Publications

cytochrome p450cam: crystallography, oxygen activation, and electron transfer.	several crystal structures of various substrate and inhibited complexes of the camphor monoxygenase, cytochrome p450cam from pseudomonas putida, are now available. these structures, together with mutagenesis, biochemical, and biophysical studies, have allowed for a detailed penetration into the problem of how p450s activate molecular oxygen, control stereoselectivity, and transfer electrons. this review will provide a summary of the crystallographic work in light of what these structures have ta ...	1992	1537455
identification of a new gene, tmof, in the pseudomonas mendocina kr1 gene cluster encoding toluene-4-monooxygenase.	five genes, tmoabcde, encoding toluene-4-monooxygenase (t4mo) were previously mapped to a 3.6-kb region of a 10.2-kb saci dna fragment isolated from pseudomonas mendocina kr1 (k.-m. yen, m. r. karl, l. m. blatt, m. j. simon, r. b. winter, p. r. fausset, h. s. lu, a. a. harcourt, and k. k. chen, j. bacteriol. 173:5315-5327, 1991). in this report, we describe the identification and characterization of a dna region in the saci fragment whose expression enhances the t4mo activity determined by the t ...	1992	1429451
characterization of catechol 2,3-dioxygenases.	three catechol 2,3-dioxygenases for biphenyl, naphthalene/salicylate, and toluene/xylene oxidation were cloned from achromobacter xylosoxidans kf701, pseudomonas putida (nah7), and pseudomonas sp. (pwwo). the cloned catechol 2,3-dioxygenases were identified by enzymatic activity assay in addition to yellow bands on polyacrylamide gel after electrophoresis and activity staining. all of the cloned catechol 2,3-dioxygenases exhibited their highest activities on catechol as a substrate compared with ...	1992	1543511
the influence of temperature on the behaviour of mixed bacterial contamination of the shell membrane of the hen's egg.	the inner membrane of the air cell of hens' eggs was inoculated with pseudomonas putida, staphylococcus xylosus, enterococcus faecalis, escherichia coli and salmonella enteritidis. the first mentioned eventually dominated the contamination of the albumen of eggs stored at 4, 15, and 20 degrees c. the last mentioned did so in eggs stored at 37 degrees c. the interval between inoculation of the membrane and gross contamination of the albumen was markedly influenced by site of contamination relativ ...	1992	1547832
effect of pseudobactin 358 production by pseudomonas putida wcs358 on suppression of fusarium wilt of carnations by nonpathogenic fusarium oxysporum fo47.	nonpathogenic fusarium oxysporum fo47b10 combined with pseudomonas putida wcs358 efficiently suppressed fusarium wilt of carnations grown in soilless culture. this suppression was significantly higher than that obtained by inoculation of either antagonistic microorganism alone. the increased suppression obtained by fo47b10 combined with wcs358 only occurred when fo47b10 was introduced at a density high enough (at least 10 times higher than that of the pathogen) to be efficient on its own. p. put ...	1992	1444411
degradation of the herbicide bromoxynil in pseudomonas putida.	biological conversion of the herbicide bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) was studied in a batch culture of pseudomonas putida by using hplc. the process had a cometabolic character and proceeded only in the presence of another, simultaneously metabolizable, carbon and energy source. the intensity of degradation correlated with the growth rate, the degradation stopping when the cosubstrate becomes exhausted or the ph value of the medium falls below 6.5. in a medium with glucose, no l ...	1992	1505868
cloning and partial sequencing of an operon encoding two pseudomonas putida haloalkanoate dehalogenases of opposite stereospecificity.	we have cloned fragments of dna (up to 13 kb), from pseudomonas putida aj1, that code for two stereospecific haloalkanoate dehalogenases. these enzymes are highly specific for d and l substrates. the two genes, designated hadd and hadl, have been isolated and independently expressed in escherichia coli and p. putida hosts by using broad-host-range vectors. they are closely adjacent and inducible in what appears to be an operon with an upstream open reading frame of unknown function. nucleotide s ...	1992	1556080
effect of glucose and ribose on microbial degradation of the herbicide bromoxynil continuously added to soil.	bromoxynil (3,5-dibromo-4-hydroxybenzonitrile) was continuously added to chernozem (haplic typic) soil inoculated with a suspension of pseudomonas putida capable of cometabolic decomposition of the compound in a hetero-continuous-flow cultivation setup. in the steady state, when glucose or ribose were simultaneously added, 90 and 47% of the added herbicide was degraded per day, respectively. if the saccharides were absent, only 10-27% of the herbicide was decomposed. addition and removal of gluc ...	1992	1505869
microbial oxidation of adamantanone by pseudomonas putida carrying the camphor catabolic plasmid.	intact cells of (+/-)camphor-grown pseudomonas putida, atcc17453(cam), have been shown to oxidize readily the monoketone derivative of cage hydrocarbon adamantane, forming oxygenated products indicative of both biological baeyer-villiger and hydroxylation reactions. formed products were identified as 4-oxahomoadamantan-5-one, 5-hydroxyadamantan-2-one and 1-hydroxy-4-oxahomoadamantan-5-one. minor products formed as a result of secondary reactions were tentatively identified as syn- and anti-1,4-d ...	1992	1510672
[the effect of physicochemical factors on the growth of pseudomonas putida bs-2 on a medium with diethylene glycol].	the physicochemical factors of medium have been studied for their effect on the physiological indices of growth of pseudomonas putida bs-2 culture utilizing diethylenglycol as the only source of carbon. action of the supraoptimal temperature on the growth process of p. putida bs-2 is accompanied by a decrease (more than twice) in economic coefficient of substrate and specific growth rate as compared with their maximal values. dependences of specific growth rate of p. putida bs-2 in the medium wi ...	1992	1406384
functional domains of aromatase cytochrome p450 inferred from comparative analyses of amino acid sequences and substantiated by site-directed mutagenesis experiments.	several functional domains, especially the active site regions, in aromatase cytochrome p450 were inferred by alignment of amino acid sequences of the enzyme from five species, human, rat, mouse, chicken, and trout, and that of pseudomonas putida cytochrome p450cam, whose x-ray structure has been determined (poulos, t.l., finzel, b.c., and howard, a.j. (1987) j. mol. biol. 195, 687-700). the predicted functions of these domains have been evaluated by site-directed mutagenesis. eighteen mutants, ...	1992	1429608
physiological properties of a pseudomonas strain which grows with p-xylene in a two-phase (organic-aqueous) medium.	pseudomonas putida idaho utilizes toluene, m-xylene, p-xylene, 1,2,4-trimethylbenzene, and 3-ethyltoluene as growth substrates when these hydrocarbons are provided in a two-phase system at 5 to 50% (vol/vol). growth also occurs on luria-bertani medium in the presence of a wide range of organic solvents. the ability of the organism to grow in the presence of organic solvents is correlated with the logarithm of the octanol-water partition coefficient, with dimethyl-phthalate (log p(oct) = 2.3) bei ...	1992	1444381
genetic variants in the putidaredoxin-cytochrome p-450cam electron-transfer complex: identification of the residue responsible for redox-state-dependent conformers.	camphor is hydroxylated in pseudomonas putida by a three-component system comprised of an oxidase, cytochrome p-450cam, and a two-protein electron-transfer chain, putidaredoxin and putidaredoxin reductase [tyson et al. (1972) j. biol. chem. 274, 5777-5784]. the enzymatic removal of putidaredoxin's c-terminal tryptophan is known to cause a much reduced rate of enzymatic activity in the reconstituted camphor hydroxylase system [sligar et al. (1974) proc. natl. acad. sci. u.s.a. 71, 3906-3910]. to ...	1992	1445875
roles of catr and cis,cis-muconate in activation of the catbc operon, which is involved in benzoate degradation in pseudomonas putida.	in pseudomonas putida, the catbc operon encodes enzymes involved in benzoate degradation. previous studies have determined that these enzymes are induced when p. putida is grown in the presence of benzoate. induction of the enzymes of the catbc operon requires an intermediate of benzoate degradation, cis,cis-muconate, and a regulatory protein, catr. it has been determined that catr binds to a 27-bp region of the catbc promoter in the presence or absence of inducer. we have called this the repres ...	1992	1447146
construction of a broad host range shuttle vector for gene cloning and expression in actinobacillus pleuropneumoniae and other pasteurellaceae.	we have constructed a pair of broad host range expression vectors, pjff224-nx and pjff224-xn, based on plasmid rsf1010, which enable cloning and efficient expression of genes in actinobacillus pleuropneumoniae and pasteurella haemolytica and in escherichia coli. the vectors consist of the minimal autonomous replicon of the broad host range plasmid rsf1010 and a type ii chloramphenicol acetyl transferase gene for chloramphenicol resistance selection. in addition, they contain a gene expression ca ...	1992	1448612
analysis of mutations in trfa, the replication initiation gene of the broad-host-range plasmid rk2.	plasmids with mutations in trfa, the gene encoding the replication initiation protein of the broad-host-range plasmid rk2, were isolated and characterized. mutants identified from a nitrosoguanidine bank were defective in supporting the replication of a wild-type rk2 origin in escherichia coli. most of the mutations were clustered in a region of trfa corresponding to the carboxy-terminal quarter of the trfa protein. 5' and 3' deletion mutants of trfa were also constructed. a c-terminal deletion ...	1992	1597426
dna sequence determination and functional characterization of the oct-plasmid-encoded alkjkl genes of pseudomonas oleovorans.	the alkbfghjkl and alkst operons encode enzymes that allow pseudomonas putida (oleovorans) to metabolize alkanes. in this paper we report the nucleotide sequence of a 4592 bp region of the alkbfghjkl operon encoding the alkj, alkk and alkl polypeptides. the alkj gene encodes a protein of 59 kilodaltons. the predicted amino acid sequence shows significant homology with four flavin proteins: choline dehydrogenase, a glucose dehydrogenase and two oxidases. alkj is membrane-bound and converts alipha ...	1992	1453953
expression of the 4-chlorobenzoate dehalogenase genes from pseudomonas sp.-cbs3 in escherichia coli and identification of the gene translation products.	the genes encoding the 4-chlorobenzoate dehalogenase of pseudomonas sp. strain cbs3 were, in an earlier study, cloned in escherichia coli dh1 with the cosmid vector ppsa843 and then mobilized to the 4-chlorobenzoate dehalogenase minus strain pseudomonas putida kt2440. in this paper we report on the expression of 4-chlorobenzoate dehalogenase in these clones and on the polypeptide composition of the active enzyme. the dehalogenase activity in whole cells suspended in 3.2 mm 4-chlorobenzoate (30 d ...	1992	1477786
evidence that the hrpb gene encodes a positive regulator of pathogenicity genes from pseudomonas solanacearum.	the hrp gene cluster of pseudomonas solanacearum gmi1000 strain encodes functions that are essential for pathogenicity on tomato and for the elicitation of the hypersensitive response on tobacco. in this study, we present the nucleotide sequence of one of the hrp genes (hrpb) located at the left-hand end of the cluster and we show that hrpb encodes a positive regulator controlling the expression of hrp genes. hrpb has a coding capacity for a 477-amino-acid polypeptide, which shows significant si ...	1992	1479894
multiple periplasmic catalases in phytopathogenic strains of pseudomonas syringae.	phytopathogenic strains of pseudomonas syringae are exposed to plant-produced, detrimental levels of hydrogen peroxide during invasion and colonization of host plant tissue. when p. syringae strains were investigated for their capacity to resist h2o2, they were found to contain 10- to 100-fold-higher levels of total catalase activity than selected strains belonging to nonpathogenic related taxa (pseudomonas fluorescens and pseudomonas putida) or escherichia coli. multiple catalase activities wer ...	1992	1514792
[electron-conformational interactions at the active site of reduced bacterial cytochrome p450cam induced by a substrate and analysis of the electron structure of heme].	magnetic circular dichroism (mcd) spectra in the soret region (360-480 nm) of camphor-free and camphor-bound reduced bacterial cytochrome p450cam from pseudomonas putida were recorded and analysed in the temperature range from 2 k to 290 k. the temperature dependences of the mcd intensity are qualitatively changed by binding of substrate to the enzyme. in the absence of camphor the linear increase of the mcd intensity with 1/t at t < 4.2 k gives evidence for degeneracy or near degeneracy of the ...	1992	1491671
genetic analysis of the agga locus involved in agglutination and adherence of pseudomonas putida, a beneficial fluorescent pseudomonad.	an isolate of pseudomonas putida, which rapidly adheres to plant roots, is agglutinated by a glycoprotein from root surfaces. agglutination is prevented and adherence to the root surface is diminished by tn5 insertion in mutant 5123. two cosmid clones from wild type p. putida and a 2.7-kbp ecori-hindiii subclone present in both cosmid clones restored agglutinable to wild type levels in transconjugants of the nonagglutinable (agg-) tn5 mutant 5123. these three clones increased agglutinability in ...	1992	1617198
[the kinetics of glycol destruction by a pseudomonas putida bs-2 strain].	destruction of ethylene glycol and diethylene glycol by pseudomonas putida bs-2 culture under conditions of its batch cultivation has been studied for its physiological regularities. the specific rate of the biomass growth in the region of limiting substrate concentrations depends on the diethylene glycol concentration in the medium and follows the mono equation. a semisaturation constant for diethylene glycol is 209 +/- 17 mg/d. the specific rate of the culture growth is independent of the ethy ...	1992	1453991
acyloin formation by benzoylformate decarboxylase from pseudomonas putida.	whole cells and cell extracts of pseudomonas putida grown in a medium containing ammonium mandelate have the capacity to produce the acyloin compound 2-hydroxypropiophenone when incubated with benzoylformate and acetaldehyde. benzaldehyde and benzyl alcohol were formed as reaction by-products. the enantiomeric excess of the 2-hydroxypropiophenone product was found to be 91 to 92%. the absolute configuration of the enzymatically prepared product at the carbinol carbon was found to be s. the thiam ...	1992	1622241
adaptation of model genetically engineered microorganisms to lake water: growth rate enhancements and plasmid loss.	when a genetically engineered microorganism (gem) is released into a natural ecosystem, its survival, and hence its potential environmental impact, depends on its genetic stability and potential for growth under highly oligotrophic conditions. in this study, we compared plasmid stability and potential for growth on low concentrations of organic nutrients of strains of pseudomonas putida serving as model gems. plasmid-free and plasmid-bearing (nah7) prototrophic isogenic strains and two amino-aci ...	1992	1482185
oxidation of nitrotoluenes by toluene dioxygenase: evidence for a monooxygenase reaction.	pseudomonas putida f1 and pseudomonas sp. strain js150 initiate toluene degradation by incorporating molecular oxygen into the aromatic nucleus to form cis-1,2-dihydroxy-3-methylcyclohexa-3,5-diene. when toluene-grown cells were incubated with 2- and 3-nitrotoluene, the major products identified were 2- and 3-nitrobenzyl alcohol, respectively. the same cells oxidized 4-nitrotoluene to 2-methyl-5-nitrophenol and 3-methyl-6-nitrocatechol. escherichia coli jm109(pdtg601), which contains the toluene ...	1992	1514810
dna homology between siderophore genes from fluorescent pseudomonads.	many species of pseudomonads produce fluorescent siderophores involved in iron uptake. we have investigated the dna homology between the siderophore synthesis genes of an opportunist animal pathogen, pseudomonas aeruginosa, and three plant-associated species pseudomonas syringae, pseudomonas putida and pseudomonas sp. b10. there is extensive homology between the dna from the different species, consistent with the suggestion that the different siderophore synthesis genes have evolved from the sam ...	1992	1532617
microbial metabolism of quinoline and related compounds. xiv. purification and properties of 1h-3-hydroxy-4-oxoquinoline oxygenase, a new extradiol cleavage enzyme from pseudomonas putida strain 33/1.	1h-3-hydroxy-4-oxoquinoline oxygenase was purified to apparent homogeneity from pseudomonas putida strain 33/1 which can use 1h-4-oxoquinoline as sole source of carbon. the molecular mass of the enzyme was determined to 26,000 da by gel chromatography and by sds polyacrylamide gel electrophoresis. the enzyme is labile at temperatures above 30 degrees c and has a ph optimum of 8.0. it requires oxygen for the reaction and is significantly inhibited by metal ions like cu2+, zn2+, hg2+ and by 4-chlo ...	1992	1515060
molecular cloning and characterization of catechol 2,3-dioxygenases from biphenyl/polychlorinated biphenyls-degrading bacteria.	catechol 2,3-dioxygenases were cloned from alcaligenes sp. kf711, pseudomonas putida kf715, and achromobacter xylosoxidans kf701 which are biphenyl/polychlorinated biphenyls-degrading bacteria. all of the cloned enzymes were purified by preparative polyacrylamide gel electrophoresis (page). the purified catechol 2,3-dioxygenases were significantly different from one another in ring-fission activities to catechol and its derivatives. the catechol 2,3-dioxygenase from alcaligenes sp. kf711 exhibit ...	1992	1530619
sequence and complementation analysis of recf genes from escherichia coli, salmonella typhimurium, pseudomonas putida and bacillus subtilis: evidence for an essential phosphate binding loop.	we have compared the recf genes from escherichia coli k-12, salmonella typhimurium, pseudomonas putida, and bacillus subtilis at the dna and amino acid sequence levels. to do this we determined the complete nucleotide sequence of the recf gene from salmonella typhimurium and we completed the nucleotide sequence of recf gene from pseudomonas putida begun by fujita et al. (1). we found that the recf proteins encoded by these two genes contain respectively 92% and 38% amino acid identity with the e ...	1992	1542576
lack of homology between two haloacetate dehalogenase genes encoded on a plasmid from moraxella sp. strain b.	two genes encoding haloacetate dehalogenases, h-1 and h-2, are closely linked on a plasmid from moraxella sp. strain b. h-1 predominantly acts on fluoroacetate, but h-2 does not. to elucidate the molecular relationship between the two enzymes, we compared their structural genes. two restriction fragments of the plasmid dna were subcloned on m13 phages and their nucleotide sequences were determined. the sequence of each fragment contained an open reading frame that was identified as the structura ...	1992	1512562
molecular characterization of the entner-doudoroff pathway in escherichia coli: sequence analysis and localization of promoters for the edd-eda operon.	the nucleotide sequence of the entire escherichia coli edd-eda region that encodes the enzymes of the entner-doudoroff pathway was determined. the edd structural gene begins 236 bases downstream of zwf. the eda structural gene begins 34 bases downstream of edd. the edd reading frame is 1,809 bases long and encodes the 602-amino-acid, 64,446-da protein 6-phosphogluconate dehydratase. the deduced primary amino acid sequences of the e. coli and zymomonas mobilis dehydratase enzymes are highly conse ...	1992	1624451
purification of pro- and eukaryotic superoxide dismutases by charge-controlled hydrophobic chromatography.	the process of purifying superoxide dismutases was simplified using charge-controlled hydrophobic chromatography on 10-carboxydecyl sepharose. in only one chromatographic step following ammonium sulphate precipitation, fe-containing superoxide dismutase from pseudomonas putida and cu,zn-containing superoxide dismutase from bovine erythrocytes were purified with an overall yield of about 70% to electrophoretic homogeneity. the specific activities of the crystalline enzyme preparations were expres ...	1992	1560096
plasmids with easily excisable xyle cassettes.	two new vectors containing the xyle gene (encoding catechol-2,3-dioxygenase) of pseudomonas putida were constructed that serve as the source of the xyle cassette. these vectors are based on the kanamycin-resistance-encoding plasmid, pkan18. the promoter-less xyle gene is flanked by several restriction enzyme sites that allow for easy excision of this gene in the form of a cassette containing a ribosome-binding site, 7 bp upstream from the start codon. these cassettes lack any transcriptional ter ...	1992	1644310
nitroaromatics are substrates for the tol plasmid upper-pathway enzymes.	expression of the xylma genes encoding for toluene monoxygenase from the lactose promoter in a broad-host-range plasmid allows the oxidation of toluene and m- and p-nitrotoluene to their corresponding benzyl alcohols and benzaldehydes in pseudomonas putida and escherichia coli. benzyl alcohols accumulate until reaching a concentration of about 80 mum, while benzaldehydes accumulate steadily with time for at least 24 h. tol-encoded benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase recog ...	1992	16348637
differential siderophore utilization and iron uptake by soil and rhizosphere bacteria.	the differential availabilities of the hydroxamate siderophores ferrioxamine b (fob) and ferrichrome (fc) and the pseudobactin siderophores st3, 7nsk(2), and wcs 358 as sources of fe for soil and rhizosphere bacteria were studied. about 20% of the total bacterial cfu from the rhizospheres of four plant species were able to use fob as the sole fe source in an fe-deficient medium, while about 12, 10, 2, and > 1% were able to use fc and pseudobactins 7nsk(2), st3, and wcs 358, respectively. of the ...	1992	16348618
enhanced bacterial metabolism of a pseudomonas strain in response to the addition of culture filtrate of a bacteriophagous amoeba.	in a previous work, levrat et al. [21] showed an enhancement of the production of pyoverdin (siderophore) by pseudomonas putida in the presence of amoeba. to explain the mechanism of stimulation, the hypothesis of production of stimulatory factors by amoeba was proposed. filtrates of both mixed culture of bacteria and amoeba (pseudomonas putida + acanthamoeba castellanii) and of axenic culture of amoeba were added to the culture medium of pseudomonas. the production of pyoverdin was increased in ...	1992	23194985
a plant selectable marker gene based on the detoxification of the herbicide dalapon.	a gene from pseudomonas putida coding for a dehalogenase capable of degrading 2,2 dichloropropionic acid (2,2dcpa), the active ingredient of the herbicide dalapon, has been isolated and characterised. in plant transformation experiments the gene was shown to confer resistance to 2,2dcpa at a tissue culture level where 2,2dcpa could be used to select for transformants. at the whole plant level, transformed plants showed resistance to 2,2dcpa at concentrations up to 5 times the recommended dose ra ...	1992	24213366
use of an exotic carbon source to selectively increase metabolic activity and growth of pseudomonas putida in soil.	respiration and growth of pseudomonas putida ppg7, containing catabolic plasmid nah7, was determined in three agricultural field soils amended with the carbon source salicylate. the addition of salicylate to soil significantly increased the population of ppg7. however, there was a lack of relationship between microbial numbers and activity as determined by evolution of co(2). in soils containing 30 to 1,500 mug of salicylate per g, metabolic activities of ppg7 peaked between 18 and 42 h and popu ...	1993	16348983
enhanced growth and activity of a biocontrol bacterium genetically engineered to utilize salicylate.	plasmid nah7 was transferred from pseudomonas putida ppg7 to p. putida r20 [r20(nah7)], an antagonist of pythium ultimum. the plasmid did not affect growth or survival of r20(nah7) and was stably maintained under nonselective conditions in broth and soil and on sugar beet seeds. plasmid nah7 conferred to r20(nah7) the ability to utilize salicylate in culture, agricultural field soil, and on sugar beet seeds. the metabolic activity of r20(nah7), but not the wild-type r20, was greatly increased in ...	1993	16348985
enhancement of population densities of pseudomonas putida ppg7 in agricultural ecosystems by selective feeding with the carbon source salicylate.	sodium salicylate (1,000 mug/ml) was delivered through a drip irrigation system to agricultural field soils planted to tomato and infested with pseudomonas putida ppg7, the host of the salicylate catabolic plasmid nah7. in nonfumigated soils infested with approximately 10 cfu of ppg7 per g in the top 30 cm, population densities were increased up to 112-fold within 14 days of the initial application of salicylate compared with the densities in the respective nonamended soils. mean season-long pop ...	1993	16348984
methylmercury resistance in desulfovibrio desulfuricans strains in relation to methylmercury degradation.	two strains of desulfovibrio desulfuricans, one known to synthesize monomethylmercury from ionic mercury, were grown to determine methylmercury toxicity and for comparison with an anaerobic strain of clostridium pasteurianum, a h(2) producer, and with the broad-spectrum mercury-resistant pseudomonas putida strain fb-1, capable of degrading 1 mug of methylmercury to methane and elemental mercury in 2 h. the ch(3)hgcl resistance of d. desulfuricans strains was 10 times that of p. putida fb-1 and 1 ...	1993	16349013
stable tagging of rhizobium meliloti with the firefly luciferase gene for environmental monitoring.	a system for stable tagging of gram-negative bacteria with the firefly luciferase gene, luc, is described. a previously constructed fusion constitutively expressing luc from the lambdap(r) promoter was used. stable integration into the bacterial genome was achieved by use of mini-tn5 delivery vectors. the procedure developed was applied for tagging of representative gram-negative bacteria, such as escherichia coli, rhizobium meliloti, pseudomonas putida, and agrobacterium tumefaciens. the system ...	1993	16349015
application of a tetrazolium salt with a water-soluble formazan as an indicator of viability in respiring bacteria.	the tetrazolium salt sodium 3'-{1-[(phenylamino)-carbonyl]-3,4-tetrazolium}-bis (4-methoxy-6-nitro)benzene-sulfonic acid hydrate (xtt) was examined for use as a colorimetric indicator of viability in respiring bacteria. xtt was reduced to an orange, water-soluble formazan product by methylosinus trichosporium ob3b, pseudomonas putida, escherichia coli, and bacillus subtilis. formazan production was proportional to live cell biomass, and xtt was reduced by all cultures in the absence of added ele ...	1993	16349038
purification and characterization of an l-aminopeptidase from pseudomonas putida atcc 12633.	an l-aminopeptidase of pseudomonas putida, used in an industrial process for the hydrolysis of d,l-amino acid amide racemates, was purified to homogeneity. the highly l-enantioselective enzyme resembled thiol reagent-sensitive alkaline serine proteinases and was strongly activated by divalent cations. it possessed a high substrate specificity for dipeptides and alpha-h amino acid amides, e.g., l-phenylglycine amide.	1993	16349130
stereospecific hydroxylation of indan by escherichia coli containing the cloned toluene dioxygenase genes from pseudomonas putida f1.	[this corrects the article on p. 3407 in vol. 58.].	1993	16348858
antagonistic effect of nonpathogenic fusarium oxysporum fo47 and pseudobactin 358 upon pathogenic fusarium oxysporum f. sp. dianthi.	pseudobactin production by pseudomonas putida wcs358 significantly improves biological control of fusarium wilt caused by nonpathogenic fusarium oxysporum fo47b10 (p. lemanceau, p. a. h. m. bakker, w. j. de kogel, c. alabouvette, and b. schippers, appl. environ. microbiol. 58:2978-2982, 1992). the antagonistic effect of fo47b10 and purified pseudobactin 358 was studied by using an in vitro bioassay. this bioassay allows studies on interactions among nonpathogenic f. oxysporum fo47b10, pathogenic ...	1993	16348860
method for investigation of competition between bacteria as a function of three environmental factors varied simultaneously.	competition between microorganisms as affected by temperature, ph, and the sodium chloride (nacl) concentration was investigated by selective replication from gradient plates. salmonella typhimurium was inhibited by pseudomonas putida at 20 and 23 degrees c but not 30 and 35 degrees c. p. putida no longer grew at the extremes of ph and nacl concentration, particularly at 30 and 35 degrees c.	1993	16348978
induction and characterization of a cytochrome p-450-dependent camphor hydroxylase in tissue cultures of common sage (salvia officinalis).	(+)-camphor, a major monoterpene of the essential oil of common sage (salvia officinalis), is catabolized in senescent tissue, and the pathway for the breakdown of this bicyclic ketone has been previously elucidated in sage cell-suspension cultures. in the initial step of catabolism, camphor is oxidized to 6-exo-hydroxycamphor, and the corresponding nadph- and o2-dependent hydroxylase activity was demonstrated in microsomal preparations of sage cells. several well-established inhibitors of cytoc ...	1993	12231778
human glyoxalase i. cdna cloning, expression, and sequence similarity to glyoxalase i from pseudomonas putida.	glyoxalase i (ec 4.4.1.5) catalyzes the transformation of methylglyoxal and glutathione to s-lactoylglutathione. we have isolated human cdna clones encoding glyoxalase i from a phorbol myristate acetate-treated u937 cdna library. this cdna encodes a protein of 184 amino acids with a calculated m(r) of 20,719. the amino acid composition calculated from the deduced amino acid sequence agreed with that reported for glyoxalase i purified from human erythrocytes. the escherichia coli cells carrying t ...	1993	7684374
biological synthesis of the analgesic hydromorphone, an intermediate in the metabolism of morphine, by pseudomonas putida m10.	the morphine alkaloid hydromorphone (dihydromorphinone) was identified as an intermediary metabolite in the degradation of morphine by pseudomonas putida m10. a constitutive nadh-dependent morphinone reductase capable of catalyzing the reduction of the 7,8-unsaturated bond of morphinone and codeinone, yielding hydromorphone and hydrocodone, respectively, was shown to be present in cell extracts. the structures have been identified by 1h nuclear magnetic resonance and mass spectrometry. morphinon ...	1993	7689317
transcription of the cam operon and camr genes in pseudomonas putida ppg1.	in pseudomonas putida carrying the cam plasmid, the operon (camdcab) encoding enzymes involved in the degradation pathway of d-camphor is negatively regulated by the camr protein, and camr is autorepressed. s1 nuclease mapping revealed that camdcab and camr were divergently transcribed from overlapping promoters, the transcription start sites were separated by 11 bp, and transcriptions of the cam operon (camdcab) and camr increased about 10- and 4-fold, respectively, immediately after addition o ...	1993	7693653
polymerase chain reaction amplification of naphthalene-catabolic and 16s rrna gene sequences from indigenous sediment bacteria.	we report the amplification of bacterial genes from uninoculated surface and subsurface sediments by the polymerase chain reaction (pcr). pcr amplification of indigenous bacterial 16s ribosomal dna genes was unsuccessful when subsurface sediment containing approximately 10(7) cells.g-1 was added directly to a pcr mixture. however, when 10 mg of sediment was inoculated with approximately 10(5) cells of pseudomonas putida g7, the nahac naphthalene dioxygenase gene characteristic of the p. putida g ...	1993	7683182
cloning and sequencing of a dehalogenase gene encoding an enzyme with hydrolase activity involved in the degradation of gamma-hexachlorocyclohexane in pseudomonas paucimobilis.	in pseudomonas paucimobilis ut26, gamma-hexachlorocyclohexane (gamma-hch) is converted by two steps of dehydrochlorination to a chemically unstable intermediate, 1,3,4,6-tetrachloro-1,4-cyclohexadiene (1,4-tcdn), which is then metabolized to 2,5-dichloro-2,5-cyclohexadiene-1,4-diol (2,5-ddol) by two steps of hydrolytic dehalogenation via the chemically unstable intermediate 2,4,5-trichloro-2,5-cyclohexadiene-1-ol (2,4,5-dnol). to clone a gene encoding the enzyme responsible for the conversion of ...	1993	7691794
antimicrobial activity and biosynthesis of indole antibiotics produced by xenorhabdus nematophilus.	we have investigated the mechanism of action and physiology of production of the indole derivative antibiotics produced by the nematode-associated, entomopathogenic bacterium xenorhabdus nematophilus. maximum antibiotic concentration was reached during the late stationary phase of growth, and the antibiotic yield was appreciably enhanced by supplementation with tryptophan. antibiotic biosynthesis apparently involved the removal of the side-chain carboxyl (c-1) carbon of tryptophan. the c-3 methy ...	1993	7510325
survival and impact of genetically engineered pseudomonas putida harboring mercury resistance gene in aquatic microcosms.	the survival of wild-type and genetically engineered pseudomonas putida ppy101 that contained a recombinant plasmid psr134 conferring mercury resistance were monitored in aquatic microcosms. we used lake, river, and spring water samples. the density of genetically engineered and wild-type p. putida decreased rapidly within 5 days (population change rate k -0.87 approximately -1.00 day-1), then moderately after 5 to 28 days (-0.10 approximately -0.14 day-1). the population change rates of genetic ...	1993	7764012
aerobic, phenol-induced tce degradation in completely mixed, continuous-culture reactors.	both pseudomonas putida f1 and a mixed culture were used to study tce degradation in continuous culture under aerobic, non-methanotrophic conditions. tce mass balance studies were performed with continuous culture reactors to determine the total percent removed in the reactors, and to quantify the percent removed by air stripping and biodegradation. adsorption of tce to biomass was assumed to be negligible. this research demonstrated the feasibility of treating tce-contaminated water under aerob ...	1993	7763855
cosubstrate effects in reductive dehalogenation by pseudomonas putida g786 expressing cytochrome p-450cam.	cytochrome p-450cam was shown to be the primary catalyst mediating reductive dehalogenation of polychlorinated ethanes by pseudomonas putida g786. under anaerobic conditions, the enzyme catalyzed reductive elimination reactions in vivo with the substrates hexachloroethane, pentachloroethane, and 1,1,1,2-tetrachlorethane; the products were tetrachloroethylene, trichloroethylene, and 1,1-dichloroethylene, respectively. in vivo reaction rates were determined. no reaction was observed with 1,1,2,2-t ...	1993	7763853
transcriptional control of the pseudomonas putida tol plasmid catabolic pathways.	tol plasmid pww0 of pseudomonas putida contains two operons that specify a pathway for the degradation of aromatic hydrocarbons. the upper pathway operon encodes the enzymes for the oxidation of toluene/xylenes to benzoate/toluates, and the metacleavage pathway operon encodes the enzymes for the further oxidation of these compounds to krebs cycle intermediates. their expression is controlled by the gene products of two divergently transcribed regulatory genes, xyir and xyis. the xyir protein, wh ...	1993	7934920
in vitro and in situ survivals of bacterial populations added to fresh water environments.	the fate of aeromonas hydrophila, alcaligenes denitrificans, vibrio cholerae non-01, pseudomonas putida and four different isolates of escherichia coli in fresh river water were assessed by using different microcosms (i.e., membrane diffusion chamber and erlenmeyer flask). when water samples were incubated at 16 +/- 1 degrees c, the differences in extent of survival among test bacteria were in general not significant. if the incubation temperature was raised to 29 +/- 1 degrees c, in the in situ ...	1993	7982366
characterization of cell surface properties in agglutinable and nonagglutinable mutants of pseudomonas putida.	cells of an aggressive, root-colonizing isolate of pseudomonas putida are agglutinated by a root surface glycoprotein. the agglutination phenotype in p. putida isolate corvallis is lacking in mutants (agg-) derived by tn5 insertion and chemical mutagenesis. specific mutation in the agga locus by tn5 insertion results in loss of agglutinability that is complemented in trans by a wild-type copy of the p. putida agga locus. we examined the biochemical bases of agglutination in p. putida by comparin ...	1993	8106135
identification and characterization of a siderophore regulatory gene (pfra) of pseudomonas putida wcs358: homology to the alginate regulatory gene algq of pseudomonas aeruginosa.	genes encoding biosynthesis of pseudobactin 358 (a microbial iron transport agent) and its cognate outer membrane receptor protein, pupa, are transcribed only under iron limitation in plant growth-promoting pseudomonas putida wcs358. two cosmid clones were identified from a gene bank of wcs358 dna which could independently and in an iron-dependent manner activate transcription from a wcs358 siderophore gene promoter in heterologous pseudomonas strain a225. the functional region of one of the clo ...	1993	7968519
mineralization of p-methyl-14c-benzoate in soils by pseudomonas putida (pww0).	pseudomonas putida bearing the archetypal tol plasmid pww0 metabolizes p-methylbenzoate through a meta-cleavage pathway. in complex environments such as soils that are relatively rich in organic matter, we observed metabolism of p-methyl-14c-benzoate, which could be monitored as 14co2 evolution from the labelled alkylaromatic. linear 14co2 evolution in soils took place for at least a month, although efficient mineralization of the alkylaromatic required appropriate mass transfer to allow the bac ...	1993	8111534
intrinsic stability and extrinsic stabilization of creatinase from pseudomonas putida.	creatinase (creatine amidinohydrolase, ec 3.5.3.3), a homodimer of 45 kda subunit molecular mass, shows only limited functional stability, and is inaccessible to reconstitution after preceding deactivation, denaturation and dissociation. the enzyme has been characterized regarding its native and denatured states. studying its unfolding characteristics in the presence of "extrinsic factors", such as dte, bsa and glycerol, it was possible to define solvent conditions where the stability of the enz ...	1993	8216893
cloning, expression and mutational analysis of the urocanase gene (hutu) from pseudomonas putida.	the histidine-utilizing hutu gene was isolated from a lambda-embl3 phage of a genomic library from pseudomonas putida nicii and subcloned into the expression vector pt7-7. escherichia coli bl21 cells were transformed with the recombinant plasmid and produced a catalytically active protein, amounting to approximately 30% of the total protein in the crude cell-free extract. the addition of nad+ to the growth medium ensured the full occupation of active sites by the cofactor. this requires a mechan ...	1993	7901006
engineering enzymes for clinical diagnosis.	the enzyme creatine amidinohydrolase (creatinase, ec 3.5.3.3.) is a major limiting factor in the enzymatic creatinine determination because of its comparatively poor catalytic activity and stability in the native form. the gene from pseudomonas putida coding for creatinase was cloned and used for overexpression of the protein in e coli and pseudomonas. in addition, it was possible by means of 'random' mutagenesis in vivo and subsequent screening using an activity plate assay to isolate creatinas ...	1993	8166396
variation in chlorobenzoate catabolism by pseudomonas putida p111 as a consequence of genetic alterations.	pseudomonas putida p111 is able to utilize a broad range of monochlorinated, dichlorinated, and trichlorinated benzoates. the involvement of two separate dioxygenases was noted from data on plasmid profiles and dna hybridization. the benzoate dioxygenase, which converts 3-chlorobenzoate (3-cb), 4-cb, and benzoate to the corresponding catechols via reduction of a dihydrodiol, was shown to be chromosomally coded. the chlorobenzoate-1,2-dioxygenase that converts ortho-chlorobenzoates to the corresp ...	1993	8215353
stabilization of creatinase from pseudomonas putida by random mutagenesis.	creatinase (creatine amidinohydrolase, ec 3.5.3.3) from pseudomonas putida is a homodimer of 45 kda subunit molecular mass, the three-dimensional structure of which is known at 1.9 a resolution. three point mutants, a109v, v355m, and v182i, as well as one double mutant combining a109v and v355m, and the triple mutant with all three replacements, were compared with wild-type creatinase regarding their physical and enzymological properties. high-resolution crystal data for wild-type creatinase and ...	1993	8251936
evidence for autoregulation of camr, which encodes a repressor for the cytochrome p-450cam hydroxylase operon on the pseudomonas putida cam plasmid.	the regulatory gene camr on the cam plasmid of pseudomonas putida (atcc 17453) negatively controls expression of the cytochrome p-450cam hydroxylase operon (camdcab) for the camphor degradation pathway and is oriented in a direction opposite to that of the camdcab operon. in this study, we examined expression of the camr gene by monitoring the beta-galactosidase activity of camr-lacz translational fusions in p. putida camr and camr+ strains. we found that the camr gene was autogenously regulated ...	1993	8253671
new derivatives of tol plasmid pww0.	two new segregants, ppw1-1 and ppw161-1, of pseudomonas putida were isolated from the stock cultures paw85(pww0) and paw85(pww0-161). strain ppw1-1 had lost its ability to grow on m-xylene but was able to grow on m-toluate. a deletion of the left-hand of transposon tn4651, including the upper-operon genes, had taken place in plasmid pww0mut1, isolated from strain ppw1-1. additional deletions were observed in pww0mut1 after 'benzoate-curing' (plasmids pww0mut15, pww0mut19, pww0mut27). the genes o ...	1993	8254307
microbial metabolism of quinoline and related compounds. xix. degradation of 4-methylquinoline and quinoline by pseudomonas putida k1.	a bacterial strain, designated k1, which utilizes 4-methylquinoline and quinoline as sole source of carbon, nitrogen and energy was isolated from soil. based on its morphological and physiological characteristics, it was classified as pseudomonas putida biovar b. four metabolites of 4-methylquinoline degradation were isolated from the culture supernatant and identified as 4-methyl-2-oxo-1,2-dihydroquinoline, 8-hydroxy-4-methyl-2-oxo-1,2-dihydroquinoline, 7,8-dihydroxy-4-methyl-2-oxo-1,2-dihydroq ...	1993	8216899
adaptive mutation: the uses of adversity.	when populations of microorganisms are subjected to certain nonlethal selections, useful mutants arise among the nongrowing cells whereas useless mutants do not. this phenomenon, known as adaptive, directed, or selection-induced mutation, challenges the long-held belief that mutations only arise at random and without regard for utility. in recent years a growing number of studies have examined adaptive mutation in both bacteria and yeast. although conflicts and controversies remain, the weight o ...	1993	8257106
construction and use of a new broad-host-range lacz transcriptional fusion vector, phrp309, for gram- bacteria.	a new lacz transcriptional fusion vector, phrp309, based on the incq plasmid rsf1010, was constructed and shown to be easily mobilized into a variety of gram- eubacteria. we also developed a two-step cloning procedure to facilitate the cloning of small promoter fragments into the fusion vector. a set of 'cohort' vectors was constructed which allowed directed cloning of fragments downstream from an omega streptomycin/spectinomycin-resistance cassette while maintaining multiple flanking restrictio ...	1993	8224891
oxidation of carbazole to 3-hydroxycarbazole by naphthalene 1,2-dioxygenase and biphenyl 2,3-dioxygenase.	naphthalene 1,2-dioxygenase from pseudomonas sp. ncib 9816-4 and biphenyl dioxygenase from beijerinckia sp. b8/36 oxidized the aromatic n-heterocycle carbazole to 3-hydroxycarbazole. toluene dioxygenase from pseudomonas putida f39/d did not oxidize carbazole. transformations were carried out by mutant strains which oxidize naphthalene and biphenyl to cis-dihydrodiols, and with a recombinant e. coli strain expressing the structural genes of naphthalene 1,2-dioxgenase from pseudomonas sp. ncib 981 ...	1993	8270195
a novel structural basis for membrane association of a protein: construction of a chimeric soluble mutant of (s)-mandelate dehydrogenase from pseudomonas putida.	the (s)-mandelate dehydrogenase (mdh) from pseudomonas putida (atcc 12633) is the only membrane-associated member of a homologous family of fmn-dependent, alpha-hydroxy acid dehydrogenases/oxidases that includes the structurally characterized glycolate oxidase from spinach (gox). we have correlated the membrane association of mdh to a polypeptide segment in the interior of the primary sequence. this has been accomplished by construction of a chimeric enzyme in which the putative membrane-binding ...	1993	8241149
quinoline oxidoreductase from pseudomonas putida 86: an improved purification procedure and electron paramagnetic resonance spectroscopy.	quinoline oxidoreductase, an iron-sulfur molybdenum flavoprotein containing flavin adenine dinucleotide and molybdopterin cytosine dinucleotide, was purified from pseudomonas putida 86 to homogeneity. the various electron-transfer centers of the purified enzyme were examined by electron paramagnetic resonance spectroscopy. quinoline deuterated at position 2 was prepared by deuterodecarboxylation of 2-quinolinecarboxylic acid. quinoline added to the enzyme elicited the mo(v) "rapid" type q signal ...	1993	8251516
purification and characterization of pseudomonas putida histidine ammonia-lyase expressed in escherichia coli.	histidine ammonia-lyase (hal) from pseudomonas putida prs1 contains a catalytically important electrophilic center reported to be dehydroalanine. little is known about the origin of this group or its linkage to the protein. to initiate structural studies on this enzyme, p. putida hal was purified from an escherichia coli high-expression clone in which the hal gene (huth) was under the control of the lambda pl promoter on a plasmid vector. in this clone from 6 to 10% of the soluble cell protein a ...	1993	8251759
regulation of the catechol 1,2-dioxygenase- and phenol monooxygenase-encoding pheba operon in pseudomonas putida paw85.	in pseudomonas putida paw85, the ortho-cleavage pathway is used for catechol degradation. the 11.4-kb xhoi fragment cloned from phenol degradation plasmid pest1226 into pkt240 (recombinant plasmid pat1140) contains the inducible pheba operon that encodes catechol 1,2-dioxygenase (gene pheb) and phenol monooxygenase (gene phea), the first two enzymes for the phenol degradation pathway. the promoter of the pheba operon is mapped 1.5 kb upstream of the pheb gene. the plasmid pat1140, when introduce ...	1993	8253692
analysis of the mrna structure of the pseudomonas putida tol meta fission pathway operon around the transcription initiation point, the xylte and the xylfj regions.	the 13 genes encoded by the meta-cleavage operon (approx. 11 kb) of pseudomonas putida tol plasmid pww0 are transcribed from a single promoter, pm. in p. putida, transcription from pm was strictly dependent on the presence of effector-activated xyls protein. three regions of the transcript were analyzed in the wild-type strain of p. putida (pww0) by s1 nuclease protection and primer extension analyses. a major point of transcription initiation was found in the most 5'-end of the operon, which de ...	1993	8241263
complete nucleotide sequence of the 5-exo-hydroxycamphor dehydrogenase gene on the cam plasmid of pseudomonas putida (atcc 17453).	we determined the complete nucleotide sequence of the first gene of pseudomonas putida cytochrome p-450cam hydroxylase operon, camd, which encodes 5-exo-hydroxycamphor dehydrogenase. this dehydrogenase apparently consists of 361 amino acids and has a molecular mass of 38.4 kda. structural relationships to other zinc-containing alcohol dehydrogenases also became evident.	1993	8334169
a t7 rna polymerase-based system for the construction of pseudomonas strains with phenotypes dependent on tol-meta pathway effectors.	a general method to construct recombinant pseudomonas putida (and related bacteria), which transcribe specific genes inserted into their chromosome in response to the presence of alkyl- and halobenzoates, has been developed. the system is based on the ability of the t7 rna polymerase (t7pol) to initiate transcription from cognate promoter sequences located upstream from cloned genes. a specialized transposon, mini-tn5 xyls/pm::t7pol, was constructed which contains the structural t7 gene 1 downst ...	1993	8244019
the specific growth rate of pseudomonas putida paw1 influences the conjugal transfer rate of the tol plasmid.	the kinetics of the conjugal transfer of a tol plasmid were investigated by using pseudomonas putida paw1 as the donor strain and p. aeruginosa pao 1162 as the recipient strain. short-term batch mating experiments were performed in a nonselective medium, while the evolution of the different cell types was determined by selective plating techniques. the experimental data were analyzed by using a mass action model that describes plasmid transfer kinetics. this method allowed analysis of the mating ...	1993	8250565
plasmid-mediated mineralization of naphthalene, phenanthrene, and anthracene.	the well-characterized plasmid-encoded naphthalene degradation pathway in pseudomonas putida ppg7(nah7) was used to investigate the role of the nah plasmid-encoded pathway in mineralizing phenanthrene and anthracene. three pseudomonas strains, designated 5r, dfc49, and dfc50, were recovered from a polynuclear aromatic hydrocarbon-degrading inoculum developed from a manufactured gas plant soil slurry reactor. plasmids pka1, pka2, and pka3, approximately 100 kb in size, were isolated from these st ...	1993	8328809
gene components responsible for discrete substrate specificity in the metabolism of biphenyl (bph operon) and toluene (tod operon).	bph operons coding for biphenyl-polychlorinated biphenyl degradation in pseudomonas pseudoalcaligenes kf707 and pseudomonas putida kf715 and tod operons coding for toluene-benzene metabolism in p. putida f1 are very similar in gene organization as well as size and homology of the corresponding enzymes (g. j. zylstra and d. t. gibson, j. biol. chem. 264:14940-14946, 1989; k. taira, j. hirose, s. hayashida, and k. furukawa, j. biol. chem. 267:4844-4853, 1992), despite their discrete substrate rang ...	1993	8349562
[the role of pyrimidines in the biosynthesis of the fluorescing pigment pyoverdin pm in pseudomonas putida m].	dihydroorotate was shown to be a predecessor of deoxyquinoline nucleus of a fluorescing enzyme pyoverdin pm in pseudomonas putida m. the data was obtained in experiments with a set of pyr- mutants with different steps of pyrimidine synthesis blocked. a scheme for deoxyquinoline nucleus of the enzyme including dihydroorotate participation is proposed.	1993	8289842
[microbial enzymes and their inhibitors].	several proteolytic enzymes and dehydrogenases of microbial origin were studied with special regard to structure-activity relationship. enzyme genes of zn-proteases, subtilisin and pyroglutamyl aminopeptidase from genus bacillus, prolyl endopeptidases from flavobacterium and aeromonas, and of protease ii from e. coli were cloned, sequenced and overproduced in e. coli, their active site structures being elucidated by chemical modification as well as by site-directed mutagenesis. homology analysis ...	1993	8254479
reinvestigation of the role of thiol groups of glyoxalase i purified from yeast (saccharomyces cerevisiae).	glyoxalase i has been purified to homogeneity from saccharomyces cerevisiae and tested with two different thiol reagents, i.e., 5,5'-dithiobis-(2-nitrobenzoic acid) (dtnb) and 1-chloro-2,4-dinitrobenzene (cdnb). dtnb reacts with four thiol groups per molecule of enzyme and leads to a complete inhibition which is not reversed by addition of the disulfide-reducing agent dithiothreitol. on the other hand, cdnb slightly affects the glyoxalase-i activity and alkylates only one thiol residue/enzyme. i ...	1993	8373819
a dna module encoding bph genes for the degradation of polychlorinated biphenyls (pcbs).	in this report we describe the development and construction of a dna module which encodes bph genes for the metabolism of pcbs and which is capable of stable integration into the chromosome of gram negative bacteria. introduction of the bph-module into pseudomonas putida kt2442, pseudomonas sp. strain b13 and its genetically engineered derivative b13fr1 expanded the biodegradative ability of these strains to include biphenyl and 4-chlorobiphenyl. the bph operon was stably inherited under laborat ...	1993	8262365
the effects of modified atmospheres on the growth of psychrotrophic pseudomonads on a surface in a model system.	atmospheres containing concentrations of co2 as low as 20% (balance nitrogen) inhibited the growth of pseudomonas fluorescens and pseudomonas putida on the surface of buffered brain heart infusion agar plates, ph 6.8, incubated at 5 or 15 degrees c in flexible packages. the modified atmospheres decreased the growth rates and reduced the populations attained at the end of the exponential phase of growth, but had no substantial effect on the lag phase. p. fluorescens was less tolerant of co2 than ...	1993	8268059
microbial metabolism of quinoline and related compounds. xx. quinaldic acid 4-oxidoreductase from pseudomonas sp. ak-2 compared to other procaryotic molybdenum-containing hydroxylases.	quinaldic acid 4-oxidoreductase from pseudomonas sp. ak-2 catalyses the hydroxylation of quinoline 2-carboxylic (quinaldic acid) to 4-hydroxyquinoline 2-carboxylic acid (kynurenic acid) with concomitant reduction of a suitable electron acceptor. an analogous hydroxylation in para-position relative to the n-heteroatom was only recently described for quinaldine 4-oxidoreductase (de beyer & lingens, 1993, biol. chem. hoppe-seyler 374, 101-110) and for quinaldic acid 4-oxidoreductase from serratia m ...	1993	8292263
early and late responses of tol promoters to pathway inducers: identification of postexponential promoters in pseudomonas putida with lacz-tet bicistronic reporters.	transcriptional lacz fusions to the pu and pm promoters of the tol (toluene degradation) plasmid inserted in monocopy in the chromosome of pseudomonas putida showed a very different responsiveness to their respective aromatic effectors regarding growth phase. while a substantial xyls-dependent activation of pm-lacz was detected nearly instantly after m-toluate addition, xylr- and xylene-mediated induction of the sigma 54 promoter pu became significant only after cells slowed down exponential gro ...	1993	8226632
essentiality of the three carboxyl-terminal amino acids of the plasmid rk2 replication initiation protein trfa for dna binding and replication activity in gram-negative bacteria.	in a previous study of mutations in trfa, the gene encoding the replication initiation protein of the broad host-range plasmid rk2, a carboxyl-terminal deletion of 3 amino acids of the trfa protein was found to be completely nonfunctional for rk2 replication in escherichia coli and other gram-negative bacteria. in this work site-directed mutagenesis of the trfa gene was carried out to construct trfa proteins altered in the three carboxyl-terminal positions. specifically, trfa proteins with delet ...	1993	8227054
the oct plasmid encodes d-lysine membrane transport and catabolic enzymes in pseudomonas putida.	pseudomonas putida (oleovorans) (pp(oct)) cured of its oct plasmid (pp) no longer grows on d-lysine. conjugation of pptrp- with three different methionine auxotrophs carrying the oct plasmid resulted in pptrp- (oct) organisms that grew on d-lysine. three early d-lysine catabolic enzymes encoded by the oct plasmid are a lysine racemase, the proposed conversion of d-lysine to delta 1-piperidine-2-carboxylate (p2c), for which we provide evidence, and p2c reductase which converts p2c to pipecolate. ...	1993	8234494
depletion of serum methionine by methioninase in mice.	methionine dependence is a tumor-specific metabolic defect found in human cancer cell lines as well as in fresh human tumor specimens. methionine dependent tumors cease growing when deprived of methionine, unlike normal cells which can substitute homocysteine for methionine for their growth requirement. we have previously purified a stable, endotoxin-free methioninase from the bacterium, pseudomonas putida. we demonstrate in this report that purified methioninase can lower the serum levels of me ...	1993	8239522
identification of ser143 as the site of modification in the active site of histidine ammonia-lyase.	histidine ammonia-lyase (histidase) from pseudomonas putida was irreversibly inactivated by l-cysteine at ph 10.5 in the presence of oxygen. inactivation was accompanied by the formation of a new uv-absorbing species centered around 340 nm. l-[35s]cysteine labeling experiments revealed that 4 mol of l-cysteine was bound per mole of enzyme tetramer upon complete modification. however, the radiolabel was dissociated from the protein under denaturing conditions without loss of the 340-nm absorbance ...	1993	8239649
metabolism of dibenzothiophene and naphthalene in pseudomonas strains: complete dna sequence of an upper naphthalene catabolic pathway.	from a soil isolate, pseudomonas strain c18, we cloned and sequenced a 9.8-kb dna fragment that encodes dibenzothiophene-degrading enzymes. nine open reading frames were identified and designated doxabdefghij. collectively, we refer to these genes as the dox pathway. at the nucleotide level, doxabd are identical to the ndoabc genes that encode naphthalene dioxygenase of pseudomonas putida. the doxg protein is 97% identical to nahc (1,2-dihydroxynaphthalene dioxygenase) of p. putida. doxe has 37% ...	1993	8226631
a mutagenesis system utilizing a tn1722 derivative containing an escherichia coli-specific vector plasmid: application to pseudomonas species.	a novel transposon (tn) mutagenesis system for gram- non-enteric bacteria was developed which allowed rapid and one-step cloning of the mutated region in escherichia coli. the tn constructed was tn1722-299km, a tn1722 derivative containing a kmr gene and the entire sequence of an e. coli-specific plasmid, pacyc184. the hybrid plasmid consisting of tn1722-299km and the transfer genes of plasmid r388 was conjugally transferred from e. coli to pseudomonas putida or p. aeruginosa, and selection of t ...	1993	8294012
in-vivo-generated fusion promoters in pseudomonas putida.	plasmid pest1463 carrying the promoterless pheba operon was cloned into pseudomonas putida paw85, and phenol-utilizing colonies were isolated on minimal plates containing phenol as the only carbon and energy source. in these clones, chromosomally located tn4652 was transposed upstream from the coding sequencing of phea (encoding phenol monooxygenase). sequence analysis together with mapping of the transcription start point of the pheba operon in the recombinant plasmids revealed that fusions of ...	1993	8387446
growth and plasmid-encoded naphthalene catabolism of pseudomonas putida in batch culture.	the growth characteristics of pseudomonas putida plasmid-harbouring strains which catabolize naphthalene via various pathways in batch culture with naphthalene as the sole source of carbon and energy have been investigated. the strains under study were constructed using the host strain p. putida bs394 which contained various naphthalene degradation plasmids. the highest specific growth rate was ensured by the plasmids that control naphthalene catabolism through the meta-pathway of catechol oxida ...	1993	8270196
construction of chromosomal reca mutants of pseudomonas putida ppg2.	the reca gene of pseudomonas putida ppg2 was cloned by complementation of the reca mutations of escherichia coli strains dh5 alpha and hb101. the nucleotide sequence of the dna fragment was determined and shown to contain reca and a downstream partial open reading frame. two mutants of p. putida ppg2, strains js387 and js388, were constructed by insertional inactivation of reca with a tetracycline-resistance gene in both orientations. both mutants acquired sensitivity to methyl methanesulfonate ...	1993	8294013