Showing papers on "Pseudomonas putida published in 1974"
TL;DR: Preliminary results from mutant strains of Pseudomonas putida mt-2 and from a mutant defective in the regulation of the plasmid-carried pathway suggest that the wild type contains two benzoate oxidase systems, one on thePlasmid which is nonspecific in both its catalysis and its induction and one in the chromosome which is more specific to benzoates as substrate and is specifically induced by benzosate.
Abstract: Mutant strains of Pseudomonas putida (arvilla) mt-2 which have lost the ability to grow at the expense of m- or p-toluate (methylbenzoate) but retain the ability to grow with benzoate arise spontaneously during growth on benzoate; this genetic loss occurs to a lesser extent during growth on nonaromatic carbon sources in the presence of mitomycin C. The mutants have totally lost the activity of the enzymes of the divergent meta pathway with the possible exception of 2-oxopent-4-enoate hydratase and 4-hydroxy-2-oxovalerate aldolase; unlike the wild type they utilize benzoate by the ortho pathway. Evidence is presented that these mutants have lost a plasmid coding for the enzymes of the meta pathway, which may be transmitted back to them or into other P. putida strains. Preliminary results from these mutants and from a mutant defective in the regulation of the plasmid-carried pathway suggest that the wild type contains two benzoate oxidase systems, one on the plasmid which is nonspecific in both its catalysis and its induction and one on the chromosome which is more specific to benzoate as substrate and is specifically induced by benzoate.
483 citations
TL;DR: Two new metabolites, 5-carboxymethyl-2-hydroxymuconic acid and 2-hydroxyhepta-2,4-diene-1,7-dioic acid, were isolated from reaction mixtures and a third, 4-Hydroxyphenylacetic acid, was shown to be cleaved by extracts to give pyruvate and succinic semialdehyde.
Abstract: A species of Acinetobacter and two strains of Pseudomonas putida when grown with 4-hydroxyphenylacetic acid gave cell extracts that converted 3,4-dihydroxyphenylacetic acid (homoprotocatechuic acid) into carbon dioxide, pyruvate, and succinate. The sequence of enzyme-catalyzed steps was as follows: ring-fission by a 2,3-dioxygenase, nicotinamide adenine dinucleotide-dependent dehydrogenation, decarboxylation, hydration, aldol fission, and oxidation of succinic semialdehyde. Two new metabolites, 5-carboxymethyl-2-hydroxymuconic acid and 2-hydroxyhepta-2,4-diene-1,7-dioic acid, were isolated from reaction mixtures and a third, 4-hydroxy-2-ketopimelic acid, was shown to be cleaved by extracts to give pyruvate and succinic semialdehyde. Enzymes of this metabolic pathway were present in Acinetobacter grown with 4-hydroxyphenylacetic acid but were effectively absent when 3-hydroxyphenylacetic acid or phenylacetic acid served as sources of carbon.
130 citations
TL;DR: Cells of Pseudomonas putida, after growth with naphthalene as sole source of carbon and energy, contain an enzyme that oxidizes (+)-cis-1(r),2(s)-dihydroxy-1,2- dihydronaphthalenes to 1,2
Abstract: Cells of Pseudomonas putida, after growth with naphthalene as sole source of carbon and energy, contain an enzyme that oxidizes (+)-cis-1(r),2(s)-dihydroxy-1,2-dihydronaphthalene to 1,2-dihydroxynaphthalene. The purified enzyme has a molecular weight of 102,000 and apparently consists of four 25,500 molecular weight subunits. The enzyme is specific for nicotinamide adenine dinucleotide as an electron acceptor and also oxidizes several other cis-dihydrodiols. However, no enzymatic activity was observed with trans-1,2-dihydronaphthalene, or the K-region cis-dihydrodiols of carcinogenic polycyclic hydrocarbons.
116 citations
TL;DR: Pseudomonas putida 39/D oxidized m-xylene to a compound whose spectral and chromatographic characteristics were consistent with the structure 3, 5-dimethyl-3,5-cyclohexadiene-1,2-diol, which was very unstable, and all attempts to isolate it led to the formation of 2,4-Dimethylphenol.
Abstract: Pseudomonas putida 39/D oxidized p-xylene to cis-3,6-dimethyl-3,5-cyclohexadiene-1,2-diol (cis-p-xylene dihydrodiol). The latter compound was isolated in crystalline form and its physical properties were determined. The cis configuration of the hydroxyl groups in the oxidation product was inferred from its ability to form an isopropylidene derivative with 2,2-dimethoxypropane. Acid treatment of cis-p-xylene dihydrodiol resulted in the formation of 2,5-dimethylphenol. A partially purified preparation of cis-toluene dihydrodiol dehydrogenase oxidized cis-p-xylene dihydrodiol to 1,2-dihydroxy-3,6-dimethylbenzene (3,6-dimethylpyrocatechol). P. putida 39/D oxidized m-xylene to a compound whose spectral and chromatographic characteristics were consistent with the structure 3,5-dimethyl-3,5-cyclohexadiene-1,2-diol. This product was very unstable, and all attempts to isolate it led to the formation of 2,4-dimethylphenol.
102 citations
TL;DR: The microbial cells of Pseudomonas putida ATCC 4359 were immobilized by entrapment in a polyacrylamide gel lattice and the permeability of substrate or product through the cell wall und the heat stability of the enzyme were increased by immobilization of the cells.
Abstract: The microbial cells of Pseudomonas putida ATCC 4359 were immobilized by entrapment in a polyacrylamide gel lattice. Enzymatic properties of L‐arginine deiminase of the immobilized P. putida cells were investigated and compared with those of the intact cells. The permeability of substrate or product through the cell wall und the heat stability of the enzyme were increased by immobilization of the cells. No difference was observed between pH activity curves of the intact and immobilized cells. The optimal temperature for the formation of L‐citrulline was 37°C for the intact cells and 55° C for the immobilized cells.
98 citations
TL;DR: Evidence was obtained that the dehydrogenase catalyzed the oxidation of 2- methyl-3-hydroxybutyryl-CoA to 2-methylacetoacetyl- CoA and that these two enzymes appeared to be different dehydrogenases.
Abstract: Pseudomonas putida oxidized isoleucine to acetyl-coenzyme A (CoA) and propionyl-CoA by a pathway which involved deamination of d-isoleucine by oxidation and l-isoleucine by transamination, oxidative decarboxylation, and beta oxidation at the ethyl side chain. At least three separate inductive events were required to form all of the enzymes of the pathway: d-amino acid dehydrogenase was induced during growth in the presence of d-isoleucine; branched-chain keto dehydrogenase was induced during growth on 2-keto-3-methylvalerate and enzymes specific for isoleucine metabolism; tiglyl-CoA hydrase and 2-methyl-3-hydroxybutyryl-CoA dehydrogenase were induced by growth on isoleucine, 2-keto-3-methylvalerate, 2-methylbutyrate, or tiglate. Tiglyl-CoA hydrase and 2-methyl-3-hydroxybutyryl-CoA dehydrogenase were purified simultaneously by several enzyme concentration procedures, but were separated by isoelectric focusing. Isoelectric points, pH optima, substrate specificity, and requirements for enzyme action were determined for both enzymes. Evidence was obtained that the dehydrogenase catalyzed the oxidation of 2-methyl-3-hydroxybutyryl-CoA to 2-methylacetoacetyl-CoA. 2-Methyl-3-hydroxybutyryl-CoA dehydrogenase catalyzed the oxidation of 3-hydroxybutyryl-CoA, but l-3-hydroxyacyl-CoA dehydrogenase from pig heart did not catalyze the oxidation of 2-methyl-3-hydroxybutyryl-CoA; therefore, they appeared to be different dehydrogenases. Furthermore, growth on tiglate resulted in the induction of tiglyl-CoA hydrase and 2-methyl-3-hydroxybutyryl-CoA dehydrogenase, but these two enzymes were not induced during growth on crotonate or 3-hydroxybutyrate.
57 citations
TL;DR: This finding implies that lysine racemase in this strain, while detectable in cell extracts, is not physiologically functional in intact cells at a rate that would permit growth of mutants blocked in the l-lysine pathway, suggesting a mechanism in bacteria for maintaining regulatory patterns in pathways that may have lost their capacity to support growth.
Abstract: The isolation of several mutant strains blocked in l-lysine degradation has permitted an assessment of the physiological significance of enzymatic reactions related to lysine metabolism in Pseudomonas putida. Additional studies with intact cells involved labeling of metabolic intermediates from radioactive l- or d-lysine, and patterns of enzyme induction in both wild-type and mutant strains. These studies lead to the conclusions that from l-lysine, the obligatory pathway is via delta-aminovaleramide, delta-aminovalerate, glutaric semialdehyde, and glutarate, and that no alternative pathways from l-lysine exist in our strain. A distinct pathway from d-lysine proceeds via Delta(1)-piperideine-2-carboxylate, l-pipecolate, and Delta(1)-piperideine-6-carboxylate (alpha-aminoadipic semialdehyde). The two pathways are independent in the sense that certain mutants, unable to grow on l-lysine, grow at wild-type rates of d-lysine, utilizing the same intermediates as the wild type, as inferred from labeling studies. This finding implies that lysine racemase in our strain, while detectable in cell extracts, is not physiologically functional in intact cells at a rate that would permit growth of mutants blocked in the l-lysine pathway. Pipecolate oxidase, a d-lysine-related enzyme, is induced by d-lysine and less efficiently by l-lysine. Aminooxyacetate virtually abolishes the inducing activity of l-lysine for this enzyme, suggesting that lysine racemase, although functionally inactive for growth purposes, may still have regulatory significance in permitting cross-induction of d-lysine-related enzymes by l-lysine, and vice versa. This finding suggests a mechanism in bacteria for maintaining regulatory patterns in pathways that may have lost their capacity to support growth. In addition, enzymatic studies are reported which implicate Delta(1)-piperideine-2-carboxylate reductase as an early step in the d-lysine pathway.
56 citations
TL;DR: Highly purified P- 450 cam and P-450 LM show immunological cross reaction by competitive binding and inhibition of catalytic activity and are of similar subunit molecular weight and amino acid composition.
51 citations
TL;DR: Pseudomonas putida NCIB 10015 metabolizes phenol and the cresols (methylphenols) by the meta pathway and metabolizes benzoate by the ortho pathway and the role of catechol andThe methylcatechols as inducers is discussed.
Abstract: Pseudomonas putida NCIB 10015 metabolizes phenol and the cresols (methylphenols) by the meta pathway and metabolizes benzoate by the ortho pathway. Growth on catechol, an intermediate in the metabolism of both phenol and benzoate, induces both ortho and meta pathways; growth on 3- or 4-methylcatechols, intermediates in the metabolism of the cresols, induces only the meta pathway to a very limited degree. Addition of catechol at a growth-limiting rate induces virtually no meta pathway enzymes, but high levels of ortho pathway enzymes. The role of catechol and the methylcatechols as inducers is discussed. A method is described for assaying low levels of catechol 1,2-oxygenase in the presence of high levels of catechol 2,3-oxygenase and vice versa.
50 citations
TL;DR: The iron-sulfur protein putidaredoxin from Pseudomonas putida contains 106 amino acids arranged in a single peptide chain and it is suggested that positions 39, 45, 48, and 85 (or 86) may be chelated to the iron in the iron sulfide reactive center.
48 citations
TL;DR: The structure of 2-hydroxy-6-oxo- 6-phenylhexa-2,4-dienoic acid for the meta-cleavage product of 2,3-dihydroxybiphenyl by a Pseudomonas putida strain was demonstrated on the basis of its chemical and physicochemical properties and those of its derivatives.
Abstract: The structure of 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoic acid for the meta -cleavage product of 2,3-dihydroxybiphenyl by a Pseudomonas putida strain was demonstrated on the basis of its chemical and physicochemical properties and those of its derivatives.
TL;DR: A selection procedure is described which was used to isolate mutants of Pseudomonas putida strain U in the following enzymes of the meta-fission pathway of phenol and cresols: hydrolase, tautomerase, and decarboxylase.
Abstract: A selection procedure is described which was used to isolate mutants of Pseudomonas putida strain U in the following enzymes of the meta-fission pathway of phenol and cresols: hydrolase, tautomerase, and decarboxylase Strains deficient in the hydrolase are unable to use either o- or m-cresol as a sole carbon source and were shown to accumulate 2-hydroxy-6-keto-2,4-heptadienoate when incubated in the presence of o- or m-cresol When 2-hydroxymuconic semialdehyde (plus nicotinamide adenine dinucleotide, oxidized form) was metabolized by phenol-induced extracts of tautomerase-deficient strains, the enol tautomer of 4-oxalocrotonate accumulated and was then converted slowly to the keto tautomer by a nonenzymatic reaction Phenol-induced extracts of decarboxylase-deficient strains accumulated the keto tautomer of 4-oxalocrotonate from 2-hydroxymuconic semialdehyde (plus nicotinamide adenine dinucleotide, oxidized form) Strains with an inactive decarboxylase are unable to completely metabolize either phenol or p-cresol Tautomerase-defective strains are unable to grow with p-cresol as the sole carbon source and grow only very slowly on phenol
TL;DR: The infectious plasmid OCT, which specifies a set of dissimilatory enzymes responsible for the degradation of n-octane, has been shown to be an aggregate of a noninfectious OCT plasmide and an infectious plasid with sex factor activity, which appears to be compatible with another degradative plasmids, CAM.
Abstract: The infectious plasmid OCT, which specifies a set of dissimilatory enzymes responsible for the degradation of n-octane, has been shown to be an aggregate of a noninfectious OCT plasmid and an infectious plasmid with sex factor activity. The infectious plasmid, which can be eliminated from the cells of Pseudomonas putida by mitomycin C treatment without loss of the OCT plasmid and vice versa, has been designated as factor K. The infectious plasmid (factor K) is not only responsible for the mobilization of OCT, but also mobilizes chromosomal genes at a frequency of 10−2 to 10−3 per donor cell. Whereas OCT is incompatible with another degradative plasmid, CAM, factor K appears to be compatible with it.
TL;DR: Kinetic parameters of uptake and transport and studies on succinate metabolism provide evidence consistent with concurrent increases in transport and metabolism to account for induced succinate uptake by P. putida.
TL;DR: Detailed electrophoretic, structural, immunological, and genetic studies show Pseudomonas putida's two induced catabolic pathways to be distinct proteins, although closely similar in most kinetic properties studied, as well as in molecular weight, subunit status, and amino acid composition.
TL;DR: Findings confirm the hypothesis that this iron-sulfur protein is the terminal oxidase within this enzyme system, isolated from P. puti& grown on 4methoxybenzoate as the sole carbon source and separated into two components.
TL;DR: The apparently close evolutionary homology of the muconolactone isomerases and the wide evolutionary separation of Acinetobacter and Pseudomonas suggest that the structural gene for the catabolic enzyme may have been exchanged well after the divergence of other genes that dictated the evolutionary separated of the two genera.
TL;DR: The transfer of the OCT plasmid from Pseudomonas oleovorans to Pseudomanas putida strain PpGl results in the acquisition of three independent replicons: OCT, factor K, and the MER plasmids.
Abstract: The transfer of the OCT plasmid from Pseudomonas oleovorans to Pseudomonas putida strain PpGl results in the acquisition of three independent replicons: OCT, factor K, and the MER plasmid. OCT is a nontransmissible plasmid harboring genes that code for the enzymes responsible for the degradation of n-octane. Factor K is a transfer plasmid capable of mobilizing OCT as well as chromosomal genes but incapable of enhancing transfer frequencies of other transmissible plasmids such as CAM, SAL, or RP-1. MER is a self-transmissible plasmid which can confer resistance to high concentrations of mercury salts. While OCT and MER are incompatible with CAM, factor K is compatible with it. Transmissible plasmids such as SAL, CAM, MER, or RP-1 cannot mobilize OCT to any significant extent, and exert strong repression on factor K-mediated transfer of chromosomal genes.
TL;DR: A mutant strain of Pseudomonas putida NCIB 10015 has been isolated that is unable to utilize para -cresol as a sole carbon source and preliminary evidence is presented that this mutant strain has a regulatory element of altered specificity.
TL;DR: The sequence of the 46 NH2-terminal residues of the tryptophan synthetase α chain of Bacillus subtilis was determined and compared with the corresponding sequences of Escherichia coli, Shigella dysenteriae, Salmonella typhimurium, Aerobacter aerogenes, Serratia marcescens, and Pseudomonas putida.
Abstract: The sequence of the 46 NH2-terminal residues of the tryptophan synthetase α chain of Bacillus subtilis was determined and compared with the corresponding sequences of Escherichia coli, Shigella dysenteriae, Salmonella typhimurium, Aerobacter aerogenes, Serratia marcescens, and Pseudomonas putida. A deletion of six residues was found at the NH2-terminal end of the α chain of B. subtilis.
TL;DR: The efficiency of substrate entry and catalysis near 0 C are factors in the ability of this facultative psychrophile to grow in a histidine medium at 0 C.
Abstract: Pseudomonas putida was able to grow at 0 C in a complex medium containing l-histidine and to synthesize histidine ammonia-lyase and urocanase. The activity of the former enzyme was assessed between −10 and 60 C in cells and in cell extracts. Activity was maximal from 20 to 35 C. Below 20 C, activity decreased with temperature but, significantly, the enzyme exhibited 30% of its maximal activity at 1.5 C. The temperature response was similar in both intact cells and cell extracts, which indicated that the cell membrane did not significantly limit the entry of histidine at low temperature. Above and below the maximal temperature range, the reduced activity was not caused by irreversible inactivation, as shown by preincubation experiments. Also, when the temperature was rapidly changed from 60 to 30 C during an assay, the reaction rate increased abruptly to the full 30 C activity without a lag. This demonstrated the rapid reversibility of inactivation. The apparent Michaelis constant increased with temperature. As the substrate concentration was decreased, the enzyme activity became less dependent on temperature. The efficiency of substrate entry and catalysis near 0 C are factors in the ability of this facultative psychrophile to grow in a histidine medium at 0 C.
Journal Article•
TL;DR: A rapid identification method of glucose nonfermentative gram-negative rods was established and 320 strains isolated were divided into five groups according to their characteristics in pigmentation, acid from glucose, cytochrome oxidase activity and motility.
Abstract: A rapid identification method of glucose nonfermentative gram-negative rods was established and 320 strains isolated were divided into five groups according to their characteristics in pigmentation, acid from glucose, cytochrome oxidase activity and motility Further characterization of the strains in each group resulted in the identification that the strains in group I were Pseudomonas aeruginosa , strains in group II, P aeruginosa and Pseudomonas putida Achromogenic strains of P aeruginosa were classified into group III, Pseudomonas maltophilia, Pseudomonas alcaligenes and Alcaligenes faecalis into group IV and Acinetobacter calcoaceticus (Acinetobacter anitratus and Achromobacter lwoffii) in group V When fluorescent pigment production was taken as a standard, 259 out of 263 chromogenic strains were identified as P aeruginosa and the remaining four were P putida Whereas forty-five achromogenic strains included twenty-four A calcoaceticus, eight P aeruginosa, six A faecalis, five Pmaltophilia and two P alcaligenes From May 1970 to June 1971, 368 strains of glucose nonfermentative rods were isolated from clinical specimens sent to the Central Laboratories of Tohoku University Hospital and three fourth (286/368) of the isolates were P aeruginosa
TL;DR: It is proposed to term these naturally occurring plasmids “degradative plasmid” because of the selective nutritional advantage they offer to the host cells harboring them.
Abstract: Considerable attention has recently been directed toward studying the genetic basis of nutritional versatility in Pseudomonas. Unlike the enterobacteria, which grow on a limited number of complex organic compounds, the genus Pseudomonas is known for its ability to derive its carbon and energy from the dissimilation of a large number of organic compounds of varying complexities (1). The genetic and physiological regulation, the enzymology, and the biochemistry of a large number of catabolic pathways have therefore been extensively studied in this genus (2). The ability of the pseudomonads to degrade a number of complex organic compounds has been recently shown to be due to their possession of extrachromosomal elements that specify the entire enzyme complements of a particular degradative pathway. Thus the genes specifying enzymes responsible for the degradation of salicylate in a strain of Pseudomonas putida R1 are not part of the bacterial chromosome but are borne on extrachromosomal elements which are transmissible among a number of Pseudomonas species (3). Similarly, the camphor degradative pathway, comprised of a set of 15-20 inducible enzymes, is specified by genes that are also part of a transmissible plasmid (4). Another set of genes that code for enzymes responsible for the metabolism of octane is carried on yet another transmissible plasmid (5). We have proposed to term these naturally occurring plasmids “degradative plasmids” because of the selective nutritional advantage they offer to the host cells harboring them (6).
01 Jan 1974
TL;DR: Pseudomonas putida NCIB10015 metabolizes phenol and thecresols (methylphenols) by the metapathway and benzoate by theortho pathway and the role of catechol and themethylcatechols asinducers is discussed.
Abstract: Pseudomonas putida NCIB10015metabolizes phenol andthecresols (methylphenols) bythemetapathway andmetabolizes benzoate bytheortho pathway. Growth on catechol, an intermediate inthemetabolism ofbothphenol and benzoate, induces bothortho andmetapathways; growth on 3-or4-methylcatechols, intermediates inthemetabolism ofthecresols, induces onlythemeta pathway toa verylimited degree. Addition ofcatechol atagrowth-limiting rate induces virtually no metapathway enzymes, buthighlevels ofortho pathway enzymes.Therole ofcatechol andthemethylcatechols asinducers isdiscussed. A methodisdescribed forassaying lowlevels ofcatechol 1,2-oxygenase inthe presenceofhighlevels ofcatechol 2,3-oxygenase andvice versa.