 2,3-dioxygenases functional in oxygen-limited (hypoxic) environments.we studied the degradation of toluene for bacteria isolated from hypoxic (i.e., oxygen-limited) petroleum-contaminated aquifers and compared such strains with other toluene degraders. three pseudomonas isolates, p. pickettii pko1, pseudomonas sp. strain w31, and p. fluorescens cfs215, grew on toluene when nitrate was present as an alternate electron acceptor in hypoxic environments. we examined kinetic parameters (k(m) and vmax) for catechol 2,3-dioxygenase (c230), a key shared enzyme of the tol ...19968633871
stereospecific dihydroxylation of the styrene vinyl group by purified naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4.naphthalene dioxygenase (ndo) from pseudomonas sp. strain ncib 9816-4 adds both atoms of the dioxygen molecule to styrene to form (r)-l-phenyl-1,2-ethanediol. product formation is tightly coupled to dioxygen consumption and nadh oxidation. ndo oxidizes styrene-d8 at almost the same initial rate as styrene. the results indicate that dioxygen activation by ndo is different from that by cytochrome p-450 and other monooxygenases, which oxidize styrene to styrene 1,2-oxide.19968655521
atrazine chlorohydrolase from pseudomonas sp. strain adp: gene sequence, enzyme purification, and protein characterization.pseudomonas sp. strain adp metabolizes atrazine to carbon dioxide and ammonia via the intermediate hydroxyatrazine. the genetic potential to produce hydroxyatrazine was previously attributed to a 1.9-kb avai dna fragment from strain adp (m. l. de souza, l. p. wackett, k. l. boundy-mills, r. t. mandelbaum, and m. j. sadowsky, appl. environ. microbiol. 61:3373-3378, 1995). in this study, sequence analysis of the 1.9-kb avai fragment indicated that a single open reading frame, atza, encoded an acti ...19968759853
genetic structures of the genes encoding 2,3-dihydroxybiphenyl 1,2-dioxygenase and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid hydrolase from biphenyl- and 4-chlorobiphenyl-degrading pseudomonas sp. strain dj-12.the pcbc and pcbd genes of pseudomonas sp. strain dj-12, a natural isolate degrading biphenyl and 4-chlorobiphenyl, encode the 2,3-dihydroxybiphenyl 1,2-dioxygenase and 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid hydrolase, respectively. the two genes were sequenced and appear to be present in the order pcbd-pcbc as an operon.19968572703
chloroform mineralization by toluene-oxidizing bacteria.seven toluene-oxidizing bacterial strains (pseudomonas mendocina kr1, burkholderia cepacia g4, pseudomonas putida f1, pseudomonas pickettii pko1, and pseudomonas sp. strains envpc5, envbf1, and env113) were tested for their ability to degrade chloroform (cf). the greatest rate of cf oxidation was achieved with strain envbf1 (1.9 nmol/min/mg of cell protein). cf also was oxidized by p. mendocina kr1 (0.48 nmol/min/mg of cell protein), strain envpc5 (0.49 nmol/min/mg of cell protein), and escheric ...19968702263
phenotypic and genotypic characterization of phenanthrene-degrading fluorescent pseudomonas biovars.a total of 41 phenanthrene degraders were isolated from a former coal gasification site by using pseudomonas-selective gould's s1 medium. all isolates were found to belong to the fluorescent pseudomonas group and were subjected to characterization by phenotypic methods, including classical taxonomic tests, api 20ne, and biolog gn, and the strains were further characterized by the genotypic method repetitive extragenic palindromic pcr (rep-pcr). by using classical tests, the population was found ...19968837438
genetic exchange in soil between introduced chlorobenzoate degraders and indigenous biphenyl degraders.pseudomonas aeruginosa jb2, a chlorobenzoate degrader, was inoculated into soil having indigenous biphenyl degraders but no identifiable 2-chlorobenzoate (2cba) or 2,5-dichlorobenzoate (2,5dcba) degraders. the absence of any indigenous chlorobenzoate degraders was noted by the failure to obtain enrichment cultures with the addition of 2cba, 3cba, or 2,5dcba and by the failure of soil dna to hybridize to the tfdc gene, which encodes ortho fission of chlorocatechols. in contrast, dna extracted fro ...19968837452
cloning, sequencing, and expression of isopropylbenzene degradation genes from pseudomonas sp. strain jr1: identification of isopropylbenzene dioxygenase that mediates trichloroethene oxidation.pseudomonas sp. strain jr1, recently isolated with isopropylbenzene (ipb) as the inducer substrate for trichloroethene (tce) oxidation (b. dabrock, j. riedel, j. bertram, and g. gottschalk, arch. microbiol 158:9-13, 1992), is able to degrade ipb via the meta-cleavage pathway. the genes encoding the first three enzymes in the catabolism of isopropylbenzene were isolated from a genomic library with the broad-host-range cosmid vector pwe15. a 7.6-kb fragment from a 37.7-kb primary cosmid clone was ...19968899984
toluene and ethylbenzene oxidation by purified naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4.purified naphthalene dioxygenase (ndo) from pseudomonas sp. strain ncib 9816-4 oxidized toluene to benzyl alcohol and benzaldehyde by reactions involving benzylic monooxygenation and dioxygen-dependent alcohol oxidation, respectively. xylene and nitrotoluene isomers were also oxidized to substituted benzyl alcohol and benzaldehyde derivatives. ndo oxidized ethylbenzene sequentially through (s)-1-phenethyl alcohol (77% enantiomeric excess) and acetophenone to 2-hydroxyacetophenone. in addition, n ...19968795196
regio- and stereospecific oxidation of fluorene, dibenzofuran, and dibenzothiophene by naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4.the regio- and stereospecific oxidation of fluorene, dibenzofuran, and dibenzothiophene was examined with mutant and recombinant strains expressing naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4. the initial oxidation products were isolated and identified by gas chromatography-mass spectrometry and nuclear magnetic resonance spectrometry. salicylate-induced cells of pseudomonas sp. strain 9816/11 and isopropyl-beta-d-thiogalactopyranoside-induced cells of escherichia coli jm109( ...19968899998
oxidation of 6,7-dihydro-5h-benzocycloheptene by bacterial strains expressing naphthalene dioxygenase, biphenyl dioxygenase, and toluene dioxygenase yields homochiral monol or cis-diol enantiomers as major products.bacterial strains expressing naphthalene, biphenyl, and toluene dioxygenase were examined for their abilities to oxidize 6,7-dihydro-5h-benzocycloheptene (benzocyclohept-1-ene). the major oxidation products were isolated, and their absolute configurations were determined by chiral 1h nuclear magnetic resonance analysis of diastereomeric boronate esters, chiral stationary-phase high-pressure liquid chromatography, and stereo-chemical correlation. pseudomonas sp. strain 9816/11 and sphingomonas ya ...19968919798
measurement of minimum substrate concentration (smin) in a recycling fermentor and its prediction from the kinetic parameters of pseudomonas strain b13 from batch and chemostat cultures.the minimum substrate concentration required for growth, smin, was measured for pseudomonas sp. strain b13 with 3-chlorobenzoate (3cb) and acetate in a recycling fermentor. the substrates were provided alone or in a mixture. smin values predicted with kinetic parameters from resting-cell batches and chemostat cultures differed clearly from the values measured in the recycling fermentor. when 3cb and acetate were fed as single substrates, the measured smin values were higher than the individual s ...19968967775
comparison of factors influencing trichloroethylene degradation by toluene-oxidizing bacteria.the degradation of trichloroethylene (tce) by toluene-oxidizing bacteria has been extensively studied, and yet the influence of environmental conditions and physiological characteristics of individual strains has received little attention. to consider these effects, the levels of tce degradation by strains distinguishable on the basis of toluene and nitrate metabolism were compared under aerobic or hypoxic conditions in the presence and absence of nitrate and an exogenous electron donor, lactate ...19968975612
purification of an extracellular d-(-)-3-hydroxybutyrate oligomer hydrolase from pseudomonas sp. strain a1 and cloning and sequencing of its gene.an extracellular d-(-)-3-hydroxybutyrate oligomer hydrolase was purified from a poly(3-hydroxybutyrate)-degrading bacterium, pseudomonas sp. strain a1. the purified enzyme hydrolyzed the d-(-)-3-hydroxybutyrate dimer and trimer at similar rates. the enzyme activity was inhibited by a low concentration of diisopropylfluorophosphate. the molecular weight of the hydrolase was estimated to be about 70,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. a 10-kbp dna fragment of a1 was d ...19978981982
enzymology of oxidation of tropic acid to phenylacetic acid in metabolism of atropine by pseudomonas sp. strain at3.pseudomonas sp. strain at3 grew with dl-tropic acid, the aromatic component of the alkaloid atropine, as the sole source of carbon and energy. tropic acid-grown cells rapidly oxidized the growth substrate, phenylacetaldehyde, and phenylacetic acid. crude cell extracts, prepared from dl-tropic acid-grown cells, contained two nad+-linked dehydrogenases which were separated by ion-exchange chromatography and shown to be specific for their respective substrates, dl-tropic acid and phenylacetaldehyde ...19979023182
the atzb gene of pseudomonas sp. strain adp encodes the second enzyme of a novel atrazine degradation pathway.we previously reported the isolation of a 21.5-kb genomic dna fragment from pseudomonas sp. strain adp, which contains the atza gene, encoding the first metabolic step for the degradation of the herbicide atrazine (m. de souza, l. p. wackett, k. l. boundy-mills, r. t. mandelbaum, and m. j. sadowsky, appl. environ. microbiol. 61:3373-3378, 1995). in this study, we show that this fragment also contained the second gene of the atrazine metabolic pathway, atzb. atzb catalyzed the transformation of h ...19979055410
two types of novel dipeptidyl aminopeptidases from pseudomonas sp. strain wo24.two kinds of dipeptidyl aminopeptidase i (dap i [cathepsin c])-like activities which hydrolyze gly-phe-p-nitroanilide (gly-phe-pna) were detected in pseudomonas sp. strain wo24. they were purified and characterized. the isolated enzymes, named dap bii and dap biii, were revealed to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (sds-page) and isoelectric focusing. dap bii was estimated to have a molecular mass of 150,000 da by gel filtration and a subunit size of 73, ...19968892831
heterologous expression of biphenyl dioxygenase-encoding genes from a gram-positive broad-spectrum polychlorinated biphenyl degrader and characterization of chlorobiphenyl oxidation by the gene products.the bpha1a2a3a4 gene cluster, encoding a biphenyl dioxygenase from rhodococcus globerulus p6, a gram-positive microorganism able to degrade a wide spectrum of polychlorobiphenyls (pcbs), was expressed in pseudomonas putida, thereby allowing characterization of chlorobiphenyl oxidation by this enzyme. while p6 biphenyl dioxygenase activity was observed in p. putida containing bpha1a2a3a4, no activity was detected in escherichia coli cells containing the same gene cluster. in e. coli, transcriptio ...19979068637
molecular characterization of genes of pseudomonas sp. strain hr199 involved in bioconversion of vanillin to protocatechuate.the gene loci vdh, vana, and vanb, which are involved in the bioconversion of vanillin to protocatechuate by pseudomonas sp. strain hr199 (dsm 7063), were identified as the structural genes of a novel vanillin dehydrogenase (vdh) and the two subunits of a vanillate demethylase (vana and vanb), respectively. these genes were localized on an ecori fragment (e230), which was cloned from a pseudomonas sp. strain hr199 genomic library in the cosmid pvk100. the vdh gene was identified on a subfragment ...19979098058
preparation of gm1 ganglioside with sialidase-producing marine bacteria as a microbial biocatalyst.this paper describes the preparation of monosialoganglioside gm1 with sialidase-producing marine bacteria as a microbial biocatalyst. a new sialidase-producing bacterium, identified tentatively as pseudomonas sp. strain yf-2, was isolated from seawater by enrichment culture with ganglioside as the sole source of carbon. when yf-2 was cultured in a synthetic medium containing crude bovine brain gangliosides at 25 degrees c for 3 days, 80 to 90% of the gangliosides were converted to gm1. gm1 was t ...19979143118
stereospecific oxidation of (r)- and (s)-1-indanol by naphthalene dioxygenase from pseudomonas sp. strain ncib 9816-4.a recombinant escherichia coli strain which expresses naphthalene dioxygenase (ndo) from pseudomonas sp. strain ncib 9816-4 oxidized (s)-1-indanol to trans-(1s,3s)-indan-1,3-diol (95.5%) and (r)-3-hydroxy-1-indanone (4.5%). the same cells oxidized (r)-1-indanol to cis-1,3-indandiol (71%), (r)-3-hydroxy-1-indanone (18.2%), and cis-1,2,3-indantriol (10.8%). purified ndo oxidized (s)-1-indenol to both syn- and anti-2,3-dihydroxy-1-indanol.19979143136
structure and expression of a pyrimidine gene cluster from the extreme thermophile thermus strain zo5.on a 4.7-kbp hindiii clone of thermus strain zo5 dna, complementing an aspartate carbamoyltransferase mutation in escherichia coli, we identified a cluster of four potential open reading frames corresponding to genes pyrr, and pyrb, an unidentified open reading frame named bbc, and gene pyrc. the transcription initiation site was mapped at about 115 nucleotides upstream of the pyrr translation start codon. the cognate thermus pyr promoter also functions in heterologous expression of thermus pyr ...19979171389
cloning, characterization, and sequence analysis of the clce gene encoding the maleylacetate reductase of pseudomonas sp. strain b13.a 3,167-bp psti fragment of genomic dna from pseudomonas sp. strain b13 was cloned and sequenced. the gene clce consists of 1,059 nucleotides encoding a protein of 352 amino acids with a calculated mass of 37,769 da which showed maleylacetate reductase activity. the protein had significant sequence similarities with the polypeptides encoded by tcbf of pp51 (59.4% identical positions), tfdf of pjp4 (55.1%), and tfte of burkholderia cepacia ac1100 (53.1%). the function of tcbf as maleylacetate red ...19979171435
gene cloning, sequence analysis, and expression of 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase.the gene encoding 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase (mhpco; ec 1.14.12.4) was cloned by using an oligonucleotide probe corresponding to the n terminus of the enzyme to screen a dna library of pseudomonas sp. ma-1. the gene encodes for a protein of 379 amino acid residues corresponding to a molecular mass of 41.7 kda, the same as that previously estimated for mhpco. mhpco was expressed in escherichia coli and found to have the same properties as the native enzyme from pseudom ...19979207074
bacterial dl-2-haloacid dehalogenase from pseudomonas sp. strain 113: gene cloning and structural comparison with d- and l-2-haloacid dehalogenases.dl-2-haloacid dehalogenase from pseudomonas sp. strain 113 (dl-dex) catalyzes the hydrolytic dehalogenation of both d- and l-2-haloalkanoic acids to produce the corresponding l- and d-2-hydroxyalkanoic acids, respectively, with inversion of the c2 configuration. dl-dex is a unique enzyme: it acts on the chiral carbon of the substrate and uses both enantiomers as equivalent substrates. we have isolated and sequenced the gene encoding dl-dex. the open reading frame consists of 921 bp corresponding ...19979209038
cloning of genes involved in carbazole degradation of pseudomonas sp. strain ca10: nucleotide sequences of genes and characterization of meta-cleavage enzymes and hydrolase.the dna fragment encoding meta-cleavage enzymes and the meta-cleavage compound hydrolase, involved in carbazole degradation, was cloned from the carbazole-utilizing bacterium pseudomonas sp. strain ca10. dna sequence analysis of this 2.6-kb smai-sphi fragment revealed that there were three open reading frames (orf1, orf2, and orf3, in this gene order). orf1 and orf2 were indispensable for meta-cleavage activity for 2'-aminobiphenyl-2,3-diol and its easily available analog, 2,3-dihydroxybiphenyl, ...19979244273
identification and characterization of genes encoding carbazole 1,9a-dioxygenase in pseudomonas sp. strain ca10.nucleotide sequence analysis of the flanking regions of the carbc genes of pseudomonas sp. strain ca10 revealed that there were two open reading frames (orfs) orf4 and orf5, in the upstream region of carbc. similarly, three orfs, orf6 to orf8, were found in the downstream region of carbc. the deduced amino acid sequences of orf6 and orf8 showed homologies with ferredoxin and ferredoxin reductase components of bacterial multicomponent dioxygenase systems, respectively. orf4 and orf5 had the same ...19979244274
use of a promoterless lacz gene insertion to investigate chitinase gene expression in the marine bacterium pseudoalteromonas sp. strain s9.sequence data for genes encoding 16s rrna indicated that the marine strain previously named pseudomonas sp. strain s9 would be better identified as a pseudoalteromonas sp. by use of transposon mutagenesis, a chitinase-negative mutant of s9 with a lacz reporter gene insertion was isolated. part of the interrupted gene was cloned and sequenced. the deduced amino acid sequence had homology to sequences of bacterial chitinases. expression of the chitinase gene promoter was quantified by measuring th ...19979251187
multiple pathways for toluene degradation in burkholderia sp. strain js150.burkholderia (pseudomonas) sp. strain js150 uses multiple pathways for the metabolism of catechols that result from degradation of aromatic compounds. this suggests that the strain also uses multiple upstream pathways for the initial hydroxylation of aromatic substrates. two distinct dna fragments that allowed pseudomonas aeruginosa pao1c to grow with benzene as a sole carbon source were cloned from strain js150. one of the recombinant plasmids containing the initial steps for the degradative pa ...19979327568
chemotaxis of pseudomonas spp. to the polyaromatic hydrocarbon naphthalene.two naphthalene-degrading bacteria, pseudomonas putida g7 and pseudomonas sp. strain ncib 9816-4, were chemotactically attracted to naphthalene in drop assays and modified capillary assays. growth on naphthalene or salicylate induced the chemotactic response. p. putida g7 was also chemotactic to biphenyl; other polyaromatic hydrocarbons that were tested did not appear to be chemoattractants for either pseudomonas strain. strains that were cured of the naphthalene degradation plasmid were not att ...19979327579
cross-regulation of toluene monooxygenases by the transcriptional activators tbmr and tbut.the toluene-3-monooxygenase from burkholderia pickettii pko1 and the toluene/benzene-2-monooxygenase from burkholderia (pseudomonas) sp. strain js150 are distinct enzymes which differ not only in catalytic specificity and substrate range but also in the arrangement and sequence of the genes within the operons that encode the enzymes, tbua1ubva2c and tbmabcdef, respectively. in the present study, we examined the transcriptional activation of the ptbua1 and ptbma promoters by their cognate regulat ...19979293027
microbiologically proven bacterial infections in aids.we have reviewed the incidence, type and site of microbiologically proven bacterial infection occurring in 52 patients with the acquired immunodeficiency syndrome (aids) who presented to southmead hospital, bristol between 1990 and 1994. a total of 30 (58%) patients had significant bacterial isolates. the majority of infections were community acquired. overall, more infections were caused by gram-negative organisms but gram-positive organisms predominated in bacteraemia. mycobacterium avium intr ...19979373597
atzc is a new member of the amidohydrolase protein superfamily and is homologous to other atrazine-metabolizing enzymes.pseudomonas sp. strain adp metabolizes atrazine to cyanuric acid via three plasmid-encoded enzymes, atza, atzb, and atzc. the first enzyme, atza, catalyzes the hydrolytic dechlorination of atrazine, yielding hydroxyatrazine. the second enzyme, atzb, catalyzes hydroxyatrazine deamidation, yielding n-isopropylammelide. in this study, the third gene in the atrazine catabolic pathway, atzc, was cloned from a pseudomonas sp. strain adp cosmid library as a 25-kb ecori dna fragment in escherichia coli. ...19989422605
comparative in vitro activities of trovafloxacin (cp-99,219) against 221 aerobic and 217 anaerobic bacteria isolated from patients with intra-abdominal infections.four hundred thirty-eight bacteria cultured from specimens of patients with serious intra-abdominal infections were tested by agar dilution against trovafloxacin and other quinolones and antimicrobial agents. trovafloxacin inhibited 435 strains (99.3%) at < or =2 microg/ml. all the quinolones had similar activities against enterobacteriaceae and pseudomonas sp., but trovafloxacin showed superior activities against streptococci, enterococci, and anaerobic organisms. because of its excellent in vi ...19979333074
a cold-adapted lipase of an alaskan psychrotroph, pseudomonas sp. strain b11-1: gene cloning and enzyme purification and characterization.a psychrotrophic bacterium producing a cold-adapted lipase upon growth at low temperatures was isolated from alaskan soil and identified as a pseudomonas strain. the lipase gene (lipp) was cloned from the strain and sequenced. the amino acid sequence deduced from the nucleotide sequence of the gene (924 bp) corresponded to a protein of 308 amino acid residues with a molecular weight of 33,714. lipp also has consensus motifs conserved in other cold-adapted lipases, i.e., lipase 2 from antarctic m ...19989464382
enzyme specificity of 2-nitrotoluene 2,3-dioxygenase from pseudomonas sp. strain js42 is determined by the c-terminal region of the alpha subunit of the oxygenase component.biotransformations with recombinant escherichia coli expressing the genes encoding 2-nitrotoluene 2,3-dioxygenase (2ntdo) from pseudomonas sp. strain js42 demonstrated that 2ntdo catalyzes the dihydroxylation and/or monohydroxylation of a wide range of aromatic compounds. extremely high nucleotide and deduced amino acid sequence identity exists between the components from 2ntdo and the corresponding components from 2,4-dinitrotoluene dioxygenase (2,4-dntdo) from burkholderia sp. strain dnt (form ...19989495758
genetic and functional analysis of the styrene catabolic cluster of pseudomonas sp. strain y2.the chromosomal region of pseudomonas sp. strain y2 involved in the conversion of styrene to phenylacetate (upper catabolic pathway) has been cloned and sequenced. four catabolic genes, styabcd, and two regulatory genes, stysr, were identified. this gene cluster when transferred to escherichia coli w confers to this phenylacetate-degrading host the ability to grow on styrene as the sole carbon and energy source. genes styabcd are homologous to those encoding the styrene upper catabolic pathway i ...19989495743
anti-mitochondrial antibodies in patients with dilated cardiomyopathy (anti-m7) are directed against flavoenzymes with covalently bound fad.anti-mitochondrial antibodies (anti-m7) in sera from patients with dilated cardiomyopathy and myocarditis recognize, besides mitochondrial antigens, bacterial sarcosine dehydrogenase. the common target antigen was identified as the covalently bound fad of mitochondrial and bacterial flavoenzymes. thus, anti-m7-positive serum reacted on western blots exclusively with covalently flavinylated enzymes. the antigenic specificity of anti-m7 sera was reproduced by an antiserum raised in rabbits with 6- ...19989528896
ntn genes determining the early steps in the divergent catabolism of 4-nitrotoluene and toluene in pseudomonas sp. strain tw3.pseudomonas sp. strain tw3 is able to oxidatively metabolize 4-nitrotoluene and toluene via a route analogous to the upper pathway of the tol plasmids. we report the sequence and organization of five genes, ntnwcmab*, which are very similar to and in the same order as the xyl operon of tol plasmid pww0 and present evidence that they encode enzymes which are expressed during growth on both 4-nitrotoluene and toluene and are responsible for their oxidation to 4-nitrobenzoate and benzoate, respecti ...19989555884
a gene cluster encoding steps in conversion of naphthalene to gentisate in pseudomonas sp. strain u2.pseudomonas sp. strain u2 was isolated from oil-contaminated soil in venezuela by selective enrichment on naphthalene as the sole carbon source. the genes for naphthalene dioxygenase were cloned from the plasmid dna of strain u2 on an 8.3-kb bamhi fragment. the genes for the naphthalene dioxygenase genes nagaa (for ferredoxin reductase), nagab (for ferredoxin), and nagac and nagad (for the large and small subunits of dioxygenase, respectively) were located by southern hybridizations and by nucle ...19989573207
low-frequency horizontal transfer of an element containing the chlorocatechol degradation genes from pseudomonas sp. strain b13 to pseudomonas putida f1 and to indigenous bacteria in laboratory-scale activated-sludge microcosms.the possibilities for low-frequency horizontal transfer of the self-transmissible chlorocatechol degradative genes (clc) from pseudomonas sp. strain b13 were investigated in activated-sludge microcosms. when the clc genes were transferred into an appropriate recipient bacterium such as pseudomonas putida f1, a new metabolic pathway for chlorobenzene degradation was formed by complementation which could be selected for by the addition of mono- or 1, 4-dichlorobenzene (cb). under optimized conditi ...19989603824
the atzabc genes encoding atrazine catabolism are located on a self-transmissible plasmid in pseudomonas sp. strain adp.pseudomonas sp. strain adp initiates atrazine catabolism via three enzymatic steps, encoded by atza, -b, and -c, which yield cyanuric acid, a nitrogen source for many bacteria. in-well lysis, southern hybridization, and plasmid transfer studies indicated that the atza, -b, and -c genes are localized on a 96-kb self-transmissible plasmid, padp-1, in pseudomonas sp. strain adp. high-performance liquid chromatography analyses showed that cyanuric acid degradation was not encoded by padp-1. padp-1 w ...19989603862
towards a biocatalyst for (s)-styrene oxide production: characterization of the styrene degradation pathway of pseudomonas sp. strain vlb120.in order to design a biocatalyst for the production of optically pure styrene oxide, an important building block in organic synthesis, the metabolic pathway and molecular biology of styrene degradation in pseudomonas sp. strain vlb120 was investigated. a 5.7-kb xhoi fragment, which contained on the same strand of dna six genes involved in styrene degradation, was isolated from a gene library of this organism in escherichia coli by screening for indigo formation. t7 rna polymerase expression expe ...19989603811
detoxification of protoanemonin by dienelactone hydrolase.protoanemonin is a toxic metabolite which may be formed during the degradation of some chloroaromatic compounds, such as polychlorinated biphenyls, by natural microbial consortia. we show here that protoanemonin can be transformed by dienelactone hydrolase of pseudomonas sp. strain b13 to cis-acetylacrylate. although similar km values were observed for cis-dienelactone and protoanemonin, the turnover rate of protoanemonin was only 1% that of cis-dienelactone. this indicates that at least this pe ...19989440530
utility of gram's stain and efficacy of quantitative cultures for posttraumatic pneumonia: a prospective study.this prospective trial examined the efficacy of using bronchoalveolar lavage (bal) for the diagnosis of pneumonia (pn) and the utility of gram's stain (gs) for dictating empiric therapy.19989605666
chromosomal integration, tandem amplification, and deamplification in pseudomonas putida f1 of a 105-kilobase genetic element containing the chlorocatechol degradative genes from pseudomonas sp. strain b13.analysis of chlorobenzene-degrading transconjugants of pseudomonas putida f1 which had acquired the genes for chlorocatechol degradation (clc) from pseudomonas sp. strain b13 revealed that the clc gene cluster was present on a 105-kb amplifiable genetic element (named the clc element). in one such transconjugant, p. putida rr22, a total of seven or eight chromosomal copies of the entire genetic element were present when the strain was cultivated on chlorobenzene. chromosomal integrations of the ...19989721270
purification and characterization of the coniferyl aldehyde dehydrogenase from pseudomonas sp. strain hr199 and molecular characterization of the gene.the coniferyl aldehyde dehydrogenase (caldh) of pseudomonas sp. strain hr199 (dsm7063), which catalyzes the nad+-dependent oxidation of coniferyl aldehyde to ferulic acid and which is induced during growth with eugenol as the carbon source, was purified and characterized. the native protein exhibited an apparent molecular mass of 86,000 +/- 5,000 da, and the subunit mass was 49.5 +/- 2.5 kda, indicating an alpha2 structure of the native enzyme. the optimal oxidation of coniferyl aldehyde to feru ...19989721273
pseudomonas sp. strain 273, an aerobic alpha, omega-dichloroalkanedegrading bacterium.a gram-negative, aerobic bacterium was isolated from soil; this bacterium grew in 50% (vol/vol) suspensions of 1,10-dichlorodecane (1,10-dcd) as the sole source of carbon and energy. phenotypic and small-subunit ribosomal rna characterizations identified the organism, designated strain 273, as a member of the genus pseudomonas. after induction with 1,10-dcd, pseudomonas sp. strain 273 released stoichiometric amounts of chloride from c5 to c12 alpha, omega-dichloroalkanes in the presence of oxyge ...19989726906
identification and characterization of is1411, a new insertion sequence which causes transcriptional activation of the phenol degradation genes in pseudomonas putida.a new insertion sequence (is element), is1411, was identified downstream of the phenol degradation genes pheba that originated from plasmid dna of pseudomonas sp. strain est1001. according to sequence analysis, is1411 belongs to a new family of is elements that has recently been named the isl3 family (j. mahillon and m. chandler, microbiol. mol. biol. rev. 62:725-774, 1998). is1411 generates 8-bp duplication of the target dna and carries 24-bp inverted repeats (irs), highly homologous to the irs ...19989765560
int-b13, an unusual site-specific recombinase of the bacteriophage p4 integrase family, is responsible for chromosomal insertion of the 105-kilobase clc element of pseudomonas sp. strain b13.pseudomonas sp. strain b13 carries the clcrabde genes encoding chlorocatechol-degradative enzymes on the self-transmissible 105-kb clc element. the element integrates site and orientation specifically into the chromosomes of various bacterial recipients, with a glycine trna structural gene (glyv) as the integration site. we report here the localization and nucleotide sequence of the integrase gene and the activity of the integrase gene product in mediating site-specific integration. the integras ...19989791097
biochemical and genetic characterization of trans-2'-carboxybenzalpyruvate hydratase-aldolase from a phenanthrene-degrading nocardioides strain.trans-2'-carboxybenzalpyruvate hydratase-aldolase was purified from a phenanthrene-degrading bacterium, nocardioides sp. strain kp7, and characterized. the purified enzyme was found to have molecular masses of 38 kda by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 113 kda by gel filtration chromatography. thus, the homotrimer of the 38-kda subunit constituted an active enzyme. the km and kcat values of this enzyme for trans-2'-carboxybenzalpyruvate were 50 microm and 13 s(-1), r ...19989473051
construction and use of an ipb dna module to generate pseudomonas strains with constitutive trichloroethene and isopropylbenzene oxidation activity.pseudomonas sp. strain jr1 exhibits trichloroethene (tce) oxidation activity with isopropylbenzene (ipb) as the inducer substrate. we previously reported the genes encoding the first three enzymes of the ipb-degradative pathway (ipba1, ipba2, ipba3, ipba4, ipbb, and ipbc) and identified the initial ipb dioxygenase (ipba1 a2a3a4) as responsible for tce cooxidation (u. pflugmacher, b. averhoff, and g. gottschalk, appl. environ. microbiol. 62:3967-3977, 1996). primer extension analyses revealed mul ...19989647815
cloning of a sphingomonas paucimobilis syk-6 gene encoding a novel oxygenase that cleaves lignin-related biphenyl and characterization of the enzyme.sphingomonas paucimobilis syk-6 transforms 2,2'-dihydroxy-3,3'-dimethoxy-5,5'-dicarboxybiphenyl (ddva), a lignin-related biphenyl compound, to 5-carboxyvanillic acid via 2,2',3-trihydroxy-3'-methoxy-5,5'-dicarboxybiphenyl (oh-ddva) as an intermediate (15). the ring fission of oh-ddva is an essential step in the ddva degradative pathway. a 15-kb ecori fragment isolated from the cosmid library complemented the growth deficiency of a mutant on oh-ddva. subcloning and deletion analysis showed that a ...19989647824
molecular cloning, expression and site-directed mutagenesis of glutathione s-transferase from ochrobactrum anthropi.the gene coding for a novel glutathione s-transferase (gst) has been isolated from the bacterium ochrobactrum anthropi. a pcr fragment of 230 bp was obtained using oligonucleotide primers deduced from n-terminal and 'internal' sequences of the purified enzyme. the gene was obtained by screening of a genomic dna partial library from o. anthropi constructed in pbluescript with a pcr fragment probe. the gene encodes a protein (oagst) of 201 amino acids with a calculated molecular mass of 21738 da. ...19989794797
cloning and molecular analysis of the poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) biosynthesis genes in pseudomonas sp. strain 61-3.two types of polyhydroxyalkanoate (pha) biosynthesis gene loci (phb and pha) of pseudomonas sp. strain 61-3, which produces a blend of poly(3-hydroxybutyrate) [p(3hb)] homopolymer and a random copolymer (poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) [p(3hb-co-3ha]) consisting of 3ha units of 4 to 12 carbon atoms, were cloned and analyzed at the molecular level. in the phb locus, three open reading frames encoding polyhydroxybutyrate (phb) synthase (phbcps), beta-ketothiolase (phbaps), and nadph- ...19989851987
involvement of the terminal oxygenase beta subunit in the biphenyl dioxygenase reactivity pattern toward chlorobiphenyls.biphenyl dioxygenase (bph dox) oxidizes biphenyl on adjacent carbons to generate 2,3-dihydro-2,3-dihydroxybiphenyl in comamonas testosteroni b-356 and in pseudomonas sp. strain lb400. the enzyme comprises a two-subunit (alpha and beta) iron sulfur protein (ispbph), a ferredoxin (ferbph), and a ferredoxin reductase (redbph). b-356 bph dox preferentially catalyzes the oxidation of the double-meta-substituted congener 3,3'-dichlorobiphenyl over the double-para-substituted congener 4,4'-dichlorobiph ...19989811638
shewanella putrefaciens mtrb encodes an outer membrane protein required for fe(iii) and mn(iv) reduction.iron and manganese oxides or oxyhydroxides are abundant transition metals, and in aquatic environments they serve as terminal electron acceptors for a large number of bacterial species. the molecular mechanisms of anaerobic metal reduction, however, are not understood. shewanella putrefaciens is a facultative anaerobe that uses fe(iii) and mn(iv) as terminal electron acceptors during anaerobic respiration. transposon mutagenesis was used to generate mutants of s. putrefaciens, and one such mutan ...19989829939
genetic and biochemical analyses of the tec operon suggest a route for evolution of chlorobenzene degradation genes.the teca broad-spectrum chlorobenzene dioxygenase of burkholderia sp. strain ps12 catalyzes the first step in the mineralization of 1,2,4, 5-tetrachlorobenzene. the catabolic genes were localized on a small plasmid that belongs to the incpbeta incompatibility group. pcr analysis of the genetic environment of the tec genes indicated high similarity to the transposon-organized catabolic tcb chlorobenzene degradation genes of pseudomonas sp. strain p51. sequence analysis of the regions flanking the ...19999864349
thermal gradient gel electrophoresis analysis of bioprotection from pollutant shocks in the activated sludge microbial community.we used a culture-independent approach, namely, thermal gradient gel electrophoresis (tgge) analysis of ribosomal sequences amplified directly from community dna, to determine changes in the structure of the microbial community following phenol shocks in the highly complex activated sludge ecosystem. parallel experimental model sewage plants were given shock loads of chlorinated and methylated phenols and simultaneously were inoculated (i) with a genetically engineered microorganism (gem) able t ...19999872766
polycyclic aromatic hydrocarbon degradation by a new marine bacterium, neptunomonas naphthovorans gen. nov., sp. nov.two strains of bacteria were isolated from creosote-contaminated puget sound sediment based on their ability to utilize naphthalene as a sole carbon and energy source. when incubated with a polycyclic aromatic hydrocarbon (pah) compound in artificial seawater, each strain also degraded 2-methylnaphthalene and 1-methylnaphthalene; in addition, one strain, nag-2n-113, degraded 2,6-dimethylnaphthalene and phenanthrene. acenaphthene was not degraded when it was used as a sole carbon source but was d ...19999872786
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