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Following previous work in which a mass and monoxenous culture of Vorticella microstoma had been successfully established (Water Res., 7, 615 lpar;1973) another species of Ciliata, Colpidium campylum was subjected to continuous cultivation using Alcaligenes faecalis as the sole bacterial food and asparagine as the limiting substrate. This work was primarily undertaken to reveal the interaction and biological oscillation between these two types of organisms which simulate theecological behavior of activated sludge. The fact that the bacteria tended to flocculate and/ or deflocculate depending on the protozoan populastion density was incorporated into the rate equations to account for the oscillation in individual population density of the predator-prey system The mathematical approach presented earlier by canal and other workers forbiological oscillation used a homogeneously of the bacterial food wasoverelooked in the earlier publication.

More than 70 characters of 55 strains of Achromobacter xylosoxidans were compared with those of the type strain of A. xylosoxidans, ATCC 27061 (= KM 543). Repeated examination of these 55 strains confirmed the stability of the flagellar morphology and biochemical reaction pattern and performed that the species can be recognized by these characters. Two strains of Alcaligenes faecalis, two strains of Alcaligenes denitrificans, five strains of Alcaligenes sp., and five strains of each of King groups IIIa and IIIb were identified as strains of A. xylosoxidans. The base compositions of the deoxyribonucleic acids of 20 strains are given. The cellular fatty acid composition of extractable and bound lipids of five strains was determined. The minimal characters for the identification of strains of A. xylosoxidans are presented.

The descriptions of the genus Alcaligenes Castellani and Chalmers and Alcaligenes faecalis Castellani and Chalmers have been emended. It is requested that the Judicial Commission issue an Opinion rejecting the name Achromobacter Bergey et al. as a nomen dubium. It is proposed, on the basis of a comparison of type and reference strains, that Alcaligenes denitrificans Leifson and Hugh, Achromobacter arsenoxydans-tres Turner, and Alcaligenes odorans (MAlek and KazdovA-KožiskovA) MAlek et al. be considered as subjective synonyms of A. faecalis. The proposed taxonomic status of all known previously described species of the genera Alcaligenes and Achromobacter is indicated in appendixes.

Two electrophoretically distinguishable species of the 13S coupling factor of oxidative phosphorylation from Alcaligenes faecalis are detectable by standard polyacrylamide gel electrophoresis in the absence of urea, detergents, or any other protein-denaturing reagents. The slower species (type IA) can be converted into the faster species (type IB) by treatment with ATP, and the fast form converts into the slow form when aged at 4 degrees. The enzyme undergoes these conversions both when it is free in solution and when it is membrane bound. The ATP analog adenylyl imidodiphosphate (AMP-PNP) gives the conversion without being hydrolyzed and without causing any apparent change in the mass of the protein, which suggests that the conversion may be a ligand-induced conformational change. Types IA and IB can convert into three other electrophoretically distinguishable species (types IIA, IIB, and III) if the purification procedure involves chromatography on a DEAE-Sephadex column equilibrated in phosphate buffer. These conversions can be prevented if the column is eluted in morpholinoethanesulfonic acid (Mes) buffer and KCl. Type IIA is convertible into type IIB by ATP treatment. Types IA and IB will also convert into types IIA and IIB and finally into type III when aged for extended periods of time at 4 degrees, without a detectable change in mass. Coupling factor activity is lost when type I enzyme converts into type II enzyme, as is the ability of the enzyme to bind to the membrane. However, ATPase activity does not change significantly. The mitochondrial 13S coupling factor shows up to three electrophoretically distinguishable species. The use of phosphate buffer during DEAE-Sephadex chromatography gives conversion of slower species into faster species. ATP treatment does not give interconversions, and aging at 4 degrees gives only a slow dissociation of the enzyme into subunits. The chloroplast 13S coupling factor also shows up to three electrophoretic species. Incubation with ATP does not give interconversions, but a temperature-dependent conversion of the major species into a faster species occurs upon aging. The subunit composition of the three 13S enzymes is very similar by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, the major difference being in the number of classes of small polypeptides.

After the growth of Paecilomyces persicinus P-10 in a glucose-peptone medium, filtrates were collected and analyzed for antibiotic antivity. Activities against Salmonella gallinarum ATCC 3030 and Alcaligenes faecalis ATCC 8750 (penicillin N-resistant strain) were obtained. Part of the former activity was readily inactivated by penicillinase. The fraction active against A. faecalis was isolated by passage through Amberlite XAD-2 and Amberlite IRA-68. The powder eventually obtained was subjected to paper chromatography followed by bioautography, and the activity obtained corresponded to that of a sample of cephalosporin C. Thin-layer chromatography was also employed to verify the presence of cephalosporin C in the P-10 powder. The active solids were further purified by means of paper chromatography in a solvent system consisting of n -butanol-acetic acid-water (60:15:25, vol/vol). The material obtained from this procedure yielded an infrared absorption spectrum identical to that of cephalosporin C. Similarly, the ultraviolet absorption of the purified preparation coincided with that of cephalosporin C. Exposure of the purified solids to cephalosporinase resulted in rapid inactivation of the antibiotic. In addition to penicillin N and cephalosporin C, filtrates of P. persicinus P-10 also contained deacetylcephalosporin C, deacetoxycephalosporin C, and cephalosporin P.

RNAs extracted from crown-gall tumors, induced by Agrobacterium tumefaciens (strain B6) on stems of Datura stramonium L., have been isolated by the phenol method and purified through Biogel P 60 columns. These RNAs have been transformed into complexes with l-amino acids by incubation in a medium containing: Tris HCI buffer, pH 7.6, ATP, MgCl2, a mixture of l-amino acids and a polypeptide synthetase purified from Alcaligenes faecalis. The inoculation of stems of Datura plants with these complexes induces the development of nodular outgrowths, whereas other complexes made from RNAs isolated from healthy Datura plants, from Agrobacterium tumefaciens, Escherichia coli and Alcaligenes faecalis do not cause any hyperplasia under our experimental conditions. The analysis of the results obtained, supported by histological studies of the outgrowths, suggests that these neoformations should be of tumoral nature.

Taxonomical investigation was performed on the bacterium, strain NB 320 isolated from soil, and it was identified as Enterobacter cloacae. This bacterium produced the enzyme which catalyzed the transamination reaction between 3,4-dihydroxyphenyl pyruvate and an amino acid to form l-Dopa.The optimum culture conditions for the enzyme production were studied along with the characteristics of the enzyme. The enzyme of the strain was different in some properties from that of Alcaligenes faecalis IAM 1015 which had been already studied. The former utilized glutamate as an amino donor best among the amino acids tested for transamination and was induced by the addition of glutamine and asparagine. Intact cells of the strain did not catalyze the reaction unless they were treated with sonication or with a detergent.

An evaluation was made of media and tests used for differentiating nonfermenting gram-negative bacteria encountered in medical bacteriology in order to determine those diagnostic procedures most useful in identifying these bacteria. The organisms examined included Alcaligenes faecalis, A. odorans var. viridans, Moraxella duplex (Mima polymorpha var. oxidans), Acinetobacter anitratum (Herellea vaginicola), A. lwoffi (Mima polymorpha), Pseudomonas fluorescens, P. putida, P. maltophilia, P. pseudomallei, P. stutzeri, P. alcaligenes, and atypical strains of P. aeruginosa. The media and tests evaluated included Sellers' medium; Hugh and Leifson's OF medium; acid production from 10% lactose infusion agar; gluconate oxidation; starch, aesculin, and Tween 80 hydrolysis; lysine decarboxylase, arginine dihydrolase, deoxyribonuclease, and tyrosinase activity; tolerance to triphenyl tetrazolium chloride, cetrimide, cadmium sulfate, 2.5% and 6.5% sodium chloride, and p H 5.6; utilization of glucose, acetamide, and malonate.

KNOWLEDGE of the bacterial flora of the respiratory system of poultry artificially affected with aerosaccitis (air-sac infection) is incomplete. More information concerning organisms associated with avian PPLO (pleuropneumonia-like organisms) may be important in better understanding aerosaccitis and may lead to more effective methods of treatment. Price et al. (1957) isolated members of the genera Escherichia, Streptococcus, Staphylococcus, and Bacillus from the trachea of chickens with artificially induced aerosaccitis. Escherichia, pseudomonas, streptoccoci, staphylococci, bacilli, and corynebacteria were isolated from the pleural cavity (lungs and air sacs). Gram-negative organisms, mainly coliforms, were isolated more frequently from chickens with aerosaccitis than were gram-positive bacteria. Gram-positive organisms were isolated more frequently from normal birds. In addition, there was a great increase in the number of bacteria in the respiratory organs of infected birds. The purpose of this work was to investigate the nature and time of the changes in the respiratory flora of turkeys …

2,4-Dinitrophenol fails to stimulate the breakdown of the soluble high energy intermediate of oxidative phosphorylation isolated from Alcaligenes faecalis extracts. Once the intermediate is broken down, by other reactions, the coupling enzyme is prevented from reassociation with the electron transport particles by 2,4-dinitrophenol. Phosphorylating particles washed with 2,4-dinitrophenol lose coupling enzyme and with it the ability to form the energy-rich intermediate and to couple phosphorylation to electron transport. Phosphorylating particles containing bound coupling enzyme are able to form the high energy intermediate even in the presence of 2,4-dinitrophenol. However, after this first cycle of formation, no more can be formed because 2,4-dinitrophenol prevents the reassociation of coupling enzyme with the electron transport particles. 2,4-Dinitrophenol stimulates respiration in the bacterial phosphorylating particles.

Forty strains of Gram-negative rods which resembled Alcaligenes in failing to attack carbohydrates were studied. Twenty-four strains had lophotrichous flagella, three were monotrichous, three were either lophotrichous or monotrichous, six were peritrichous, and four were nonflagellated. Strains were grouped by their action on gelatin and nitrate, and by motility. Most lophotrichous and monotrichous organisms liquefied gelatin, reduced nitrate, or did both, whereas most peritrichous and nonflagellated rods failed to do either.Serological studies indicated antigenic heterogeneity, but some strains in the more homogeneous physiological groups were related, mainly through flagellar antigens. Three phages were obtained without induction but only one of these was lytic for a single heterologous strain. Inducible substances resembling bacteriocines were elaborated by several strains, and the activity was confined primarily to homogeneous physiological and serological groups. Some lophotrichous strains closely resembled but were not identical with the genus Lophomonas. The validity of the genus Alcaligenes is questioned, since few isolates resemble the original description (peritrichous flagella) and these could be included in the genus Achromobacter.

Formation of peptides from free amino acids by purified enzymes of Alcaligenes faecalis requires an RNA fraction. This RNA is capable of fixing all l -amino acids “activated” in the presence of each of the four ribonucleoside 5′-triphosphates and purified polypeptide synthetases. An “amino acid-RNA” complex was isolated. The acceptor fraction was identified as rapidly labelled RNA containing both “messenger” and eosomal RNA. Base analogs incorporated into this fraction modify its capacity for amino acids fixation.

SUMMARY: Several strains of the formerly described micro-organism then named Pseudomonas odorans Malek & Kazdova-Kožiskova were examined. It was found that this organism belongs to the genus Alcaligenes. A re-descript-of the species and a discussion of its taxonomic position is given.

A cellobiose-utilizing bacterium isolated from sugar cane bagasse and identified as a strain of Alcaligenes faecalis (ATCC 21400) produced an inducible β-glucoside-splitting enzyme. The enzyme was purified by a series of streptomycin and ammonium sulfate fractionations and by Sephadex and diethylaminoethyl column chromatography. The final preparation was purified 130-fold, with a recovery of about 10% of the initial enzyme activity. The enzyme had a wide p H range, with optimal activity at p H 6.0 to 7.0. The enzyme was stable in solution at p H 6.5 to 7.8 when kept at 30 C for 2 hr, but it was destroyed by temperatures above 55 C. At 58 and 60 C, the time required to inactivate 90% of the initial activity was 16 and 6.5 min, respectively. An activation energy of 9,500 cal/mole and a K m of 1.25 × 10 −4 m were obtained by using p -nitrophenyl β-glucoside as a substrate. The K i value and hydrolysis of cellobiose by the enzyme indicated a high affinity of the enzyme for the cellobiose. The enzyme had its specificity on β-glucosidic linkage and the rate of hydrolisis of glucosides depended upon the nature of the aglycon moiety. The inactivation studies showed the presence of sulfhydryl groups in the enzyme. The activity of the enzyme was easily destroyed by the Cu ++ and Hg ++ ions. The Michaelis-Menton relationship and the rate of heat inactivation indicated the presence of one type of noninteracting active site in the bacterial β-glucosidase. Molecular weight of the enzyme was estimated by gel filtration (Sephadex G-200) and sucrose density gradient, and a value of 120,000 to 160,000 was obtained.

As it is now possible as well as practical to identify nearly all nonfermentative Gram-negative bacilli recovered from clinical material, 565 strains of these bacteria were studied in respect to four features: incidence, distribution, potential pathogenicity, and antibiotic susceptibility. Four species, previously known to be opportunistic pathogens, Pseudomonas aeruginosa, P. maltophilia, Acinetobacter anitratus (Herellea vaginicola) , and A. Iwoffi (Mima polymorpha) , were the most commonly encountered. P. pseudoalcaligenes and P. fluorescens were also frequently encountered. Seven species of questionable pathogenicity were less common; these were P. stutzeri, P. acidovorans, P. putida, P. alcaligenes, P. multivorans, P. diminuta , and Chromobacterium typhiflavum. Bordetella bronchiseptica and Xanthomonas species, occasionally encountered, appeared to be nonpathogenic for man. Three of the five species of Moraxella associated with man were encountered: M. nonliquefaciens, M. osloensis , and M. phenylpyrouvica . Only one strain of Alcaligenes faecalis , of doubtful pathogenicity, was found. Some strains of A. odorans were pathogenic. Most strains of flavobacteria were not clinically significant; none of these corresponded biochemically to King's Flavobacterium meningosepticum .

The extracellular slime polysaccharide of Alcaligenes faecalis var. myxogenes strain 10C3 was isolated, purified, and shown to contain succinic acid (ca. 10%), glucose (ca. 70–80%), and small amounts of galactose and mannose. It seems to have β-glycosidic linkages. The polysaccharides obtained from cultures grown on glucose and on ethyleneglycol were studied. The two polymers had similar chemical and physical properties except that the polymer from glucose contained rather less mannose and had a higher specific viscosity than that from ethyleneglycol.

The microflora of healthy larvae and adults of the southern pine beetle, Dendroctonus frontalis , were determined by aseptically excising portions of the fore-, mid-, and hindguts and culturing them on agar. Aerobacter aerogenes, Alcaligenes faecalis , and Serratia marcescens were the principal species of bacteria recovered, and Aspergillus spp. and Penicillium spp. were the principal genera of fungi. Seven of the species recovered were identical with facultative or conditioned pathogens recovered from diseased beetles in other studies.

In the course of investigation of alicyclic hydrocarbon-utilizing microorganisms, five strains of ethylcyclohexane-utilizing bacteria were isolated from soil samples.Among those bacteria, the strain S6B1 that was identified as Alcaligenes faecalis, showed the best growth in shaking culture.The strain S6B1 was found to produce 4-ethylcyclohexanol from ethylcyclohexane.This substance separated from culture broth was purified and identified to be trans-4-ethylcyclohexanol by the use of NMR.

An attempt to show the effect of temperature on the rate of growth of bacteria in milk was made by finding the generation times ofSalmonella typhosa, Shigella dysenteriae, Streptococcus haemolyticus, Micrococcus pyogenes aureus, Lactobacillus acidophilus, Escherichia coli, Bacillus subtilisandAlcaligenes faecalisat temperatures ranging from 4 to 60°C. Growth was quantitatively measured by means of plate counts during the logarithmic period which was previously determined for each organism. The following is a summary of the results obtained:At 4° C. none of the micro-organisms showed evidence of multiplication during the 6hr.incubation.As temperatures of 5–45° C. were approached the generation time decreased until the optimum temperature for each organism was reached; beyond this point a slowing of growth was observed, until at 60° C. viability was apparently lost by all the organisms.Streptococcus haemolyticusandMicrococcus pyogenes aureushave a generation time of 37–23 min.Salmonella typhosaandShigella dysenteriaehave a generation time of 50–26 min.Escherichia coliandAlcaligenes faecalishave a generation time of 41–16 min.Bacillus subtilisgrows at about the same rate asAlcaligenes faecalis.The slowest organism of all appears to beLactobacillus acidophiluswith a generation time of 52–125 min.

The fine structure of curdlan produced by Alcaligenes faecalis var. myxogenes 10C3, mutant K, and pachyman from Poria cocos were investigated, with regard to their gel-forming properties. Periodate oxidation showed that both polysaccharides contain very high proportions of (1→3)-linked glucose residues. On complete hydrolysis the glucan polyalcohol obtained by periodate oxidation and borohydride reduction of curdlan (DP¯ 455) gave glucose and glycerol, in the molar ratio of 125~130: 1, and on mild acid hydrolysis yielded degraded polysaccharide (DP¯ 155), confirming the previous conclusion that curdlan has an essentially unbranched structure though it may contain a few internal (1→6)-glucosidic linkages. On complete hydrolysis the glucan polyalcohol derived from pachyman (DP¯ 255) gave glucose and glycerol, in the ratio of 40:1, and on mild hydrolysis it yielded degraded polysaccharide (DP¯ 130). This indicates that pachyman contains on the average four branch points and one internal (1→6)-glucosidic link...

1. Alcaligenes faecalis RN-4 was shown to have no detectable intra- or extracellular ribonuclease I (EC 2.7.7.16) activity. In this strain, ribosomal and messenger RNA's were degraded by polynucleotide phosphorylase (EC 2.7.7.8) and potassium-activated phosphodiesterase (EC 3.1.4.1) in vitro as demonstrated by the detection of nucleoside diphosphates and 5′-nucleotides as degradation products. 2. Mitomycin C induced the degradation of the ribosomal RNA of A. faecalis RN-4 in the same way as in Escherichia coli which is known to have a ribonuclease I. 3. In view of these results, the possibility that the ribonuclease I in Escherichia coli does not actually participate in the degradation of ribosomal RNA is discussed.

Interaction of β-chloro-l-[14C]alanine with l-aspartate-β-decarboxylase from Alcaligenes faecalis leads to inactivation associated with covalent binding of close to 1 mole of the 3-carbon chain of the analog per mole of active site. Evidence was obtained for two different types of binding. Thus, dialysis of the labeled enzyme against 0.05 m acetate (pH 6) led to loss of about half of the vitamin B6 cofactor without loss of 14C from the enzyme; before and after dialysis, about half of the 14C was released from the carboxyl group of the analog by treatment with ceric sulfate indicating the presence of a bound pyruvate residue. After dialysis of the labeled enzyme against 1 m acetate buffer (pH 5) containing 0.1 m l-glutamate, little additional vitamin B6 was lost (although similar dialysis of the holoenzyme releases all of the cofactor), but about half of the bound 14C was released; the 14C that remained bound to the dialyzed enzyme was not readily released by treatment with ceric sulfate. No evidence for nonidentical subunits was found. The data suggest that the two observed types of 14C-binding result from the aldimine-ketimine equilibrium; hydrolysis of the ketimine followed by loss of pyridoxamine 5'-phosphate at some sites competes with stabilization of the aldimine form by further interaction at other sites. The 14C-labeled enzyme was treated with cyanogen bromide and a labeled peptide was isolated. The label was released as β-hydroxy[14C]pyruvate from the peptide by treatment with mild alkali, 6 n HCl (105°, 18 hours), or on prolonged enzymatic digestion. Ammonolysis of the labeled peptide led to formation of glutamine and release of the label. The data indicate that the labeled derivative is β-hydroxypyruvate bound by ester linkage to a glutamic acid residue at the active center of the enzyme.

Lark, Cynthia (Saint Louis University, St. Louis, Mo.) and Robert Schichtel . Comparison of spheroplast induction in Alcaligenes faecalis by three different agents. J. Bacteriol. 84: 1241–1244. 1962.— Alcaligenes faecalis strain LB was exposed to different concentrations of cycloserine, d -methionine, or penicillin. The time course of spheroplast induction by these agents was measured as a function of their concentration. The results were consistent with models in which cycloserine reversibly inhibited an intracellular enzyme, and d -methionine was built into a defective cell-wall unit. Penicillin simulated cycloserine. This was taken as further evidence that penicillin inhibits an enzyme associated with cell-wall synthesis.

Oxidation of DPNH by phosphorylating particles from Alcaligenes faecalis is stimulated by P i and to a further variable degree by ADP plus P i . The rates of oxidation in the presence and absence of P i and ADP show different pH dependences. When a protein necessary for coupling phosphorylation to electron transport is removed from the particles, respiration is rapid and P i and ADP have no further effect. When this coupling protein (which is also found associated with DPN as an intermediate) is rebound to the particles by magnesium ions and a specific oligoribonucleotide, respiration is slowed, and can again be stimulated or released by P i or P i and ADP. The magnitude of respiratory stimulation by P i or P i and ADP is much smaller that that observed with intact mitochondria, but is similar to that observed with mitochondrial particles. These observations are compatible with our previously stated hypothesis that oxidative phosphorylation is mediated by electron carrier-coupling enzyme intermediates. The stimulating effect of P i or P i and ADP on respiration appears in this system to be the result of releasing electron carriers from such intermediates and thus allowing them to function again in electron transport.

Curdlan is produced by a mutant of Alcaligenes faecalis var. myxogenes, strain 10C3. The nature of its gel formation was investigated. The polymer formed a firm, resilient gel when heated in aqueous suspension at or above 54°C. An aqueous suspension (2%) of the polymer gave 730 (g/cm2) gel strength when heated at 90°C. The gel strength was independent of the incubation time but dependent upon the temperature. The presence of borate alone greatly increased the gel strength. The gel strength did not change between pH 2.5 and 10. The addition of urea, a reagent which breaks hydrogen bonds, caused a decrease in the gel-forming temperature, the extent of decrease depending upon the concentration of urea. X-ray studies indicated that heat-treatment of the polymer suspension caused a change in the molecular arrangement.

Succinoglucan, a succinylated polysaccharide produced by Alcaligenes faecalis var. myxogenes 10C3, was partially hydrolyzed with acid. Fractionation of the neutral oligosaccharides gave cellobiose, gentiobiose, laminaribiose, laminaritriose, 6-O-β-laminaribiosylglucose, 6-O-β-laminaritriosylglucose, and 3-O-β-cellobiosylgalactose, confirming the previous results that the polysaccharide consists of β-(l→3)-linked, (1→4)-linked and (1 →6)-linked d-glucose residues, and β-(1→3)-linked d-galactose residues.Possible structural features of succinoglucan were discussed on the basis of the above and previous results obtained by Smith degradation.

The enzymatic production of l-aspartic acid from maleic acid with cell suspensions of Alcaligenes faecalis 5-24, isolated from solid by the authors, was investigated.The optimum conditions of this reaction and some cultural conditions which influenced on the ability of the cells to catalyze the above reaction were mainly studied.The cells grown on maleic acid as a sole source of carbon showed exclusively the strong ability. The cells grown on a carbon source other than maleic acid showed no activity of this reaction.It was concluded that an inducibles enzyme whose formation was stimulated by the presence of maleic acid might be involved in the reaction for the production of l-aspartic acid from maleic acid.It was found that malonic acid was replaceable for maleic acid which played an inductive role for the formation of the enzyme system concerned with the reaction of l-aspartic acid production from maleic acid.The cells grown in the medium containing malonic acid showed a stronger activity of the above re...

A slimy non-spore-forming bacterium strain 10C3 isolated from soil was motile with peritrichous flagella and named Alcaligenes faecalis var. myxogenes. Studies were made on the conditions necessary for maximal production of a new acidic succinoglucan polysaccharide by this strain in shaken cultures. Much production was observed with sucrose, glucose, xylose, galactose, cellobiose, maltose, fructose, mannose and rhamnose. The yield was greatest with sucrose and decreased in order with the above sugars from about 36 to 23 per cent. The most suitable medium contained 4 per cent sugar, 0.5 per cent yeast extract and one per cent calcium carbonate in tap water. The optimum temperature was 28°C.