Publications
| Title | Abstract | Year(sorted ascending) Filter | PMID Filter |
|---|
| inactivation of delta 5-3-oxo steroid isomerase with active-site-directed acetylenic steroids. | several steroid analogues containing conjugated acetylenic ketone groups as part of a seco-ring structure or as substituents on the intact steroid system are irreversible inhibitors of delta 5-3-oxo steroid isomerase (ec 5.3.3.1) from pseudomonas testosteroni. thus 10 beta-(1-oxoprop-2-ynyl)oestr-4-ene-3,17-dione (i), 5,10-seco-oestr-4-yne-3,10,17-trione (ii), 17 beta-hydroxy-5,10-seco-oestr-4-yne-3,10-dione (iii) and 17 beta-(1-oxoprop-2-ynyl)androst-4-en-3-one (iv) irreversibly inactivate isom ... | 1981 | 7305923 |
| induction of 3-hydroxybenzoate 2-hydroxylase in a pseudomonas testosteroni mutant. | benzoate was established as the inducer of a unique 3-hydroxybenzoate 2-hydroxylase activity found in a pseudomonas testosteroni mutant which is unable to grow on m-hydroxybenzoate as its sole source of carbon and energy. | 1982 | 7054148 |
| testosterone-dependent oxygen consumption in membrane vesicles of pseudomonas testosteroni. | oxygen consumption was measured in membrane vesicles of pseudomonas testosteroni using conditions similar to those identified for testosterone transport in these vesicles. testosterone and nad+, which are primary requirements for testosterone transport, were both required for maximum oxygen consumption suggesting that testosterone transport and oxygen consumption were linked. testosterone-dependent oxygen consumption was inhibited by 95% by 1 mm kcn indicating that the electron-transport chain c ... | 1982 | 7109594 |
| mass spectrometric studies of a modified active-site tetrapeptide from delta 5-3-ketosteroid isomerase of pseudomonas testosteroni. | examination of the product of affinity labeling of delta 5-3-ketosteroid isomerase (ec 5.3.3.1) of pseudomonas testosteroni by the suicide substrate [7-3h]5,10-secoestr-5-yne-3,10,17-trione has demonstrated that the steroid becomes bound by an acid- and base-labile linkage to an active-site peptide representing residues 55-58 (h2n-tyr-ala-asn-ser-co2h) of the primary structure (penning, t. m., and talalay, p. (1981) j. biol. chem. 256, 6851-6858). upon release of the steroid by mild acid hydroly ... | 1982 | 7130168 |
| proton nuclear magnetic resonance studies of pseudomonas testosteroni 3-oxo-delta 5-steroid isomerase and its interaction with 17 beta-estradiol. | 1982 | 6295444 | |
| 2-pyrone-4,6-dicarboxylic acid, a catabolite of gallic acids in pseudomonas species. | 2-pyrone-4,6-dicarboxylate hydrolase was purified from 4-hydroxybenzoate-grown pseudomonas testosteroni. gel filtration and electrophoretic measurements indicated that the preparation was homogeneous and gave a molecular weight of 37,200 for the single subunit of the enzyme. hydrolytic activity was dependent upon a functioning sulfhydryl group(s) and was freely reversible; the equilibrium position was dependent upon ph, with equimolar amounts of pyrone and open-chain form present at ph 7.9. sinc ... | 1982 | 7142106 |
| partial purification and characterization of a membrane-associated steroid-binding protein from pseudomonas testosteroni. | a steroid-binding protein obtained from the supernatant of the final wash from the preparation of membrane vesicles was purified severalfold to near homogeneity. the protein binds c18 and c19 steroids but has the highest affinity for androstenedione (kd = 1.6 x 10(-10) m). the molecular weight is 51,000 - 58,000. binding activity is slightly inhibited by cu2+, ca2+, and mg2+ and completely inhibited by zn2+. the protein has no detectable steroid degradative activity. analysis of androstenedione ... | 1982 | 6889907 |
| photoaffinity modification of delta 5-3-ketosteroid isomerase by light-activatable steroid ketones covalently coupled to agarose beads. | in order to identify the minor site(s) of photoattachment of unsaturated steroid ketones to delta 5-3-ketosteroid isomerase from pseudomonas testosteroni, we have developed a solid-state photoaffinity labeling technique. two solid-state reagents, o-carboxymethylagarose-ethylenediamine-succinyl-17 beta-o-19-nortestosterone and o-carboxymethylagarose-ethylenediamine-succinyl-17 beta-o-4,6-androstadien-3-one, have been synthesized. under anaerobic conditions, isomerase bound to these resins is phot ... | 1983 | 6860646 |
| epr and mössbauer studies of protocatechuate 4,5-dioxygenase. characterization of a new fe2+ environment. | protocatechuate 4,5-dioxygenase from pseudomonas testosteroni has been purified to homogeneity and crystallized. the iron containing, extradiol dioxygenase is shown to be composed of two subunit types (alpha, mr = 17,700 and beta, mr = 33,800) in a 1:1 ratio; such a composition has not been observed for other extradiol dioxygenases. the 4.2 k mössbauer spectrum of native protocatechuate 4,5-dioxygenase prepared from cells grown in 57fe-enriched media consists of a doublet with quadrupole splitti ... | 1983 | 6317682 |
| differences in inhibition by various steroids of rat testis and pseudomonas testosteroni delta 5-3 beta-hydroxysteroid dehydrogenase. | the influence of various estrogens, progestogens and of cyanoketone (2 alpha-cyano-4,4,17 alpha-trimethylandrost-5-en-17 beta-ol-3-one) on the enzyme activity of delta 5-3 beta-hydroxysteroid dehydrogenase (hsd) (ec 1.1.1.145) was studied. extracts of pseudomonas testosteroni, rat testis total homogenate and a microsomal preparation were used as enzyme sources. spectrophotometric determinations and the conversion of 3h-labelled dehydroepiandrosterone to androstenedione were used to assay for enz ... | 1983 | 6603965 |
| the effect of 2,4-dinitrophenol on steroid transport in membrane vesicles of pseudomonas testosteroni. | steroid transport in pseudomonas testosteroni membrane vesicles was significantly inhibited by the uncoupled 2,4-dinitrophenol (dnp). inhibition of steroid transport was not due to inhibition of the 3 beta- and 17 beta-hydroxysteroid dehydrogenase by concentrations of up to 1 mm dnp. however, inhibition of this membrane-bound enzyme was measured at 10 mm dnp. the solubilized 3 beta- and 17 beta-hydroxysteroid dehydrogenase was more sensitive, being inhibited at both 1 and 10 mm dnp indicating a ... | 1983 | 6827840 |
| immunological relatedness of histidine ammonia-lyases from some species of pseudomonas: taxonomic implication. | 1. histidine ammonia-lyases (histidase ec 4.3.1.3) from pseudomonas testosteroni ncib 10808 and pseudomonas putida ncib 10807 were purified and specific antibody was raised to each separately in a rabbit. 2. immunological cross-reactions of each antibody to histidine ammonia-lyases from various species of pseudomonas were examined by the enzyme inhibition test. 3. the immunological data obtained suggest that these pseudomonas species can be classified into three groups. these cross-reactions ten ... | 1983 | 6862094 |
| the effect of carbonyl cyanide m-chlorophenylhydrazone on steroid transport in membrane vesicles of pseudomonas testosteroni. | the uncoupler carbonyl cyanide chlorophenylhydrazone (cccp) was an effective inhibitor of steroid transport in membrane vesicles of pseudomonas testosteroni between 10 microm and 1 microm cccp. at these concentrations the inhibition of steroid transport was not due to an inhibition of the 3 beta and 17 beta-hydroxysteroid dehydrogenase enzyme. cccp also affected testosterone-dependent oxygen consumption at concentrations up to 100 microm and inhibited respiration at 0.5 and 1 microm. the effect ... | 1983 | 6310264 |
| [transformation of escherichia coli and different species of pseudomonas by a plasmid dna isolated from pseudomonas testosteroni]. | pseudomonas testosteroni uses testosterone as sole source of carbon. we were able to isolate an extrachromosomal dna from a strain of pseudomonas testosteroni and to obtain pseudomonas putida and aeruginosa and escherichia coli transformants catabolizing testosterone. | 1983 | 6416627 |
| purification and characterization of a membrane-associated testosterone-binding protein from pseudomonas testosteroni. | a steroid-binding protein, identified in the supernatant generated when membrane vesicles of pseudomonas testosteroni are produced and harvested by centrifugation, has been purified 49-fold to homogeneity. it has a molecular weight of 30 000-35 000 and it specifically binds the c19 steroids dihydrotestosterone, testosterone, and androstenedione. it is a basic protein with an isoelectric point at ph 7.3. binding of testosterone exhibited normal saturation kinetics with an affinity constant, kd, o ... | 1983 | 6683990 |
| bacterial decarboxylation of o-phthalic acids. | the decarboxylation of phthalic acids was studied with bacillus sp. strain fo, a marine mixed culture on-7, and pseudomonas testosteroni. the mixed culture on-7, when grown anaerobically on phthalate but incubated aerobically with chloramphenicol, quantitatively converted phthalic acid to benzoic acid. substituted phthalic acids were also decarboxylated: 4,5-dihydroxyphthalic acid to protocatechuic acid; 4-hydroxyphthalic and 4-chlorophthalic acids to 3-hydroxybenzoic and 3-chlorobenzoic acids, ... | 1983 | 16346440 |
| enzymatic function in crystals of delta 5-3-ketosteroid isomerase. catalytic activity and binding of competitive inhibitors. | crystals of the steroid-metabolizing enzyme, delta 5-3-ketosteroid isomerase (ec 5.3.3.1) from pseudomonas testosteroni, exhibit many enzymatic properties. each enzyme subunit in the lattice binds a competitive inhibitor, progesterone, with the same stoichiometry (1:1) and affinity (kd = 6 x 10(-6) m) as the enzyme in solution. another competitive inhibitor, 19-nortestosterone, competes with progesterone for the same binding sites in the crystal. the enzyme crystals catalyze the conversion of de ... | 1984 | 6746640 |
| evaluation of the rapid nft system for identification of gram-negative, nonfermenting rods. | this study evaluated the ability of the rapid nft system (api system sa, montalieu-vercieu, france) to accurately identify 262 clinically isolated, gram-negative, nonfermentative rods without additional tests. identifications were classified as correct; low discrimination, with a spectrum of two or more possibilities (additional tests necessary for accurate identification); and incorrect. correct identification rates were analyzed in two categories: (i) correct to species or biotype for all orga ... | 1984 | 6490857 |
| norethisterone and ethinylestradiol do not inhibit delta 5-3 beta-hydroxysteroid dehydrogenase in rat leydig cells. | the effect of norethisterone, norethisterone acetate, ethinylestradiol and of a cyanoketone on the activity of hsd have been studied. rat leydig cells and extracts of pseudomonas testosteroni were used as enzyme sources. conversion of [3h]dhea to a and [3h]pregnenolone to progesterone were used for the assay of enzyme activity. km values for each substrate for leydig cells and for bacterial enzyme were: 1.5 x 10(-5) mol l-1 and 6.0 x 10(-5) mol l-1 dhea; 1.3 x 10(-5) mol and 7.7 x 10(-5) mol l-1 ... | 1984 | 6237898 |
| irreversible inhibition of delta 5-3-oxosteroid isomerase by 2-substituted progesterones. | 2 alpha-cyanoprogesterone (i) and 2-hydroxymethyleneprogesterone (ii) were synthesized and screened as irreversible active-site-directed inhibitors of the delta 5-3-oxosteroid isomerase (ec 5.3.3.1) from pseudomonas testosteroni. both compounds were found to inhibit the purified bacterial enzyme in a time-dependent manner. in either case the inactivated enzyme could be dialysed without return of activity, indicating that a stable covalent bond had formed between the inhibitor and the enzyme. ina ... | 1985 | 3838891 |
| mitochondrial origins. | the 16s ribosomal rna sequences from agrobacterium tumefaciens and pseudomonas testosteroni have been determined to further delimit the origin of the endosymbiont that gave rise to the mitochondrion. these two prokaryotes represent the alpha and beta subdivisions, respectively, of the so-called purple bacteria. the endosymbiont that gave rise to the mitochondrion belonged to the alpha subdivision, a group that also contains the rhizobacteria, the agrobacteria, and the rickettsias--all prokaryote ... | 1985 | 3892535 |
| mechanism of inactivation of 3-oxosteroid delta 5-isomerase by 17 beta-oxiranes. | the affinity label (17s)-spiro[estra-1,3,5(10),6,8-pentaene-17,2'-oxiran]-3-ol (5 beta) inactivates 3-oxosteroid delta 5-isomerase from pseudomonas testosteroni by formation of a covalent bond between asp-38 of the enzyme and the steroid. high-performance liquid chromatography (hplc) analysis of tryptic digests of inactivated enzyme shows that two isomeric steroid-containing peptides are formed in a ratio of 9:1 at ph 7 (tps1 and tps2). hydrolysis of each of these peptides produces a different s ... | 1985 | 4027215 |
| extraction of a steroid transport system from pseudomonas testosteroni membranes and incorporation into synthetic liposomes. | a steroid binding protein has been extracted from pseudomonas testosteroni membranes with an organic solvent system. this protection binds some c19 and c21 steroids but not c18 steroids. when this protein is incorporated into synthetic lipid vesicles constructed from p. testosteroni phospholipids, the vesicles perform concentrative uptake of testosterone in the presence of the ionophore valinomycin. this steroid binding protein is thus believed to be the steroid permease of this organism. | 1985 | 4068713 |
| use of a solid-phase photoaffinity reagent to label a steroid binding site: application to the delta 5-3-ketosteroid isomerase of pseudomonas testosteroni. | in order to extend our analysis of the reactions that occur during the active site directed photoinactivation of delta 5-3-ketosteroid isomerase sensitized by unsaturated steroid ketone photoaffinity reagents, the site of covalent attachment has been identified. a solid-phase photoaffinity reagent, delta 6-testosterone-agarose, has been employed for this purpose; this type of reagent, in contrast to solution-phase reagents, facilitated the recovery of a peptide fragment of the isomerase bearing ... | 1985 | 4092021 |
| [17o]water and nitric oxide binding by protocatechuate 4,5-dioxygenase and catechol 2,3-dioxygenase. evidence for binding of exogenous ligands to the active site fe2+ of extradiol dioxygenases. | pseudomonas testosteroni protocatechuate 4,5-dioxygenase and pseudomonas putida catechol 2,3-dioxygenase (metapyrocatechase) catalyze extradiol-type oxygenolytic cleavage of the aromatic ring of their substrates. the essential active site fe2+ of each enzyme binds nitric oxide (no) to produce an epr active complex with an electronic spin of s = 3/2. hyperfine broadening of the epr resonances of the nitrosyl complexes by 17o-enriched h2o shows that water is bound directly to the fe2+ in the nativ ... | 1985 | 2997190 |
| 3 beta, 17 beta-hydroxysteroid dehydrogenase of pseudomonas testosteroni. kinetic evidence for the bifunctional activity at a common catalytic site. | 3 beta, 17 beta-hydroxysteroid dehydrogenase (3 beta 17 beta hsdh) is an nad-dependent dehydrogenase which has a double specificity for the 3- and 17-positions on the steroid skeleton. when dehydroepiandrosterone (dhea) is used as steroid substrate, and the assay coupled with ketosteroid-isomerase, the two reactions occur alternately and each reaction on the 3-position produces a chromophoric molecule. these two reactions can follow one another without dissociation of the coenzyme from the enzym ... | 1985 | 2991019 |
| binding of 17o-labeled substrate and inhibitors to protocatechuate 4,5-dioxygenase-nitrosyl complex. evidence for direct substrate binding to the active site fe2+ of extradiol dioxygenases. | pseudomonas testosteroni protocatechuate 4,5-dioxygenase catalyzes extradiol-type oxygenolytic cleavage of the aromatic ring of its substrate. the essential active site fe2+ binds nitric oxide (no) to produce an epr active complex with an electronic spin of s = 3/2. hyperfine broadening of the epr resonances of the nitrosyl complex of the enzyme by protocatechuate (3,4-(oh)2-benzoate, pca) enriched specifically with 17o (i = 5/2) in either the 3 or the 4 hydroxyl group shows that both groups can ... | 1986 | 3003098 |
| the amino acid sequence of a delta 5-3-oxosteroid isomerase from pseudomonas putida biotype b. | we have determined the primary structure of a delta 5-3-oxosteroid isomerase from pseudomonas putida biotype b. the enzyme is a dimeric protein of two identical subunits, each consisting of a polypeptide chain of 131 residues and a mr = 14,536. the intact s-carboxymethyl protein was sequenced from the nh2 terminus using standard automated edman degradation and automated edman degradation using fluorescamine treatment at known prolines to suppress background. the isomerase was fragmented using cn ... | 1986 | 3700400 |
| quinohaemoprotein alcohol dehydrogenase apoenzyme from pseudomonas testosteroni. | cell-free extracts of pseudomonas testosteroni, grown on alcohols, contain quinoprotein alcohol dehydrogenase apoenzyme, as was demonstrated by the detection of dye-linked alcohol dehydrogenase activity after the addition of pqq (pyrroloquinoline quinone). the apoenzyme was purified to homogeneity, and the holoenzyme was characterized. primary alcohols (except methanol), secondary alcohols and aldehydes were substrates, and a broad range of dyes functioned as artificial electron acceptor. optima ... | 1986 | 3521592 |
| high affinity protein-binding and enzyme-inducing activity of methyltrienolone in pseudomonas testosteroni. | the synthetic androgen methyltrienolone (r 1881) was shown to increase steroid delta 1 dehydrogenase activity when added to cultures of pseudomonas testosteroni at concentrations of 10(-10)-10(-8)m. incubation with a soluble extract of p. testosteroni showed that (3h)-r 1881 was bound to a macromolecule with high affinity (kd 0.6 x 10(-9)m) and low capacity (number of binding sites 120 x 10(-15) mol/mg of protein). the (3h)-r 1881-macromolecule complex was partially destroyed following treatment ... | 1986 | 3490081 |
| the laboratory diagnosis of peritonitis during continuous ambulatory peritoneal dialysis. | in 24 episodes of continuous ambulatory peritoneal dialysis associated peritonitis occurring in 21 patients, all dialysis fluids were cloudy and contained at least 100 cells mm-3, mostly polymorphs. gram-positive cocci were seen in centrifuge deposits from only nine of the fluids, but enrichment of 5 ml in liquoid and glucose broth yielded bacteria for all episodes, namely staphylococcus epidermidis (seven episodes), staph. aureus (six), streptococcus mitior (one), moraxella spp (four), acinetob ... | 1986 | 2871078 |
| positioning of a spin-labeled substrate analogue into the structure of delta 5-3-ketosteroid isomerase by combined kinetic, magnetic resonance, and x-ray diffraction methods. | we have shown by kinetic and magnetic resonance measurements that a spin-labeled substrate analogue, spiro[doxyl-2,3'-5' alpha-androstan]-17'beta-ol, binds at the substrate site of crystalline delta 5-3-ketosteroid isomerase (steroid delta-isomerase; ec 5.3.3.1) of pseudomonas testosteroni. the spin-labeled steroid is a linear competitive inhibitor with a ki value (25 +/- 5 microm) that is consistent with dissociation constants obtained by direct binding measurements based on changes in the elec ... | 1987 | 2888482 |
| affinity alkylation of 3-oxo-delta 5-steroid isomerase by steroidal 3 beta-oxiranes: identification of the modified amino acid by reduction with hydroxyborohydride. | the steroidal 3 beta-oxirane (3s)-spiro[5 alpha-androstane-3,2'-oxiran]-17 beta-ol (1 beta) is an active site directed irreversible inhibitor of the 3-oxo-delta 5-steroid isomerase from pseudomonas testosteroni. two steroid-bound peptides (tps1 and tps2) were isolated by high-performance liquid chromatography (hplc) from the trypsin digest of enzyme inactivated with 1 beta. the modified tryptic peptides (residues 14-45 of the enzyme) were further digested with chymotrypsin, each giving rise to a ... | 1987 | 3620446 |
| pseudomonas testosteroni infections: eighteen recent cases and a review of the literature. | pseudomonas testosteroni has been largely overlooked as a potential pathogen in humans. ten cases of infection due to p. testosteroni were identified at a single metropolitan hospital in texas during a three-year period. the organism was most often found in association with anatomic abnormalities of the gastrointestinal tract (six of 10 cases); perforation of the appendix was the commonest abnormality (five cases). the infections were more often polymicrobial (seven cases) than monomicrobial (th ... | 1987 | 3823716 |
| nad(p)+-independent aldehyde dehydrogenase from pseudomonas testosteroni. a novel type of molybdenum-containing hydroxylase. | aldehyde dehydrogenase from pseudomonas testosteroni was purified to homogeneity. the enzyme has a ph optimum of 8.2, uses a wide range of aldehydes as substrates and cationic dyes (wurster's blue, phenazine methosulphate and thionine), but not anionic dyes (ferricyanide and 2.6-dichloroindophenol), nad(p)+ or o2, as electron acceptors. haem c and pyrroloquinoline quinone appeared to be absent but the common cofactors of molybdenum hydroxylases were present. xanthine was not a substrate and allo ... | 1987 | 3609027 |
| inhibition of 3(17)beta-hydroxysteroid dehydrogenase from pseudomonas testosteroni by steroidal a ring fused pyrazoles. | several 2,3- and 3,4-steroidal fused pyrazoles have been investigated as potential inhibitors of nad(p)h-dependent steroid oxidoreductases. these compounds are proven to be potent, specific inhibitors for 3(17) beta-hydroxysteroid dehydrogenase from pseudomonas testosteroni with ki values of 6-100 nm. in contrast, the activities of 3 alpha,20 beta-hydroxysteroid dehydrogenase from streptomyces hydrogenans, steroid 5 alpha-reductase from rat prostate, and 3 alpha-hydroxysteroid dehydrogenase from ... | 1987 | 3040087 |
| cloning of the gene for delta 5-3-ketosteroid isomerase from pseudomonas testosteroni. | we have cloned an approx. 5-kb fragment of pseudomonas testosteroni dna containing the structural gene of delta 5-3-ketosteroid isomerase into the ecori site of the lambda gt11 genome. escherichia coli infected with these recombinant phages produce a polypeptide which is recognized by antiserum raised against the purified isomerase. four of the recombinant lambda gt11 clones contain significant levels of isomerase activity and produce an immunopositive polypeptide of the same apparent mr as the ... | 1987 | 3428616 |
| isolation and sequencing of the gene encoding delta 5-3-ketosteroid isomerase of pseudomonas testosteroni: overexpression of the protein. | we describe the cloning, sequencing, and overexpression of the steroid isomerase (3-oxosteroid delta 5-delta 4-isomerase, ec 5.3.3.1) gene of pseudomonas testosteroni. a genomic library of p. testosteroni total dna constructed from partial ecori digests ligated to a lambda gtwes vector was probed with a 23-base oligonucleotide mixture [atgaac(t)acc(a,t)ccg(c,a)gag(a)cac(t)atgac] corresponding to the nh2-terminal sequence of steroid isomerase. subclones derived from a recombinant phage containing ... | 1987 | 3480517 |
| lyophilized clostridium perfringens 3 alpha- and clostridium bifermentans 7 alpha-hydroxysteroid dehydrogenases: two new stable enzyme preparations for routine bile acid analysis. | preparations of 3 alpha-hydroxysteroid dehydrogenase (ec 1.1.1.50) from clostridium perfringens were successfully lyophilized into a stable powder form. purification of the enzyme was achieved using triazine dye affinity chromatography. c. perfringens 3 alpha-hydroxysteroid dehydrogenase was purified 24-fold using reactive red 120 (procion red) -cross-linked agarose (70% yield). quantitative measurement of bile acids with the purified enzymes, 3 alpha-hydroxysteroid dehydrogenase and 7 alpha-hyd ... | 1988 | 2901274 |
| new naphthalene-degrading marine pseudomonas strains. | over 100 strains that utilized naphthalene as the only carbon and energy source were isolated from samples of marine sediments taken from a heavily polluted area. the isolates were characterized taxonomically and physiologically. most of these strains belonged to the genus pseudomonas, and seven of them did not fit any previous taxonomic description. they differed from type strains in a few biochemical characteristics and in the utilization of aromatic compounds. none had catechol 1,2-dioxygenas ... | 1988 | 3202629 |
| nucleotide sequence of the gene for the delta 5-3-ketosteroid isomerase of pseudomonas testosteroni. | the structural gene for the delta 5-3-ketosteroid isomerase of pseudomonas testosteroni has been sequenced by the dideoxy method. the sequence obtained confirms the amino acid (aa) sequence of benson et al. [j. biol. chem. 246 (1971) 7514-7525] at all but 5 aa residues of the 125-aa polypeptide. amino acid residues 22, 24, 33, and 38, reported to be asparagines by benson et al., are found to be encoded by aspartic acid codons. amino acid residue 77, reported to be a glutamine by benson et al., i ... | 1988 | 3224818 |
| isolation and characterization of a 50 kda testosterone-binding protein from pseudomonas testosteroni. | a testosterone-binding protein (mr = 50,500) has been isolated from the gram-negative bacterium pseudomonas testosteroni. the protein was partially purified by a combination of ion exchange chromatography and chromatofocusing. final purification was achieved by electroelution of the 50 kda protein from sds-polyacrylamide gels. following renaturation from a diluted solution of guanidine-hcl, specific binding of [3h]testosterone to the purified protein was observed. the native protein has a pi of ... | 1989 | 2913397 |
| [a sarcoma-static new species of pseudomonas, pseudomonas jinanensis sp. nov]. | a strain of gram negative bacteria was isolated from the surface soil of wuying hill at jinan, shandong province with gause's medium in 1973. it is a strain of antagonistic bacteria for hysterocervicoma, hepatoma and melanoma of mice screened from 2100 strains of bacteria. it is also antagonistic to staphylococcus aureus, bacillus subtilis and micrococcus. it is a gram negative bacterium with lophotrichous polar flagella. straight rods in shape or with a little slightly curved rods, 0.5-0.6 x 1- ... | 1989 | 2781786 |
| kinetic and ultraviolet spectroscopic studies of active-site mutants of delta 5-3-ketosteroid isomerase. | delta 5-3-ketosteroid isomerase (ec 5.3.3.1) of pseudomonas testosteroni promotes the highly efficient isomerization of delta 5-3-ketosteroids to delta 4-3-ketosteroids by means of a direct and stereospecific transfer of the 4 beta-proton to the 6 beta-position, via an enolic intermediate. an acidic residue responsible for the protonation of the 3-carbonyl function of the steroid and a basic group concerned with the proton transfer have been implicated in the catalytic mechanism. recent nmr stud ... | 1989 | 2706241 |
| degradation of p-toluenesulphonic acid via sidechain oxidation, desulphonation and meta ring cleavage in pseudomonas (comamonas) testosteroni t-2. | pseudomonas (comamonas) testosteroni t-2 completely converted p-toluenesulphonic acid (ts) or p-sulphobenzoic acid (psb) to cell material, co2 and sulphate, with growth yields of about 5 g protein (mol c)-1. psb and sulphite were excreted as transient intermediates during growth in ts-salts medium. all reactions of a catabolic pathway involving sidechain oxidation and cleavage of the sulphonate moiety as sulphite were measurable in the soluble portion of cell extracts. degradation of ts and psb ... | 1989 | 2614395 |
| gentisate 1,2-dioxygenase from pseudomonas. substrate coordination to active site fe2+ and mechanism of turnover. | gentisate 1,2-dioxygenase catalyzes the oxygenolytic ring cleavage of gentisate (2,5-dihydroxybenzoate) between carbons 1 and 2 to form maleylpyruvate. the essential active site fe2+ of the enzyme binds no to yield an epr-active (s = 3/2) complex. hyperfine broadening from 17o (i = 5/2) is observed in the spectrum of the enzyme-nitrosyl complex prepared in 17o-enriched water, demonstrating that water is an iron ligand. association of gentisate with the enzyme-nitrosyl complex causes the broadeni ... | 1990 | 2266121 |
| gentisate 1,2-dioxygenase from pseudomonas. purification, characterization, and comparison of the enzymes from pseudomonas testosteroni and pseudomonas acidovorans. | the 3-hydroxybenzoate inducible gentisate 1,2-dioxygenases have been purified to homogeneity from p. acidovorans and p. testosteroni, the two divergent species of the acidovorans group of pseudomonas. both enzymes exhibit a 40-fold higher specific activity than previous preparations and have an (alpha fe)4 quaternary structure (holoenzyme mr = 164,000 and 158,000, respectively). the enzymes have different amino terminal sequences, amino acid contents, and isoelectric points. each enzyme contains ... | 1990 | 2156846 |
| quinoprotein alcohol dehydrogenase from pseudomonas aeruginosa and quinohemoprotein alcohol dehydrogenase from pseudomonas testosteroni. | 1990 | 2126332 | |
| induction of il-1 during hemodialysis: transmembrane passage of intact endotoxins (lps). | circulating monocytes of patients undergoing chronic hemodialysis are triggered to produce interleukin-1 (il-1) in vivo. intradialytic induction of il-1 is associated with complement activation in patients dialyzed with first-use cellulose membranes. chronic stimulation of il-1 production occurs because of an yet unidentified mechanism in patients dialyzed with high permeability membranes. the present study demonstrates that intact bacterial lipopolysaccharide (lps) molecules may cross cuprophan ... | 1990 | 2127434 |
| protocatechuate 4,5-dioxygenase from pseudomonas testosteroni. | 1990 | 2280721 | |
| inhibition of microbial cholesterol oxidases by dimethylmorpholines. | cholesterol oxidase is a potentially important enzyme in steroid transformations, catalysing the conversion of 3-hydroxy-5-ene steroids to 3-keto-4-ene derivatives via a 3-keto-5-ene intermediate. morpholine derivatives, especially fenpropimorph and tridemorph, were found to block selectively the isomerisation activity of cholesterol oxidases isolated from nocardia erythropolis, streptomyces sp., pseudomonas testosteroni and schizophyllum commune. these enzymes differ strongly in physical charac ... | 1990 | 2308321 |
| cloning and expression of genes involved in 4-chlorobiphenyl transformation by pseudomonas testosteroni: homology to polychlorobiphenyl-degrading genes in other bacteria. | the genes of pseudomonas testosteroni strain b-356, specifying the transformation of 4-chlorobiphenyl (4-cb) into 4-chlorobenzoic acid (4-cba) were cloned into pseudomonas putida kt2440 using a broad-host-range cosmid, ppsa842. of 10,000 clones tested, four were able to transform 4-cb. gas chromatographic and mass spectrometric analysis of the catabolic products from two of the 4-cb-transforming clones carrying the hybrid plasmids, pda1 and pda2, demonstrated that pda1 carried a complete set of ... | 1990 | 2311936 |
| beta-hydroxysteroid dehydrogenase: activity in microemulsion and extraction from pseudomonas testosteroni cells with microemulsion. | the stability of purified beta-hydroxysteroid dehydrogenase activity measured as a function of time was good in buffered cationic and non-ionic microemulsions. the use of 1-pentanol and 1-hexanol in place of 1-butanol as cosurfactant gave increased activity and stability. the nad+ michaelis constant was 0.22 mm in buffer and 3.5 mm in waterpool concentration in microemulsion. proteins, among them beta-hydroxysteroid dehydrogenase, were extracted from pseudomonas testosteroni with cationic microe ... | 1990 | 2116917 |
| chlorinated biphenyl mineralization by individual populations and consortia of freshwater bacteria. | comparative studies were performed to investigate the contribution of microbial consortia, individual microbial populations, and specific plasmids to chlorinated biphenyl biodegradation among microbial communities from a polychlorinated biphenyl-contaminated freshwater environment. a bacterial consortium, designated lps10, was shown to mineralize 4-chlorobiphenyl (4cb) and dehalogenate 4,4'-dichlorobiphenyl. the lps10 consortium involved three isolates: pseudomonas testosteroni (lps10a), which m ... | 1990 | 2117875 |
| microbial degradation of the morphine alkaloids: identification of morphine as an intermediate in the metabolism of morphine by pseudomonas putida m10. | a strain of pseudomonas putida was isolated by selective enrichment with morphine that was capable of utilising morphine as a primary source of carbon and energy for growth. experiments with whole cells showed that both morphine and codeine, but not thebaine, could be utilised. a novel nadp-dependent dehydrogenase, morphine dehydrogenase, was purified from crude cell extracts and was shown to be capable of oxidising morphine and codeine to morphinone and codeinone, respectively. this nadp-depend ... | 1990 | 1701625 |
| carbonyl reduction of metyrapone in human liver. | carbonyl reduction was investigated in cytosolic and microsomal fractions of human liver using the ketone metyrapone as a substrate. the cytosolic enzyme has a stronger preference for nadph over nadh than the microsomal enzyme: the former shows only 14% of the nadph-supported activity while the latter exhibits 36% activity with nadh. barbitone and quercitrin, the classic inhibitors of carbonyl reductases, do not affect metyrapone reduction in either fraction. dicumarol and indomethacin, the spec ... | 1991 | 1722672 |
| subcloning of bph genes from pseudomonas testosteroni b-356 in pseudomonas putida and escherichia coli: evidence for dehalogenation during initial attack on chlorobiphenyls. | the bpha, -b, -c, and -d genes from pseudomonas testosteroni b-356 were mapped to a 5.5-kb dna fragment of cloned plasmids pda1 and pda2 by use of deletion and insertion mutants of these plasmids. the expression of each of these genes was evaluated in escherichia coli and in pseudomonas putida, and it was found that the bphc and bphd genes are well expressed in both e. coli and p. putida cells while the bpha and bphb genes are very poorly expressed in e. coli, even when placed downstream of a ta ... | 1991 | 1746948 |
| bioconversion of 2-hydroxy-6-oxo-6-(4'-chlorophenyl)hexa-2,4-dienoic acid, the meta-cleavage product of 4-chlorobiphenyl. | bacterial conversion of 4-chlorobiphenyl (4-cb) usually proceeds through a pathway involving an initial oxidation of the unsubstituted ring in the 2,3 position followed by a 1,2 meta-cleavage. the meta-cleavage product (mcp) is converted through a single hydrolysis step into chlorobenzoic acid. however, several other acidic metabolites that were not expected as part of this pathway have already been described. in this paper, we used strains of pseudomonas putida carrying cloned genes from pseudo ... | 1991 | 1919512 |
| broad host-range vector for efficient expression of foreign genes in gram-negative bacteria. | a broad host-range expression plasmid was constructed comprising the incq replicon, the reca promoter from escherichia coli and the g10-l ribosome binding site (rbs) derived from bacteriophage t7. the structural genes for porcine somatotropin (pst) and e. coli beta-galactosidase (lacz) were used to monitor gene expression in a diverse collection of gram-negative bacterial hosts: escherichia coli, pseudomonas aeruginosa, pseudomonas syringae, pseudomonas putida, pseudomonas fluorescens, pseudomon ... | 1991 | 1367537 |
| 4-sulphobenzoate 3,4-dioxygenase. purification and properties of a desulphonative two-component enzyme system from comamonas testosteroni t-2. | cell-free extracts of comamonas testosteroni t-2 grown in toluene-p-sulphonate/salts medium catalyse the conversion of p-sulphobenzoate (psb) into protocatechuate and sulphite by an nadh-requiring and fe2(+)-activated dioxygenase. anion-exchange chromatography of extracts yielded red (a) and yellow (b) protein fractions, both of which were necessary for dioxygenative activity. further purification of each fraction by hydrophobic interaction chromatography and gel filtration led to two homogeneou ... | 1991 | 2012609 |
| pseudomonas 3 beta-hydroxysteroid dehydrogenase. primary structure and relationships to other steroid dehydrogenases. | the 3 beta-hydroxysteroid dehydrogenase of pseudomonas testosteroni commercially available was purified by an fplc step and submitted to sequence determination by peptide analysis. the structure obtained reveals a 253-residue polypeptide chain, with an n-terminal, free alpha-amino group, and a low cysteine content. comparisons with other hydroxysteroid dehydrogenases recently characterized reveal distant similarities with prokaryotic and, to some extent, also eukaryotic forms of separate specifi ... | 1991 | 2026158 |
| catalytic mechanism of an active-site mutant (d38n) of delta 5-3-ketosteroid isomerase. direct spectroscopic evidence for dienol intermediates. | the delta 5-3-ketosteroid isomerase (ec 5.3.3.1) of pseudomonas testosteroni catalyzes the conversion of androst-5-ene-3,17-dione to androst-4-ene-3,17-dione by a stereospecific transfer of the 4 beta-proton to the 6 beta-position. the reaction involves two steps: (a) a rate-limiting concerted enolization, comprising protonation of the 3-carbonyl oxygen by the phenolic hydroxyl group of tyr-14 and abstraction of the 4 beta-proton by the carboxylate group of asp-38, and (b) rapid reketonization o ... | 1991 | 2036366 |
| 4-toluene sulfonate methyl-monooxygenase from comamonas testosteroni t-2: purification and some properties of the oxygenase component. | comamonas testosteroni t-2 synthesizes an inducible enzyme system that oxygenates 4-toluene sulfontate (ts) to 4-sulfobenzyl alcohol when grown in ts-salts medium. we purified this ts methyl-monooxygenase system (tsmos) and found it to consist of two components. a monomeric, iron-sulfur flavoprotein (component b), which has been shown to act as a reductase in the 4-sulfobenzoate dioxygenase system of this organism (h. h. locher, t. leisinger, and a. m. cook, biochem. j. 274:833-842, 1991), carri ... | 1991 | 2050632 |
| cloning, sequencing, and expression of the pseudomonas testosteroni gene encoding 3-oxosteroid delta 1-dehydrogenase. | pseudomonas testosteroni atcc 17410 is able to grow on testosterone. this strain was mutagenized by tn5, and 41 mutants defective in the utilization of testosterone were isolated. one of them, called mutant 06, expressed 3-oxosteroid delta 1- and 3-oxosteroid delta 4-5 alpha-dehydrogenases only at low levels. the dna region around the tn5 insertion in mutant 06 was cloned into puc19, and the 1-kbp ecori-bamhi segment neighbor to the tn5 insertion was used to probe dna from the wild-type strain. ... | 1991 | 1657885 |
| molecular cloning and expression of the beta-hydroxysteroid dehydrogenase gene from pseudomonas testosteroni. | the structural gene (hsd) of the pseudomonas testosteroni encoding the 17 beta-hydroxysteroid dehydrogenase has been cloned using the cosmid vector pvk102. escherichia coli carrying recombinant clones of hsd, isolated by immunological screening, were able to express the biologically active enzyme, as measured by the conversion of testosterone into androstenedione. subcloning experiments, restriction and deletion analysis, and site-directed insertion mutagenesis showed that the hsd gene is locate ... | 1991 | 1657714 |
| studies of the catalytic mechanism of an active-site mutant (y14f) of delta 5-3-ketosteroid isomerase by kinetic deuterium isotope effects. | delta 5-3-ketosteroid isomerase (ec 5.3.3.1) from pseudomonas testosteroni catalyzes the conversion of androst-5-ene-3,17-dione to androst-4-ene-3,17-dione by a stereoselective transfer of the 4 beta-proton to the 6 beta-position. the rate-limiting step has been shown to be the concerted enolization of the enzyme-bound substrate comprising protonation of the 3-carbonyl oxygen by tyr-14 and abstraction of the 4 beta-proton by asp-38 [xue, l., talalay, p., & mildvan, a. s. (1990) biochemistry 29, ... | 1991 | 1932008 |
| transfer and expression of the catabolic plasmid pbrc60 in wild bacterial recipients in a freshwater ecosystem. | 3-chlorobenzoate (3cba)-degrading bacteria were isolated from the waters and sediments of flowthrough mesocosms dosed with various concentrations of 3cba and inoculated with a 3cba-degrading alcaligenes sp., strain br60. bacteria capable of 3cba degradation which were distinct from br60 were isolated. they carried pbrc60, a plasmid introduced with alcaligenes sp. strain br60 that carries a transposable element (tn5271) encoding 3cba degradation. the isolates expressed these genes in different wa ... | 1991 | 16348493 |
| dye-linked dehydrogenase activities for formate and formate esters in amycolatopsis methanolica. characterization of a molybdoprotein enzyme active with formate esters and aldehydes. | cell-free extracts of methanol-grown amycolatopsis methanolica contain dye-linked dehydrogenase activities for formate and methyl formate. fractionation of the extracts revealed that the (unstable) activity for formate resides in membrane particles, while that for methyl formate belongs to a soluble enzyme that was purified and characterized. the enzyme, indicated as formate-ester dehydrogenase, appeared to be a molybdoprotein (4 fe, 3 or 4 s, 1 mo and 1 fad were found for each enzyme molecule), ... | 1992 | 1597191 |
| modelling of mixed chemostat cultures of an aerobic bacterium, comamonas testosteroni, and an anaerobic bacterium, veillonella alcalescens: comparison with experimental data. | a mathematical model of mixed chemostat cultures of the obligately aerobic bacterium comamonas testosteroni and the anaerobic bacterium veillonella alcalescens grown under dual limitation of l-lactate and oxygen was constructed. the model was based on michaelis-menten-type kinetics for the consumption of substrates, with noncompetitive inhibition of v. alcalescens by o2. the growth characteristics of the aerobic and anaerobic organisms were determined experimentally with pure cultures of the ind ... | 1992 | 1622213 |
| identification of xenobiotic-degrading isolates from the beta subclass of the proteobacteria by a polyphasic approach including 16s rrna partial sequencing. | nineteen gram-negative, aerobic, biodegradative isolates were identified by using a polyphasic taxonomic approach. the presence of the specific polyamine 2-hydroxyputrescine and the presence of a ubiquinone with eight isoprenoid units in the side chain (ubiquinone q-8) allowed allocation of these organisms to the beta subclass of the proteobacteria. on the basis of the results of additional characterization experiments (i.e., api 20ne tests, determinations of soluble protein patterns, and dna-dn ... | 1992 | 1371062 |
| a novel sulfatase from pseudomonas testosteroni hydrolyzing lithocholic acid sulfate. | pseudomonas testosteroni atcc 11996 was found to produce a novel bile acid sulfate sulfatase that hydrolyzes the sulfate ester bond in lithocholic acid sulfate (lca-s). the enzyme synthesis was induced by several kinds of bile acids including lca-s. mn2+ functioned as an essential component for the enzyme synthesis and so4(2-) suppressed it. this sulfatase hydrolyzes lca-s to isolithocholic acid and sulfuric acid with inversion of alpha- to beta-configuration of the hydroxyl group at the third p ... | 1992 | 1369058 |
| substrate polarization by residues in delta 5-3-ketosteroid isomerase probed by site-directed mutagenesis and uv resonance raman spectroscopy. | delta 5-3-ketosteroid isomerase (ksi: ec 5.3.3.1) of pseudomonas testosteroni catalyzes the isomerization of delta 5-3-ketosteroids to delta 4-3-ketosteroids by the stereospecific transfer of the steroid 4 beta-proton to the 6 beta-position, using tyr-14 as a general acid and asp-38 as a base. ultraviolet resonance raman (uvrr) spectra have been obtained for the catalytically active double mutant y55f + y88f, which retains tyr-14 as the only tyrosine residue (referred to as the y14(0) mutant), a ... | 1992 | 1339027 |
| functional and immunological relationships between metyrapone reductase from mouse liver microsomes and 3 alpha-hydroxysteroid dehydrogenase from pseudomonas testosteroni. | 3 alpha-hydroxysteroid dehydrogenase (3 alpha-hsd) from pseudomonas testosteroni was shown to reduce the xenobiotic carbonyl compound metyrapone (mpon). reversely, mpon reductase purified from mouse liver microsomes and previously characterized as aldehyde reductase, was competitively inhibited by 3 alpha-hsd steroid substrates. for mpon reduction both enzymes can use either nadh or nadph as co-substrate. immunoblot analysis after native and sds gel electrophoresis of 3 alpha-hsd gave a specific ... | 1992 | 1551429 |
| homologies between enzymes involved in steroid and xenobiotic carbonyl reduction in vertebrates, invertebrates and procaryonts. | evidence is reported for the existence of a structurally and functionally related and probably evolutionarily conserved class of membrane-bound liver carbonyl reductases/hydroxysteroid dehydrogenases involved in steroid and xenobiotic carbonyl metabolism. carbonyl reduction was investigated in liver microsomes of 8 vertebrate species, as well as in insect larvae total homogenate and in purified 3 alpha-hydroxysteroid dehydrogenase preparations of the procaryont pseudomonas testosteroni, using th ... | 1992 | 1472459 |
| outer membranes of gram-negative bacteria are permeable to steroid probes. | the permeability of bacterial outer membranes was assayed by coupling the influx of highly hydrophobic probes, 3-oxosteroids, with their subsequent oxidation catalysed by 3-oxosteroid delta 1-dehydrogenase, expressed from a gene cloned from pseudomonas testosteroni. in salmonella typhimurium producing wild-type lipopolysaccharide, the permeability coefficients for uncharged steroids were 0.45 to 1 x 10(-5) cm s-1, and the diffusion appeared to occur mainly through the lipid bilayer domains of th ... | 1992 | 1640833 |
| change of speciation of cu(ii) in growth medium due to uptake of ammonia by pseudomonas testosteroni during growth. | growth of pseudomonas testosteroni in a medium containing 1 mm cu(ii) causes a color change from blue to green. the spectrum of the supernatant solution from the blue culture shows an absorption at 660 nm, identical to that of 1 mm [cu(ii)] in the medium. the green supernatant solution shows a uv absorption, which tails into the visible and so is responsible for the green color, and a d-d absorption at 720 nm. the absorption at 660 nm for the blue supernatant solution is probably due to [cu(nh3) ... | 1992 | 1369191 |
| specific activation of a tyrosine----glycine mutant of delta 5-3-ketosteroid isomerase by phenols. | a key unknown still to be explored concerning the mechanism of delta 5-3-ketosteroid isomerase from pseudomonas testosteroni is the extent of the proton transfer between tyrosine-14 of the enzyme and the c-3 carbonyl oxygen of the steroid substrate. this report is a preliminary study of a system we are developing to allow us eventually to use a brønsted analysis to measure this transfer. we describe the construction of an expression vector and tyrosine-14----glycine-14 mutant of the enzyme and i ... | 1992 | 1590799 |
| kinetics and stability of delta 5-3-ketosteroid isomerase from pseudomonas testosteroni in the system of reverse micelles of aerosol ot in isooctane. | partially purified delta 5-3-ketosteroid isomerase (ksi) from pseudomonas testosteroni was studied kinetically after solubilization in reverse micelles of aerosol ot (aot) in isooctane and water, as regards its application to biotechnology. with delta 5,10-estren-17 beta-ol-3-one as a substrate, ksi displays an enzyme activity in the micellar system but a low stability. in the presence of urea, the enzyme is, however, stable. kinetic parameters of the stabilized enzyme are highly sensitive to bo ... | 1992 | 1381618 |
| effects of chlorobenzoate transformation on the pseudomonas testosteroni biphenyl and chlorobiphenyl degradation pathway. | bacterial conversion of biphenyl (bp) and chlorobiphenyls (cbps) to benzoates and chlorobenzoates (cbas) proceeds by introduction of molecular oxygen at the 2,3 position, followed by a 1,2-meta cleavage of the molecule. complete mineralization of cbps requires the presence of two sets of genes, one for the transformation fo cbps into cbas and a second for the degradation of cbas. it has been shown previously that removal of the cbas produced from the degradation of cbps is essential for efficien ... | 1992 | 1610172 |
| use of a simplified cell blot technique and 16s rrna-directed probes for identification of common environmental isolates. | a simple technique in which rrna-targeted oligodeoxynucleotide probes are used to identify bacteria immobilized on membranes is described. by using colony lifts, bacteria are directly transferred from plates to untreated nitrocellulose membranes. alternatively, cells resuspended from colonies can be applied to membranes by using a vacuum manifold under high-salt conditions. blotted cells are baked and hybridized under stringent conditions by using standard protocols. treatment of blotted cells w ... | 1993 | 7504429 |
| kinetics and mechanism of heterogeneous hydrolysis of poly[(r)-3-hydroxybutyrate] film by pha depolymerases. | the kinetics and mechanism of enzymatic degradation on the surface of poly[(r)-3-hydroxybutyrate] (p[(r)-3hb]) film have been studied using three types of extracellular poly(hydroxyalkanoate) (pha) depolymerases from alcaligenes faecalis, pseudomonas picketti and comamonas testosteroni. the monomer and dimer of 3-hydroxybutyric acid were produced during the course of the enzymatic degradation of p[(r)-3hb] film, and the rate of production was determined by monitoring the increase in absorbance a ... | 1993 | 8110658 |
| cloning, dna sequencing and expression of (3-17)beta hydroxysteroid dehydrogenase from pseudomonas testosteroni. | we describe the cloning, sequencing and overexpression of the (3-17)beta hydroxysteroid dehydrogenase gene of pseudomonas testosteroni. a genomic library of ps. testosteroni total dna constructed from sauiiia digests ligated to a lambda gt11 vector was probed with polyclonal antibody raised against purified enzyme. subclones derived from a recombinant phage containing a 2661-base-pair insert were sequenced and found to contain an open reading frame of 765 base pairs that corresponds to a protein ... | 1993 | 8382516 |
| telluria mixta (pseudomonas mixta bowman, sly, and hayward 1988) gen. nov., comb. nov., and telluria chitinolytica sp. nov., soil-dwelling organisms which actively degrade polysaccharides. | pseudomonas mixta (type strain, acm 1762 [= atcc 49108], an actively dextranolytic species that possesses both lateral and polar flagella, was compared with the strictly aerobic, rod-shaped, chitinolytic bacterium "pseudomonas chitinolytica" acm 3522t (= cncm i-804) (t = type strain), which has a similar flagellation pattern, by performing phenotypic characterization and dna-dna hybridization studies and by analyzing dna base compositions and 16s rrna sequences. our results indicated that "p. ch ... | 1993 | 8427803 |
| uptake of 4-toluene sulfonate by comamonas testosteroni t-2. | the mechanism of transport of the xenobiotic 4-toluene sulfonate (ts) in comamonas testosteroni t-2 was investigated. rapid uptake of ts was observed only in cells grown with ts or 4-methylbenzoate as a carbon and energy source. initial uptake rates under aerobic conditions showed substrate saturation kinetics, with an apparent affinity constant (kt) of 88 microm and a maximal velocity (vmax) of 26.5 nmol/min/mg of protein. uptake of ts was inhibited completely by uncouplers and only marginally ... | 1993 | 8432701 |
| improved purification of steroid 1:2-dehydrogenase from nocardia opaca and partial characterization of its cloned gene sequence. | we have purified a steroid-inducible 1:2-dehydrogenase from nocardia opaca. the final enzyme preparation was purified 120-fold with a recovery of 38%. the n-terminal amino acid sequence was determined to be: met-gln-asp-trp-thr-ser-glu-(cys)-asp-val-leu-val-val-gly-. from the genomic library of nocardia opaca in the plasmid puc19, a clone designated as pstd23 containing a 0.9 kb kpni-psti fragment was found to hybridize with an oligonucleotide probe corresponding to the first six amino acids fro ... | 1993 | 7916596 |
| microbial metabolism of quinoline and related compounds. xvii. degradation of 3-methylquinoline by comamonas testosteroni 63. | a bacterial strain which utilizes 3-methylquinoline as sole source of carbon, nitrogen and energy was isolated from activated sludge. on the basis of its morphological and physiological characteristics, this isolate was classified as comamonas testosteroni. four metabolites of 3-methylquinoline degradation were isolated from the culture supernatant and identified as 3-methyl-2-oxo-1,2-dihydroquinoline, 6-hydroxy-3-methyl-2-oxo-1,2-dihydroquinoline, 5,6-dihydroxy-3-methyl-2-oxo-1,2-dihydroquinoli ... | 1993 | 8489738 |
| factors affecting pcb degradation by an implanted bacterial strain in soil microcosms. | pseudomonas testosteroni b-356 was able to degrade approximately 50% of the aroclor 1242 mixture in shaken culture. the aims of the present study were to evaluate the capabilities of this bacterial strain to degrade pcbs in soil microcosms and to identify some of the factors likely to favor the degradative performance of the implanted bacteria. the presence of biphenyl as cosubstrate was the most important factor affecting pcb degradation in soil. however, because biphenyl was rapidly depleted i ... | 1993 | 8358671 |
| environments and mechanistic roles of the tyrosine residues of delta 5-3-ketosteroid isomerase. | delta 5-3-ketosteroid isomerase (ec 5.3.3.1) from pseudomonas testosteroni converts delta 5-3-ketosteroids to delta 4-3-ketosteroids by a stereoselective and conservative transfer of the 4 beta-proton to the 6 beta-position. the 10(9.5)-fold enzymatic rate acceleration can be attributed to a concerted rate-limiting enolization in which tyr-14 and asp-38, positioned orthogonally, act as general acid and base, respectively. the pka value of the phenolic hydroxyl group of tyr-14 of the y55f/y88f do ... | 1993 | 8439542 |
| carbonyl reduction by 3 alpha-hsd from comamonas testosteroni--new properties and its relationship to the scad family. | 1993 | 8493916 | |
| degradation of delor 103, a technical mixture of polychlorinated biphenyls, by selected bacteria. | ability to utilize a technical mixture of polychlorinated biphenyls (pcb), delor 103, as the sole carbon source, has been tested in 14 bacterial strains. for the five best growing strains (alcaligenes latus, alcalgenes eutrophus, comamonas testosteroni, micrococcus varians and pseudomonas putida), the dependence of the degradation of individual pcb congeners on the number of chlorine substituents is discussed. | 1993 | 24420291 |
| purification and properties of bile acid sulfate sulfatase from pseudomonas testosteroni. | the bile acid sulfate sulfatase (bss) produced by pseudomonas testosteroni was purified and characterized. chromatofocusing behavior and amino acid sequence over twelve amino acid residues from n-terminus of the enzyme indicated that bss was composed of two isoforms of which molecular weights were 125,000 and 103,000. each isoform was a homodimer of a subunit of which molecular weight was 53,000 or 51,000, respectively. the optimum ph was 8.5 and bss was stable at ph 5.8-8.0. the thermostability ... | 1994 | 7764976 |
| mechanism of the reaction catalyzed by delta 5-3-ketosteroid isomerase of comamonas (pseudomonas) testosteroni: kinetic properties of a modified enzyme in which tyrosine 14 is replaced by 3-fluorotyrosine. | tyrosine 14 of delta 5-3-ketosteroid isomerase plays an important role in the function of the enzyme, since its replacement by phenylalanine results in a decrease in kcat by a factor of 10(-4.7). this result and the fact that this residue resides in the enzyme's substrate binding site and is in close proximity to c-2 of the bound steroid suggests that it functions as an electrophile in the catalytic mechanism by protonation of or hydrogen bonding to the c-3 carbonyl oxygen of the substrate. in o ... | 1994 | 8117732 |
| regulation of chloro- and methylphenol degradation in comamonas testosteroni jh5. | comamonas testosteroni jh5 was isolated from a mixed bacterial culture enriched on different chloro- and methylphenols. the strain completely mineralized a mixture consisting of 4-chlorophenol (4-cp) and 4-methylphenol (4-mp). during degradation of the mixture, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, 4-hydroxybenzoic acid, and 4-chlorocatechol were detected as short-lived intermediates. mineralization of 4-cp and that of 4-mp occurred successively and were accompanied by diauxic growth, ... | 1994 | 8074514 |
| identification and mapping of the gene translation products involved in the first steps of the comamonas testosteroni b-356 biphenyl/chlorobiphenyl biodegradation pathway. | in this study, we have mapped comamonas testosteroni b-356 genes encoding enzymes for the conversion of biphenyl and 4-chlorobiphenyl into the corresponding meta-cleavage compounds onto a 6.3-kb dna fragment, and we have determined the subunit composition of the enzymes involved in this pathway. the various proteins encoded by this 6.3-kb dna fragment and by subclones derived from it were overexpressed and selectively labelled using the t7 polymerase promoter system in escherichia coli. they wer ... | 1994 | 7954108 |
| terephthalate 1,2-dioxygenase system from comamonas testosteroni t-2: purification and some properties of the oxygenase component. | comamonas testosteroni t-2, grown in terephthalate (ter)-salts medium, synthesizes inducible enzymes that convert ter to (1r,2s)-dihydroxy-3,5-cyclohexadiene-1,4-dicarboxylic acid (dcd) and protocatechuate (pc). anion-exchange chromatography of cell extracts yielded two sets of fractions, r and z, that were necessary for oxygenation of ter to dcd; we termed this activity the ter dioxygenase system (terdos). an nad(+)-dependent dcd dehydrogenase, which converted dcd to pc, overlapped all fraction ... | 1994 | 7961417 |
| description of the kinetic mechanism and the enantioselectivity of quinohaemoprotein ethanol dehydrogenase from comamonas testosteroni in the oxidation of alcohols and aldehydes. | initial rate studies were performed on the oxidation of (racemic) alcohols as well as aldehydes by quinohaemoprotein ethanol dehydrogenase, type 1, from comamonas testosteroni with potassium ferricyanide as electron acceptor. the data could be fitted with an equation derived for a mechanism (hexa-uni ping-pong) in which alcohols are oxidized to the corresponding carboxylic acids and the intermediate aldehyde becomes released from the enzyme. however, for some substrates it was necessary to assum ... | 1994 | 8001568 |
| characterization of the three tyrosine residues of delta 5-3-ketosteroid isomerase by time-resolved fluorescence and circular dichroism. | delta 5-3-ketosteroid isomerase (ec 5.3.3.1) of pseudomonas testosteroni converts delta 5-3-ketosteroids to delta 4-3-ketosteroids via an enolic intermediate. site-specific mutagenesis has identified tyr-14 and asp-38 as the catalytically essential general acid and base, respectively. three tyrosine residues (tyr-14, tyr-55, and tyr-88) are the only significant fluorophores in the wild-type isomerase. recent studies of the steady-state fluorescence of the wild-type enzyme and all six mutant enzy ... | 1994 | 8003507 |
| extent of proton transfer in the transition states of the reaction catalyzed by the delta 5-3-ketosteroid isomerase of comamonas (pseudomonas) testosteroni: site-specific replacement of the active site base, aspartate 38, by the weaker base alanine-3-sulfinate. | previous studies of the mechanism of the steroid isomerase of comamonas (pseudomonas) testosteroni have identified aspartate 38 as the proton porter which transfers the substrate's 4 beta proton to the 6 beta position of the product. consequently, aspartate 38 functions as a base in the deprotonation of the substrate to form a dienol or dienolate intermediate, which then undergoes reprotonation from protonated aspartate 38 at c-6 beta to give the product. we have tried to characterize the transi ... | 1994 | 8117731 |
| enzymatic and nonenzymatic polarizations of alpha,beta-unsaturated ketosteroids and phenolic steroids. implications for the roles of hydrogen bonding in the catalytic mechanism of delta 5-3-ketosteroid isomerase. | ketosteroids (e.g., 19-nortestosterone) and phenolic steroids (e.g., 17 beta-estradiol and 17 beta-dihydroequilenin), which are potent competitive inhibitors of delta 5-3-ketosteroid isomerase (isomerase, ec 5.3.3.1) of pseudomonas testosteroni, undergo significant polarization upon binding to the active site of the enzyme. the 10 nm red shift of the uv absorption maximum of the enone chromophore of 19-nortestosterone, which occurs in the enzyme-steroid complex, resembles that observed when this ... | 1995 | 7819234 |
| regulation of the hema gene during 5-aminolevulinic acid formation in pseudomonas aeruginosa. | the general tetrapyrrole precursor 5-aminolevulinic acid is formed in bacteria via two different biosynthetic pathways. members of the alpha group of the proteobacteria use 5-aminolevulinic acid synthase for the condensation of succinyl-coenzyme a and glycine, while other bacteria utilize a two-step pathway from aminoacylated trna(glu). the trna-dependent pathway, involving the enzymes glutamyl-trna reductase (encoded by hema) and glutamate-1-semialdehyde-2,1-aminomutase (encoded by heml), was d ... | 1995 | 7883699 |