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At least 649 records · Page 36Linked to original sources
PubMed1989

Poly-beta-hydroxybutyrate biosynthesis in Alcaligenes eutrophus H16. Characterization of the genes encoding beta-ketothiolase and acetoacetyl-CoA reductase.

The poly-beta-hydroxybutyrate biosynthetic thiolase gene from Zoogloea ramigera was used as a hybridization probe to screen restriction digests of Alcaligenes eutrophus H16 DNA. Specific hybridization signals were obtained and two fragments (a 2.3-kilobase PstI fragment and a 15-kilobase EcoRI fragment) cloned in the Escherichia coli vector pUC8 (plasmids pAeT3/pAeT10 and pAeT29, respectively). Biochemical analysis of lysates of E. coli cells containing each plasmid identified significant levels of beta-ketothiolase and acetoacetyl-CoA reductase enzyme activities in lysates of E. coli cells containing plasmids pAeT10 or pAeT29. Nucleotide sequence analysis of the pAeT10 insert identified two open reading frames which encode polypeptides of Mr = 40,500 and Mr = 26,300 corresponding to the structural genes for beta-ketothiolase (phbA) and acetoacetyl-CoA reductase (phbB), respectively. Amino acid sequence homologies between the two bacterial and two mammalian thiolases are discussed with respect to the chain length specificity exhibited by the different thiolase enzymes.

Acetyl-CoA C-Acyltransferase↗
PubMed1989

Poly-beta-hydroxybutyrate (PHB) biosynthesis in Alcaligenes eutrophus H16. Identification and characterization of the PHB polymerase gene (phbC).

The phbC gene encoding the third enzyme of the poly-beta-hydroxybutyrate biosynthetic pathway, poly-beta-hydroxybutyrate polymerase, in Alcaligenes eutrophus H16 has been identified by the complementation of poly-beta-hydroxybutyrate negative mutants of A. eutrophus H16. These results demonstrate that the three enzymes of the poly-beta-hydroxybutyrate biosynthetic pathway are organized phbC-phbA-phbB. Expression of all three genes in Escherichia coli results in a significant level (50% dry cell weight) of poly-beta-hydroxybutyrate production. phbC encodes a polypeptide of Mr = 63,900 which has a hydropathy profile distinct from typical membrane proteins indicating that poly-beta-hydroxybutyrate biosynthesis probably does not involve a membrane complex.

Alcaligenes↗
PubMed1989

A 2.0-A structure of the blue copper protein (cupredoxin) from Alcaligenes faecalis S-6.

The structure of a blue copper protein, cupredoxin, from the potent denitrifying bacterium Alcaligenes faecalis S-6, has been determined and refined against 2 A x-ray diffraction data. The agreement between observed and calculated structure factors is 0.159, and estimated errors in coordinates are 0.09-0.15 A. The protein folds in a beta sandwich similar to plastocyanin and azurin and includes features such as a "kink" and a "tyrosine loop" which have been noted previously for these proteins as well as immunoglobulins. The copper is bound by four ligands, in a distorted tetrahedral arrangement, with Cu-S gamma = 2.07 A (Cys-78), Cu-N delta 1 = 2.10 and 2.21 for His-40 and His-81, and Cu-S delta = 2.69 A (Met-86). Two of the ligands are further oriented by hydrogen bonds either to other side chains (Asn-9 to His-40), backbone atoms (NH...S) or a water molecule (to His-40). The methionine ligand has no extra constraints. The C-terminal loop containing three of the ligands is hydrogen-bonded to the strand containing His-40 by hydrogen bonds between the conserved residues Thr-79 and Asn-41. The pronounced dichroism of the crystal is a result of the orientation of the normal to the C beta-S gamma-Cu plane parallel to the crystallographic 6-fold axis.

Alcaligenes↗
PubMed1986

Plasmid DNA of virulent Alcaligenes faecalis.

Alcaligenes faecalis strains originating from chickens and from epizootics of coryza in turkeys were screened for antibiotic susceptibility and for the presence of plasmid DNA. Seven of 35 strains contained plasmid DNA ranging in size from 10.5 to approximately 32 megadaltons. All of the strains isolated from turkeys were virulent in turkey poults, but only the plasmid-containing strains were resistant to sulfonamides and streptomycin. Four of the plasmid-containing strains were also resistant to tetracycline. Five different plasmids representing at least 2 different incompatibility groups were identified in the 7 plasmid-bearing A faecalis strain.

Alcaligenes↗
PubMed1988

Refinement of the structure of pseudoazurin from Alcaligenes faecalis S-6 at 1.55 A resolution.

The crystal structure of the redox protein pseudoazurin (123 amino acid residues; molecular weight 13,000 daltons) from Alcaligenes faecalis has been refined by fast Fourier restrained least-squares minimization. Cycles of rebuilding were carried out to escape from local minima. Individual isotropic temperature factor values were refined separately for all atoms. The R factor was reduced from 0.400 (for 2647 reflections in the 6.0-2.8 A resolution range) to 0.180 (for all 19,770 reflections in the 9.0-1.55 A resolution range) with a final estimated accuracy in atomic positions of 0.15 A. The final model comprises 917 protein atoms and 93 solvent molecules. The root-mean-square shift of the main-chain atoms between the final and the initial model is 0.94 A (maximum shift 1.8 A). Most of the larger shifts were the result of rebuilding on the graphics system. The average atomic temperature factor, B, is 23.0 A2 for all atoms. Side-chain atoms with high B values were omitted, and their positions checked from difference maps. The three carboxy-terminal residues were omitted from the final model as no single conformation could be assigned from the observed electron density. All other protein atoms were included.

Alcaligenes↗
PubMed1988

Identification of Pseudomonas, Flavobacterium, and Alcaligenes with the API 20 NE system.

The API 20 NE system is designed for the rapid identification of Gram negative rods other than Enterobacteriaceae. It has been compared to conventional methods in the characterization of 404 strains representative of 23 species belonging to 3 genera: Pseudomonas (236 strains), Flavobacterium (133 strains) and Alcaligenes (35 strains). The system consists of 9 enzymatic tests and 12 carbohydrate assimilation tests. A 7-digit numerical profile is obtained by an octal coding of the test results. The strains are identified by comparing the numerical profile to those of species listed in a profile index. The API 20 NE tests showed 99-100% correlation with corresponding conventional methods except in indole production (96.5%) for which we observed 14 false negative reactions in testing F. meningosepticum and CDC groups IIb and IIf. The API 20 NE system presented 94.1% identification to the species level and 96.3% to the genus level in agreement with conventional biochemical methods. Among the correctly identified strains, 7.4% (30) gave an erroneous or unlisted numerical profile in the first version of the Profile Index and required the use of the computer. Among the misidentified strains 2%, (8) were assigned to the right genus but wrong species, 0.7% (3) were placed in the wrong genus and 1% (4) gave a doubtful profile with one or more tests compared to the profiles listed.

Alcaligenes↗
PubMed1979

[Isolation, purification and study of the stability of the soluble hydrogenase of Alcaligenes eutrophus Z-1].

Soluble hydrogenase was isolated from the hydrogen-oxidizing bacterium Alcaligenes eutrophus Z-1 and purified to electrophoretical homogeneity. The purification procedure included fractionation by ammonium sulfate, ion-exchange chromatography on DEAE-cellulose and gelfiltration through Ultragel AcA-34. The resulting preparation had a specific activity of 25 mkmoles H2.min-1.mg of protein as measured by the rate of hydrogen evolution from sodium dithionite-reduced methyl viologen. The enzyme has a molecular weight of 200,000 and is made up of two subunits with mol. weights of 30,000 and two subunits with mol. weights of 65,000. The effects of pH, oxidants and reducers, as well as aerobic and anaerobic conditions on the hydrogenase preparations inactivation kinetics in intact cells and in a highly purified state were studied. The kinetic data suggest a possible existence of two enzyme forms differing in their activities and stabilities to denaturating influences.

Air↗
PubMedDate not supplied

The efficacy of a temperature-sensitive mutant vaccine against Northwest Arkansas isolates of Alcaligenes faecalis.

The efficacy of a commercially produced temperature-sensitive mutant Alcaligenes faecalis vaccine was evaluated in turkeys contact-challenged with one of three strains of A. faecalis. In the vaccinated control group, the vaccine strain of A. faecalis colonized the nasal turbinates but not the trachea, caused no clinical signs of turkey coryza, and induced humoral antibodies. In the vaccinated challenged groups, the vaccine reduced both the severity of lesions and the number of birds exhibiting lesions compared with unvaccinated challenged groups, but it did not prevent colonization of challenge strains of A. faecalis.

Alcaligenes↗
PubMedDate not supplied

Alcaligenes faecalis infection in turkeys: effects on serum corticosterone and serum chemistry.

The serum levels of corticosterone, total calcium, inorganic phosphorus, protein, sodium, and potassium were determined for turkey poults from 1 to 42 days of age. Corticosterone levels were higher at days 10, 14, 21, and 28 in Alcaligenes faecalis-infected turkeys than in uninfected poults. The corticosterone responses in this study indicate that A. faecalis infection stimulates adrenal cortical function even in the very young poult, which ordinarily does not show adrenal cortical responsiveness to various environmental and physical stressors. No differences were observed between infected and uninfected turkey poults in other serum chemistries.

Alcaligenes↗
PubMed1985

Single and twinned crystals of ribulose-1,5-bisphosphate carboxylase-oxygenase from Alcaligenes eutrophus.

Ribulose-1,5-bisphosphate carboxylase-oxygenase (L8S8) from Alcaligenes eutrophus has been crystallized by equilibrium vapor diffusion techniques with ammonium sulfate as precipitant. Crystals thus obtained either as the ternary complex with CO2 and Mg2+ or as the quaternary complex with CO2, Mg2+, and 2-carboxyarabinitol 1,5-bisphosphate, a transition state analogue, diffract at least to 2.8-A resolution. Both are essentially isomorphous to each other, having orthorhombic space group C222(1) with cell dimensions a = 159 A, b = 159 A, and c = 200 A, and there is half a molecule in the asymmetric unit. The crystals of the ternary complex are sometimes twinned about the c axis so that the space group appears to be tetragonal. In this light, our earlier report (Bowien, B., Mayer, F., Spiess, E., Pähler, A., Englisch, U., and Saenger, W. (1982) Eur. J. Biochem. 106, 405-410) on a tetragonal space group P4(2)2(1)2 with crystals obtained from the same enzyme with Mg2+ and CO2 but without 2-carboxyarabinitol 1,5-bisphosphate might be incorrect.

Alcaligenes↗
PubMedDate not supplied

Partial characterization of the hemagglutinin of Alcaligenes faecalis.

The hemagglutinin of Alcaligenes faecalis was partially characterized. Hemagglutination (HA) was blocked by enzymes inactivating proteins, by heat, and by antisera but not by sugar-blocking substances. Pili were not determined to be a factor in HA activity. There was no connection between virulence and HA activity.

Alcaligenes↗
PubMed1978

Dependence on chain length of antitumor activity of (1 leads to 3)-beta-D-glucan from Alcaligenes faecalis var. myxogenes, IFO 13140, and its acid-degraded products.

The well-defined (1 leads to 3)-beta-D-glucan with (DPn) 540, produced by cultivation of Alcaligenes faecalis var. Myxogenes (IFO 13140), a mutant of a soil bacterium, had marked inhibitory activity against the s.c.-implanted Sarcoma 180 at 5 to 50 mg/kg for 10 days. It also exhibited very high activity in doses of 60 and 100 mg/kg i.p. by a single injection at 7 days after the initial s.c. transplantation of Sarcoma 180. The inhibition ratios observed with Ehrlich carcinoma, NTF (Nakahara-Tokuzen-Fukuoka) reticulum cell sarcoma, and CCM adenocarcinoma were somewhat less but were still significant. On the other hand the treatment failed to inhibit the growth of ascites Sarcoma 180 or to induce prolongation of life span. The mechanism of action of this glucan was considered to be host mediated because of a lack of effect in vitro and also because of the effectiveness of pretreatment of animals by injection before transplantation of a tumor. The results from the bioassay study of the low-molecular-weight (1 leads to 3)-beta-D-glucans prepared from the glucan with number-average degrees of polymerization (DPn) 540 by hydrolysis with formic acid or sulfuric acid showed that the glucans with DPn greater than 50 inhibited the growth of Sarcoma 180 implanted s.c. in mice, whereas the result of the schedule-dependent effect was obtained in the pretreatment of animals by the glucan with DPn 50. By i.v. administration, even the glucan with DPn 16 showed a strong antitumor effect comparable to that of the glucan with DPn 540.

Alcaligenes↗
PubMed1983

A cleavage product of mouse C3 adsorbable to (1----3)-beta-D-glucan from Alcaligenes faecalis var. myxogenes IFO 13140.

By using affinity chromatography of TAK, a gel-forming antitumor (1----3)-beta-D-glucan produced by Alcaligenes faecalis var. myxogenes IFO 13140 also known as an activator of the alternative complement pathway (ACP), we obtained from mouse serum a cleavage product of C3 (tentatively designated as C3X). Binding between C3X and TAK seemed to be noncovalent. In contrast to C3, C3X had an intact beta chain (70,000 daltons) and instead of an alpha chain, two peptide chains (40,000 and 30,000 daltons) linked by disulfide bonds. C3X retained a portion of C3 antigenicity. By trypsinolysis C3 yielded a fragment similar to C3X on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results suggest that C3X is C3c. Since C3X was obtained by elution with a high salt solution from TAK incubated with ethylenediaminetetraacetate (EDTA)-serum (serum containing EDTA) but not from TAK incubated with EDTA-plasma, C3X could already be present in the serum before the activation of ACP by TAK.

Adsorption↗
PubMedDate not supplied

Pathogenicity of various isolates of Alcaligenes faecalis for broilers.

Day-old broilers or specific-pathogen-free chickens were inoculated intranasally with approximately 1 X 10(8) organisms of eight different field isolates of Alcaligenes faecalis. Major differences in the pathogenicity of isolates and their ability to colonize the trachea were found. Only two isolates (Wilson and Lockamy) produced mild clinical signs of respiratory disease ("snicking," dyspnea). The same two also colonized the respiratory tract, especially the trachea, in large numbers; they persisted for 31 days. Of the remaining six isolates, five were also able to colonize the respiratory tract but did so to a lesser degree and less persistently, without causing clinical signs. Only one isolate (CS) was incapable of becoming established in the respiratory tract of chicks after intranasal inoculation.

Alcaligenes↗
PubMedDate not supplied

Observations on the pathogenicity of Alcaligenes faecalis in chickens.

A series of trials was conducted in which specific-pathogen-free (SPF) leghorn chicks were exposed to various isolates of Alcaligenes faecalis. Chicks were inoculated with A. faecalis alone or in combination with Newcastle disease/infectious bronchitis (Nc/Br) vaccine, laryngotracheitis vaccine, infectious bursal disease virus, or Mycoplasma gallisepticum. The response was evaluated by morbidity, mortality, airsacculitis, reisolation of A. faecalis, and histopathological lesions of tracheas. Although A. faecalis was recovered up to 42 days postinoculation in some cases, no clinical signs were directly attributed to simple A. faecalis infection. None of the other agents significantly increased the severity of A. faecalis signs or lesions, except that A. faecalis-infected chicks that were given Nc/Br vaccine had prolonged microscopic tracheal lesions. In another trial, the effects of A. faecalis in young SPF leghorns, non-SPF broilers, and turkeys were compared. Broiler-type chicks were more susceptible than leghorns and less susceptible than poults. Consequently, the use of leghorns as a model for studying this infection is questioned.

Alcaligenes↗
PubMedDate not supplied

Observations on Alcaligenes faecalis infection in turkeys.

Experiments were initiated to study the pathogenicity of 5 Alcaligenes faecalis isolates in specific-pathogen-free poults. The isolates were recovered from commercial flocks suffering from a respiratory disease. There were no differences between cultural or biochemical characteristics of the isolates, but differences in antibiotic sensitivity were detected. All 5 isolates were capable of initiating a respiratory disease in poults similar to that seen in the early stages of turkey coryza. The infection, clinical signs, and lesions were limited to the upper part of the respiratory tract, but there were substantial differences in the severity of disease initiated by different isolates. There were also differences in the persistence of infection in the host. Secondary infections in the tracheas and sinuses were higher in poults infected with A. faecalis. The disease observed in the experimentally infected birds was milder than in 4 naturally infected flocks that also had complicating Escherichia coli infections. There was no evidence of infection with infectious bursal disease virus in 4 naturally occurring outbreaks in Ohio. It is proposed that the term turkey coryza be used to describe the disease initiated by A. faecalis.

Alcaligenes↗
PubMed1984

Evidence for plasmid DNA in Alcaligenes faecalis.

A 10.6 megadalton plasmid was isolated from a virulent strain (NC-D) of Alcaligenes faecalis. Virulence and antibiotic sensitivity of this strain were compared with those characteristics of a mutant plasmid-free derivative, strain NC-D1. Strain NC-D1 was avirulent and lacked the streptomycin and sulfonamide resistances of the parent strain.

Alcaligenes↗
PubMedDate not supplied

Immediate hypersensitivity (anaphylaxis) response in uninfected and Alcaligenes faecalis-infected turkey poults.

The immediate hypersensitivity (anaphylaxis) response in uninfected and Alcaligenes faecalis-infected turkey poults was studied. The optimum challenge dose of bovine serum albumin (BSA) to elicit an anaphylactic response was 40 mg/kg body weight, and the optimum poult age was 4 to 5 weeks. The response latency time was 7 days. The anaphylactic response was specific for the sensitizing antigen used. No differences were observed between A. faecalis-infected and uninfected poults in the immediate hypersensitivity response to BSA.

Age Factors↗