bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparative Study of 35 Bacteriophages of
Lactobacillus helveticus:
Morphology and Host Range
Authors: Séchaud Laurent , Rousseau Micheline , Fayard Blandine , Callegari Maria Luisa , Quénée Pascal , Accolas Jean-Pierre
Published: January 2020

This survey included 23 phages isolated from cheese whey and 12 temperate phages induced with mitomycin from their lysogenic host strains. All of the phages had an isometric head and a tail with a contractile sheath. In addition, short-tailed (160-nm-long) and long-tailed (260-nm-long) phages were distinguished. Short-tailed phages were by far the most widespread in French cheese factories (32 of the 35 phages studied). The study of phage relationships enabled two large groups of strains to be distinguished: those not or slightly sensitive to phages and those very sensitive to phages. There was an obvious relationship in the first group between phage sensitivity (or resistance) and the geographic origin of the strains. The second group contained primarily strains from large international collections and those isolated from commercial starters. The relationships among short-tailed phages, either temperate or isolated as lytic, suggest that lysogenic strains could be the major source of phages in French cheese factories.
DNA probe for Aeromonas salmonicida
Authors: Hiney M , Dawson M T , Heery D M , Smith P R , Gannon F , Powell R
Published: January 2020

A DNA fragment that is specific to Aeromonas salmonicida has been isolated from a genomic DNA library by differential hybridization. The specificity of this fragment as a DNA probe for A. salmonicida was shown by hybridization against reference strains and clinical isolates of A. salmonicida, related aeromonads, and species from several other bacterial genera. The sensitivity of detection by a polymerase chain reaction test, based on this fragment, was approximately two A. salmonicida cells.
Hydrophobicities and Electrophoretic Mobilities of Anaerobic Bacterial Isolates from Methanogenic Granular Sludge
Authors: Grotenhuis J. T. C. , Plugge C. M. , Stams A. J. M. , Zehnder A. J. B.
Published: January 2020

The hydrophobicities and electrophoretic mobilities of isolates from methanogenic anaerobic granular sludge were measured and compared with those of strains from culture collections. All new isolates were highly hydrophobic, indicating that the upflow anaerobic sludge blanket reactor concept selects for hydrophobic bacteria.
Methanothrix soehngenii
, a methanogen often observed in methanogenic granular sludge, was highly hydrophobic and showed low electrophoretic mobility at pH 7. The role of this strain in the formation of methanogenic granular sludge is discussed.
Comparison of extracellular peroxidase- and esterase-deficient mutants of Streptomyces viridosporus T7A
Authors: Magnuson T S , Crawford D L
Published: January 2020

Peroxidase-deficient mutants of the lignin-degrading bacterium Streptomyces viridosporus T7A were screened for their production of acid-precipitable polymeric lignin, extracellular peroxidases and esterases, and immunoreactivities against a polyclonal antibody produced against electrophoretically purified peroxidase isoform P3 of wild-type S. viridosporus. The mutants showed diminished abilities to solubilize lignin and produce acid-precipitable polymeric lignin. Their peroxidase activities were decreased, and their esterase production patterns were altered. Western immunoblots demonstrated that the mutants produced proteins immunologically reactive with the antibody, but with different mobilities from those of wild-type proteins. These findings confirm a direct role for peroxidases in lignin solubilization. They also indicate a possible role for esterases.
Metabolic Engineering To Produce Tyrosine or Phenylalanine in a Tryptophan-Producing
Corynebacterium glutamicum
Strain
Authors: Ikeda Masato , Katsumata Ryoichi
Published: January 2020

The aromatic amino acids are synthesized via a common biosynthetic pathway. A tryptophan-producing mutant of
Corynebacterium glutamicum
was genetically engineered to produce tyrosine or phenylalanine in abundance. To achieve this, three biosynthetic genes encoding the first enzyme in the common pathway, 3-deoxy-
d
-
arabino
-heptulosonate 7-phosphate synthase (DS), and the branch-point enzymes chorismate mutase and prephenate dehydratase were individually cloned from regulatory mutants of
C. glutamicum
which have either of the corresponding enzymes desensitized to end product inhibition. These cloned genes were assembled one after another onto a multicopy vector of
C. glutamicum
to yield two recombinant plasmids. One plasmid, designated pKY1, contains the DS and chorismate mutase genes, and the other, designated pKF1, contains all three biosynthetic genes. The enzymes specified by both plasmids were simultaneously overexpressed approximately sevenfold relative to the chromosomally encoded enzymes in a
C. glutamicum
strain. When transformed with pKY1 or pKF1, tryptophan-producing
C. glutamicum
KY10865, with the ability to produce 18 g of tryptophan per liter, was altered to produce a large amount of tyrosine (26 g/liter) or phenylalanine (28 g/liter), respectively, because the accelerated carbon flow through the common pathway was redirected to tyrosine or phenylalanine.
Genetic Analysis of the Role of Phytoalexin Detoxification in Virulence of the Fungus
Nectria haematococca
on Chickpea (
Cicer arietinum
)
Authors: Miao V. P. W. , Vanetten H. D.
Published: January 2020

Chickpea (
Cicer arietium
L.) produces the antimicrobial compounds (phytoalexins) medicarpin and maackiain in response to infection by microorganisms.
Nectria haematococca
mating population (MP) VI, a fungus pathogenic on chickpea, can metabolize maackiain and medicarpin to less toxic products. These reactions are thought to be detoxification mechanisms in
N. haematococca
MP VI and required for pathogenesis by this fungus on chickpea. In the present study, these hypotheses were tested by examining the phenotypes of progeny from crosses of the fungus that segregated for genes (
Mak
genes) controlling phytoalexin metabolism.
Mak1
and
Mak2
, two genes that individually confer the ability to convert maackiain to its 1a-hydroxydienone derivative, were linked to higher tolerance of the phytoalexins and high virulence on chickpea. These results indicate that this metabolic reaction is a mechanism for increased phytoalexin tolerance in the fungus, which thereby allows a higher virulence on chickpea.
Mak3
, a gene conferring the ability to convert maackiain to its 6a-hydroxypterocarpan derivative, also increased tolerance to maackiain in strains which carried it; however, the contribution of
Mak3
to the overall level of pathogenesis could not be evaluated because most progeny from the cross segregating for this gene were low in virulence. Thus, metabolic detoxification of phytoalexins appeared to be necessary, as demonstrated in the
Mak1
and
Mak2
crosses, but not sufficient by itself, as in the
Mak3
cross, for high virulence of
N. haematococca
MP VI on chickpea.
Seasonal effects on accumulation of microbial indicator organisms by Mercenaria mercenaria
Authors: Burkhardt W , Watkins W D , Rippey S R
Published: January 2020

The ability of hard-shelled clams (Mercenaria mercenaria) to accumulate fecal coliforms and other microorganisms (Escherichia coli, Clostridium perfringens, and male-specific bacteriophages) was determined over a 1-year period. Twenty separate trails were conducted during different seasons to encompass a wide range of water temperatures. The greatest accumulation of microorganisms in hard-shelled clams occurred during certain periods in the spring, at temperatures ranging from 11.5 to 21.5 degrees C. These periods of hyperaccumulation did not always coincide for all organisms; the accumulation of bacteriophages was not predicted by the accumulation of either fecal coliforms or C. perfringens. Bacteriophages and C. perfringens showed significantly higher rates of accumulation than either the fecal coliform group or E. coli, especially during the spring. The higher incidence of human viral gastroenteritis associated with the consumption of shellfish during this period may be a result of the extraordinary concentration of certain microorganisms, including enteric viral pathogens.
Model system using coliphage phi X174 for testing virus removal by air filters
Authors: Rapp M L , Thiel T , Arrowsmith R J
Published: January 2020

Short-term (15-min-duration) and long-term (5- to 6-day-duration) test procedures have been developed for determining the efficiency of the removal of bacteriophage phi X174 by air-sterilizing filters. These procedures were sensitive enough to measure a 10(8)-fold reduction in the number of bacteriophage. A filter commonly used in industrial air sterilizations (Domnick-Hunter Bio-X borosilicate glass) effected a 10(8)-fold removal of viable phage in both short-term and long-term tests. A prototype low-flux, hollow-fiber membrane gave similar results; however, a prototype high-flux, hollow-fiber membrane removed only about 99.999% of the bacteriophage in short-term tests.
Purification and characterization of an intracellular peroxidase from Streptomyces cyaneus
Authors: Mliki A , Zimmermann W
Published: January 2020

An intracellular peroxidase (EC 1.11.1.7) from Streptomyces cyaneus was purified to homogeneity. The enzyme had a molecular weight of 185,000 and was composed of two subunits of equal size. It had an isoelectric point of 6.1. The enzyme had a peroxidase activity toward o-dianisidine with a Km of 17.8 microM and a pH optimum of 5.0. It also showed catalase activity with a Km of 2.07 mM H2O2 and a pH optimum of 8.0. The purified enzyme did not catalyze C alpha-C beta bond cleavage of 1,3-dihydroxy-2-(2-methoxyphenoxy)-1-(4-ethoxy-3-methoxyphenyl) propane, a nonphenolic dimeric lignin model compound. The spectrum of the peroxidase showed a soret band at 405 nm, which disappeared after reduction with sodium dithionite, indicating that the enzyme is a hemoprotein. Testing the effects of various inhibitors on the enzyme activity showed that it is a bifunctional enzyme having catalase and peroxidase activities.
Energetics of arginine and lysine transport by whole cells and membrane vesicles of strain SR, a monensin-sensitive ruminal bacterium
Authors: Van Kessel J S , Russell J B
Published: January 2020

Strain SR, a monensin-sensitive, ammonia-producing ruminal bacterium, grew rapidly on arginine and lysine, but only if sodium was present. Arginine transport could be driven by either an electrical potential or a chemical gradient of sodium. Arginine was converted to ornithine, and it appeared that ornithine efflux created a sodium gradient which in turn drove arginine transport. There was a linear decline in arginine transport as pH was decreased from 7.5 to 5.5, and the cells did not grow at a pH less than 6.0. The Eadie-Hofstee plot was biphasic, and arginine could also be taken by a high-capacity diffusion mechanism. Because arginine was a strong inhibitor of lysine transport and lysine was a weak inhibitor of arginine transport, it appeared that both lysine and arginine were taken up by an arginine-lysine carrier which had a preference for arginine. The rate of lysine fermentation was always proportional to the extracellular lysine concentration, and facilitated diffusion was the dominant mechanism of lysine transport. When SR was grown in continuous culture on arginine or lysine, the theoretical maximal growth yield was similar (13 g of cells per mol of ATP), but the apparent maintenance energy requirement for arginine was greater than lysine (9.4 versus 4.4 mmol of ATP per g of cells per h). On the basis of differences in yield and maintenance energy, it appeared that active arginine transport accounted for approximately 40% of the total ATP.
Degradation of 1,2-dichloroethane by Ancylobacter aquaticus and other facultative methylotrophs
Authors: van den Wijngaard A J , van der Kamp K W , van der Ploeg J , Pries F , Kazemier B , Janssen D B
Published: January 2020

Cultures of the newly isolated bacterial strains AD20, AD25, and AD27, identified as strains of Ancylobacter aquaticus, were capable of growth on 1,2-dichloroethane (DCE) as the sole carbon and energy source. These strains, as well as two other new DCE utilizers, were facultative methylotrophs and were also able to grow on 2-chloroethanol, chloroacetate, and 2-chloropropionate. In all strains tested, DCE was degraded by initial hydrolytic dehalogenation to 2-chloroethanol, followed by oxidation by a phenazine methosulfate-dependent alcohol dehydrogenase and an NAD-dependent aldehyde dehydrogenase. The resulting chloroacetic acid was converted to glycolate by chloroacetate dehalogenase. The alcohol dehydrogenase was induced during growth on methanol or DCE in strain AD20, but no activity was found during growth on glucose. However, in strain AD25 the enzyme was synthesized to a higher level during growth on glucose than on methanol, and it reached levels of around 2 U/mg of protein in late-exponential-phase cultures growing on glucose. The haloalkane dehalogenase was constitutively produced in all strains tested, but strain AD25 synthesized the enzyme at a level of 30 to 40% of the total cellular protein, which is much higher than that found in other DCE degraders. The nucleotide sequences of the haloalkane dehalogenase (dhlA) genes of strains AD20 and AD25 were the same as the sequence of dhlA from Xanthobacter autotrophicus GJ10 and GJ11. Hybridization experiments showed that the dhlA genes of six different DCE utilizers were all located on an 8.3-kb EcoRI restriction fragment, indicating that the organisms may have obtained the dhlA gene by horizontal gene transmission.
Geographic distribution and genetics of killer phenotypes for the yeast Pichia kluyveri across the United States
Authors: Starmer W T , Ganter P F , Aberdeen V
Published: January 2020

Representative strains (n = 61) of the yeast Pichia kluyveri from across the United States were studied for their ability to kill 71 other strains (representing 25 species) of yeast. This survey showed killing activity in 69% of the P. kluyveri strains tested. More extensive analysis of killer activity of 197 P. kluyveri strains against strains of five tester species showed comparable activity (67% of strains tested). This activity was shown to be equally variable within localities, within regions, and across the continent. The genetic basis of the variability was ascertained by tetrad analysis and is most likely due to alleles segregating at three epistatic loci. Evidence for the idea that killer toxins have a role in excluding other yeasts from particular habitats is discussed.
Production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) in a recombinant Escherichia coli strain
Authors: Slater S , Gallaher T , Dennis D
Published: January 2020

An Escherichia coli strain has been constructed that produces the copolymer poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) P(HB-co-HV). This has been accomplished by placing the PHB biosynthetic genes from Alcaligenes eutrophus into an E. coli fadR atoC(Con) mutant and culturing the strain in M9 minimal medium containing glucose and propionate. 3-Hydroxyvalerate incorporation is absolutely dependent on the presence of both glucose and propionate, and 3-hydroxybutyrate-3-hydroxyvalerate ratios in the copolymer can be manipulated by altering the propionate concentration and/or the glucose concentration in the culture. P(HB-co-HV) production can be accomplished by using a wide variety of feeding regimens, but the most efficient is to allow the culture to grow to late log phase in minimal medium containing acetate and then add glucose and propionate to initiate copolymer production. A broad range of propionate concentrations can be used in the culture to stimulate 3-hydroxyvalerate incorporation; however, the most efficient utilization of propionate occurs at concentrations below 10 mM. 3-Hydroxyvalerate molar percentages in the copolymer are relatively constant over the course of growth. The copolymer has been purified and confirmed to be P(HB-co-HV) by gas chromatography/mass spectrometry and differential scanning calorimetry.
Effect of pH and Monensin on Glucose Transport by
Fibrobacter succinogenes
, a Cellulolytic Ruminal Bacterium
Authors: Chow Jo May , Russell James B.
Published: January 2020

Fibrobacter succinogenes
S85, a cellulolytic ruminal bacterium, required sodium for growth and glucose uptake. Cells which were deenergized with iodoacetate (500 μM) could not take up [
14
C]glucose. However, deenergized cells which were treated with valinomycin, loaded with potassium, and diluted into sodium or sodium plus potassium to create an artificial electrical gradient (ΔΨ) plus a chemical gradient of sodium (ΔpNa) or ΔpNa alone transported glucose at a rapid rate. Cells which were loaded with potassium plus sodium and diluted into sodium (ΔΨ with sodium, but no ΔpNa) also took up glucose at a rapid rate. Potassium-loaded cells that were diluted into buffers which did not contain sodium (ΔΨ without sodium) could not take up glucose. An artificial ZΔpH which was created by acetate diffusion could not drive glucose transport even if sodium was present. The maximum rate and affinity of glucose transport (pH 6.7) were 62.5 nmol/mg of protein per min and 0.51 mM, respectively. S85 was unable to grow at a pH of less than 5.5, and there was little glucose transport at this pH. When the extracellular pH was decreased, the glucose carrier was inhibited, intracellular pH declined, the cells were no longer able to metabolize glucose, and ΔΨ declined. Monensin (1 μM) or lasalocid (5 μM) decreased intracellular ATP and dissipated both the ΔΨ and ΔpNa. Since there was no driving force for transport, glucose transport was inhibited. These results indicated that
F. succinogenes
used a pH-sensitive sodium symport mechanism to take up glucose and that either a ΔΨ or a ΔpNa was required for glucose transport.
Degradation of hydrogen sulfide by Xanthomonas sp. strain DY44 isolated from peat
Authors: Cho K S , Hirai M , Shoda M
Published: January 2020

Xanthomonas sp. strain DY44, capable of degrading H2S, was isolated from dimethyl disulfide-acclimated peat. This bacterium removed H2S either as a single gas or in the presence of the sulfur-containing compounds methanethiol, dimethyl sulfide, and dimethyl disulfide. The maximum specific H2S removal rate, obtained in the late stationary phase, was 3.92 mmol g of dry cells-1 h-1 (6.7 x 10(-16) mol cell-1 h-1) at pH 7 and 30 degrees C through a batch experiment in a basal mineral medium. Since Xanthomonas sp. strain DY44 exhibited no autotrophic growth with H2S, the H2S removal was judged not to be a consequence of chemolithotrophic activity. By using X-ray photoelectron spectroscopy, the metabolic product of H2S oxidation was determined to be polysulfide, which has properties very similar to those of elemental sulfur. Autoclaved cells (120 degrees C, 20 min) did not show H2S degradation, but cells killed by gamma-irradiation and cell extracts both oxidized H2S, suggesting the existence of a heat-labile intracellular enzymatic system for H2S oxidation. When Xanthomonas sp. strain DY44 was inoculated into fibrous peat, this strain degraded H2S without lag time, suggesting that it will be a good candidate for maintaining high H2S removability in the treatment of exhaust gases.
Parameters Affecting Solvent Production by
Clostridium pasteurianum
Authors: Dabrock Birgit , Bahl Hubert , Gottschalk Gerhard
Published: January 2020

The effect of pH, growth rate, phosphate and iron limitation, carbon monoxide, and carbon source on product formation by
Clostridium pasteurianum
was determined. Under phosphate limitation, glucose was fermented almost exclusively to acetate and butyrate independently of the pH and growth rate. Iron limitation caused lactate production (38 mol/100 mol) from glucose in batch and continuous culture. At 15% (vol/vol) carbon monoxide in the atmosphere, glucose was fermented to ethanol (24 mol/100 mol), lactate (32 mol/100 mol), and butanol (36 mol/100 mol) in addition to the usual products, acetate (38 mol/100 mol) and butyrate (17 mol/100 mol). During glycerol fermentation, a completely different product pattern was found. In continuous culture under phosphate limitation, acetate and butyrate were produced only in trace amounts, whereas ethanol (30 mol/100 mol), butanol (18 mol/100 mol), and 1,3-propanediol (18 mol/100 mol) were the major products. Under iron limitation, the ratio of these products could be changed in favor of 1,3-propanediol (34 mol/100 mol). In addition, lactate was produced in significant amounts (25 mol/100 mol). The tolerance of
C. pasteurianum
to glycerol was remarkably high; growth was not inhibited by glycerol concentrations up to 17% (wt/vol). Increasing glycerol concentrations favored the production of 1,3-propanediol.
Polymerase chain reaction and gene probe detection of the 2,4-dichlorophenoxyacetic acid degradation plasmid, pJP4
Authors: Neilson J W , Josephson K L , Pillai S D , Pepper I L
Published: January 2020

Specific and sensitive detection of indigenous and introduced degradative organisms is an essential prerequisite to their use in remediation of toxic waste and soil systems. Procedures were employed for the use of polymerase chain reaction and gene probes for sensitive detection of the 2,4-dichlorophenoxyacetic-acid-degrading bacterium, Alcaligenes eutrophus JMP134(pJP4). Two 20-mer oligonucleotide primers were identified for amplification of a 205-bp region of the tfdB gene of pJP4, and optimum conditions for amplification were determined. Both the polymerase chain reaction amplification process and hybridization with the 5'-end-labelled probe were found to be specific to organisms containing plasmid pJP4 or its derivative pRO103. Detection limits were determined for the template supplied either as bacterial cells or purified plasmid DNA. The detection was sensitive up to an initial inoculum of 3,000 CFU or 156 pg of total plasmid DNA. However, when the amplified product was transferred to a nylon membrane and hybridized with the 5'-end-labelled probe, the detection sensitivity increased to 300 CFU or 15.6 pg of plasmid DNA. This sensitive detection method is more specific than use of traditional indicator media (M. A. Loos, Can. J. Microbiol. 21:104-107, 1975). An oligonucleotide (20 bases) complementary to a sequence internal to the 205-bp region was synthesized and utilized as a probe to confirm the specificity of the detection.
Relationship between Desiccation and Exopolysaccharide Production in a Soil
Pseudomonas
sp
Authors: Roberson Emily B. , Firestone Mary K.
Published: January 2020

The relationship between desiccation and the production of extracellular polysaccharides (EPS) by soil bacteria was investigated by using a
Pseudomonas
species isolated from soil. Cultures subjected to desiccation while growing in a sand matrix contained more EPS and less protein than those growing at high water potential, suggesting that resources were allocated to EPS production in response to desiccation. Desiccation did not have a significant effect on activity as measured by reduction of iodonitrotetrazolium. Purified EPS produced by the
Pseudomonas
culture contained several times its weight in water at low water potential. Sand amended with EPS held significantly more water and dried significantly more slowly than unamended sand, implying that an EPS matrix may buffer bacterial colonies from some effects of desiccation. We conclude that bacteria may use EPS production to alter their microenvironment to enhance survival of desiccation.
Growth Characteristics of the Thermophilic Fungus
Scytalidium thermophilum
in Relation to Production of Mushroom Compost
Authors: Wiegant W. M.
Published: January 2020

Scytalidium thermophilum
is an important thermophilic fungus in the production of mushroom compost. I investigated the characteristics of this organism and present a simple model with which fungal growth in compost can be described. The model is used to predict better circumstances for rapid indoor production of mushroom compost. I conclude that inoculation of the starting material with prepared compost either before or after the pasteurization phase has only a minor effect on the shortening of the composting process. This is because the initial growth rate of the fungus is much higher than its growth rate later. A lower temperature (53.5°C instead of the usual 56 to 58°C) during the pasteurization phase may be most profitable for rapid compost production; such a temperature may reduce the time that is needed for the last phase of the production process by at least 1 day.
Evidence for an NIH shift in oxidation of naphthalene by the marine cyanobacterium Oscillatoria sp. strain JCM
Authors: Narro M L , Cerniglia C E , Van Baalen C , Gibson D T
Published: January 2020

The marine cyanobacterium Oscillatoria sp. strain JCM oxidized naphthalene predominantly to 1-naphthol. Experiments with [1-2H]naphthalene and [2-2H]naphthalene indicated that 1-naphthol was formed with 68 and 74% retention of deuterium, respectively. No significant isotope effect was observed when the organism was incubated with a 1:1 mixture of naphthalene and [2H8]naphthalene. The results indicate that 1-naphthol is formed through a naphthalene 1,2-oxide intermediate, which rearranges spontaneously via an NIH shift mechanism.
Metabolism of Phosphonoacetate as the Sole Carbon and Phosphorus Source by an Environmental Bacterial Isolate
Authors: McMullan G. , Harrington F. , Quinn J. P.
Published: January 2020

A gram-negative bacterium isolated from activated sludge was able to utilize up to 25 mM phosphonoacetate as the sole carbon and phosphorus source, with simultaneous excretion of virtually equimolar levels of phosphate. 2-Aminoethylphosphonate was similarly utilized with equivalent growth rates and cellular yields, while 3-aminopropyl-, 4-aminobutyl-, methyl-, ethyl-, and phenylphosphonates served only as phosphorus sources.
Bacterial Hydrolysis of Protein and Methylated Protein and Its Implications for Studies of Protein Degradation in Aquatic Systems
Authors: Keil Richard G. , Kirchman David L.
Published: January 2020

Ribulose 1,5-bisphosphate carboxylase was radiolabelled by in vitro translation, resulting in uniformly labelled ribulose 1,5-bisphosphate carboxylase, and also by reductive methylation. We investigated the degradation of the two forms of radiolabelled protein by natural bacterial populations. Although total hydrolysis of uniformly labelled protein and methylated protein was nearly equal, percent assimilation, respiration, and release as low-molecular-weight material were different. Radioactivity from uniformly labelled protein was approximately equally assimilated into cells, respired as
3
H
2
O, and released as low-molecular-weight material, but radioactivity from the methylated protein was nearly all released as low-molecular-weight material, and little was assimilated or respired.
Method to identify specific alleles of a Phanerochaete chrysosporium gene encoding lignin peroxidase
Authors: Gaskell J , Vanden Wymelenberg A , Stewart P , Cullen D
Published: January 2020

A method to identify and differentiate allelic variants of the gene encoding lignin peroxidase isozyme H8 is presented. The strategy involves amplifying a variable region of the gene's carboxy terminus by use of the polymerase chain reaction and then probing with allele-specific oligonucleotides.
Stimulation of Reductive Dechlorination of Tetrachloroethene in Anaerobic Aquifer Microcosms by Addition of Short-Chain Organic Acids or Alcohols
Authors: Gibson Susan A. , Sewell Guy W.
Published: January 2020

The effect of the addition of common fermentation products on the dehalogenation of tetrachloroethene was studied in methanogenic slurries made with aquifer solids. Lactate, propionate, crotonate, butyrate, and ethanol stimulated dehalogenation activity, while acetate, methanol, and isopropanol did not.
Factors Stimulating Propagation of Legionellae in Cooling Tower Water
Authors: Yamamoto Hiroyuki , Sugiura Minoru , Kusunoki Shinji , Ezaki Takayuki , Ikedo Masanari , Yabuuchi Eiko
Published: January 2020

Our survey of cooling tower water demonstrated that the highest density of legionellae, ≥10
4
CFU/100 ml, appeared in water containing protozoa, ≥10
2
MPN/100 ml, and heterotrophic bacteria, ≥10
6
CFU/100 ml, at water temperatures between 25 and 35°C. Viable counts of legionellae were detected even in the winter samples, and propagation, up to 10
5
CFU/100 ml, occurs in summer. The counts of legionellae correlated positively with increases in water temperature, pH, and protozoan counts, but not with heterotrophic bacterial counts. The water temperature of cooling towers may promote increases in the viable counts of legionellae, and certain microbes, e.g., protozoa or some heterotrophic bacteria, may be a factor stimulating the propagation of legionellae.
Modelling of mixed chemostat cultures of an aerobic bacterium, Comamonas testosteroni, and an anaerobic bacterium, Veillonella alcalescens: comparison with experimental data
Authors: Gerritse J , Schut F , Gottschal J C
Published: January 2020

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 individual species in (oxygen-limited) chemostats. Using these pure-culture data in the model of the mixed culture resulted in a good description of the actual mixed cultures of the two bacteria. In the actual mixed-culture experiments, coexistence of the two species occurred only when the cultures were oxygen limited. With increasing oxygen supply (the actual oxygen concentration in the culture remaining at less than 0.2 microM), the biomass of C. testosteroni increased, whereas that of V. alcalescens decreased. Apparently, C. testosteroni protected V. alcalescens from inhibition by oxygen by maintaining sufficiently low oxygen concentrations. The model calculations indicated that competition between the aerobic and the anaerobic bacterium for common substrates (L-lactate and oxygen) occurred and that the anaerobe was the better competitor. Analysis of the culture fluid indicated that C. testosteroni grew primarily at the expense of the fermentation products of V. alcalescens, i.e., propionate and acetate. The model further indicated that with different values of several growth parameters (e.g., substrate affinity and/or inhibition constants), the affinity of the aerobic organism for oxygen and the sensitivity of the anaerobic organism for oxygen were the most important properties determining the coexistence of these two physiologically different types of bacteria.
Identification and cloning of Bradyrhizobium japonicum genes expressed strain selectively in soil and rhizosphere
Authors: Bhagwat A A , Keister D L
Published: January 2020

The growth of Bradyrhizobium japonicum USDA 110 and USDA 438 in soil extract-supplemented medium led to transcription of a large amount of DNA not expressed in basal medium. Strain USDA 438 was more competitive for the nodulation of soybean than strain USDA 110. To identify and isolate DNA regions which were expressed specifically in strain USDA 438 but not in strain USDA 110 in response to soil extract or soybean root exudate, we developed a subtractive RNA hybridization procedure. Several cosmid clones which showed strain-specific gene expression were isolated from a USDA 438 gene library. Two clones enhanced competitive nodulation when mobilized to USDA 110. The method described may be useful for identifying genes expressed in response to environmental stimuli or genes expressed differently in related microbial strains.
Cloning and partial characterization of two chromosomal loci from Bacteroides ovatus that contain genes essential for growth on guar gum
Authors: Valentine P J , Arnold P , Salyers A A
Published: January 2020

Previously, we isolated three transposon insertion mutants of Bacteroides ovatus (M-4, M-5, and M-7) that were unable to grow on the branched polysaccharide guar gum. In this study, we used a tetracycline resistance gene on the transposon to clone chromosomal DNA adjacent to the transposon insertions in each of the three mutants. Restriction analysis of the flanking chromosomal DNA in M-4 and M-7 revealed that the insertions in these two mutants were in the same location. The cloned DNA adjacent to the insertions in M-5 and M-7 was used as a hybridization probe to clone the wild-type loci. Two clones of about 10 kbp in size were obtained. Restriction analysis showed that these two clones did not overlap. The clone of the M-5 locus appeared to contain all of the genes affected by the M-5 insertion, but we were unable to demonstrate complementation of the M-5 mutation because of the instability of the clone in this background. Analysis of the clone of the M-7 locus showed that it contained a guar gum-regulated promoter, but the transcript originating from this promoter was not affected by the transposon insertion. Thus, the M-7 locus apparently contains at least two separate transcriptional units, the one defined by this promoter and the one interrupted by the transposon insertion. Insertion mutations downstream of the guar gum-regulated promoter demonstrated that there were essential guar gum utilization genes in this region. The M-7 mutant was eliminated by the wild type in the intestinal tracts of germfree mice.(ABSTRACT TRUNCATED AT 250 WORDS)
Comparison of contact angles and adhesion to hexadecane of urogenital, dairy, and poultry lactobacilli: effect of serial culture passages
Authors: Reid G , Cuperus P L , Bruce A W , van der Mei H C , Tomeczek L , Khoury A H , Busscher H J
Published: January 2020

The aim of this study was to examine the hydrophobicities of 23 urogenital, dairy, poultry, and American Type Culture Collection isolates of lactobacilli and to determine the effect on hydrophobicity of serially passaging the strains in liquid medium. To this end, strains were grown after isolation and identification and then serially passaged up to 20 times. Hydrophobicity was assessed through contact angle measurements on lawns of cells by using water, formamide, methylene iodide, 1-bromonaphthalene, and hexadecane as wetting agents and through measurement of their partitioning in a hexadecane-water system. The hydrophobicities of these strains varied widely, with Lactobacillus casei strains being predominantly hydrophilic and L. acidophilus strains being mostly hydrophobic. For some isolates, serial passaging was accompanied by a clear loss of hydrophobic surface properties, whereas for other strains, cultures became heterogeneous in that some cells had already lost their hydrophobic surface properties while others were still hydrophobic. Adhesion of this collection of lactobacilli to hexadecane droplets in microbial adhesion to hexadecane (MATH) tests was driven by their aversion to water rather than by their affinity for hexadecane, as concluded from the fact that hexadecane contact angles were zero for all strains. Furthermore, adhesion of the lactobacilli to hexadecane in MATH tests occurred only when the water contact angle on the cells was above 60 degrees.
Mineralization of glucose and lignocellulose by four arctic freshwater sediments in response to nutrient enrichment
Authors: McKinley V L , Vestal J R
Published: January 2020

Microbial biomass and activity were examined in four different arctic sediments: littoral lake sediment and profundal lake sediment from Toolik Lake, Alaska, thaw pond sediment, and eroding river bank peat. The thaw pond sediment had the largest viable microbial biomass, while the profundal sediment had the smallest. Rates of glucose or acetate incorporation into lipids, glucose mineralization, and lignocellulose mineralization (all normalized per unit of biomass) were highest in the river peat sample, however. The kinetics of glucose mineralization in the profundal sediment were very different from those in the other three samples: although the initial rate of mineralization was five times lower than that in the peat and two times lower than that in the littoral and thaw pond sediments, the maximum amount of 14CO2 evolved from [14C]glucose eventually equaled that in the peat and exceeded that in the littoral and thaw pond sediments by 2.0 and 3.5 times, respectively. Carex aquatilis [14C-cellulose]- and [14C-lignin]lignocellulose mineralization rates in the profundal sediment equaled or exceeded those in the littoral sediment after 16 and 46 days, but the pattern of nutrient limitation differed: the profundal sediment was the only one sampled that exhibited nitrogen limitation, while the other three sediments appeared to be limited primarily by phosphorus. The addition of nitrogen and phosphorus together had no cumulative effects on lignocellulose mineralization. When the rates of mineralization or incorporation of glucose are compared with those of lignocellulose, the results of this study indicate that profundal sediment communities may be better able to utilize the more recalcitrant substrates relative to the labile substrates than microbial communities from sediments rich in detritus and standing macrophytes.
Evidence that some Frankia sp. strains are able to cross boundaries between Alnus and Elaeagnus host specificity groups
Authors: Bosco M , Fernandez M P , Simonet P , Materassi R , Normand P
Published: January 2020

Phenotypic and genotypic methods were used to prove the existence of Frankia strains isolated from an Elaeagnus sp. that are able to cross the inoculation barriers and infect Alnus spp. also. Repeated cycles of inoculation, nodulation, and reisolation were performed under axenic conditions. Frankia wild-type strain UFI 13270257 and three of its coisolates did exhibit complete infectivity and effectiveness on Elaeagnus spp. and Hippophaë rhamnoides and variable infectivity on Alnus spp. Microscopical observation of host plant roots showed that these strains are able to infect Alnus spp. by penetrating deformed root hairs. Reisolates obtained from nodules induced on monoxenic Alnus glutinosa, Alnus incana, and Elaeagnus angustifolia resembled the parent strains in host infectivity range, in planta and in vitro morphophysiology, isoenzymes, and nif and rrn restriction fragment length polymorphisms, thus fulfilling Koch's postulates on both host plant genera. Alnus and Elaeagnus group-specific polymerase chain reaction DNA amplifications, DNA-DNA hybridizations, and partial gene sequences coding for 16S rRNA provided evidence for the genetic uniformity of wild-type strains and their inclusion into one and the same genomic species, clearly belonging to the Elaeagnus group of Frankia species.
Styrene formation by the decomposition by Pichia carsonii of trans-cinnamic acid added to a ground fish product
Authors: Shimada K , Kimura E , Yasui Y , Tanaka H , Matsushita S , Hagihara H , Nagakura M , Kawahisa M
Published: January 2020

It is not well known how the formation of styrene by microorganisms can occur in foods. In this study, we described and characterized the production of styrene by a yeast isolated from chikuwa fish paste. The styrene was not detected in fresh and normal food products nor in the food package's plastic film. The food containing styrene contained cinnamic acid as an antimicrobial agent and spice, and it was contaminated by 5.4 x 10(6) CFU of a yeast per gram. On the basis of morphological and biochemical features, the yeast isolated was determined to be a strain of Pichia carsonii, now designated strain CHI. Strain CHI, which was able to grow on cinnamic acid, had the ability to form styrene from trans-cinnamic acid via trans-p-coumaric and caffeic acids. The MIC of trans-cinnamic acid against strain CHI was 230 micrograms/ml. Strain CHI thrived well at pH 5.0 and 26.0 degrees C and was tolerant to 20% NaCl. Styrene was subsequently produced in ground fish meat containing cinnamic acid into which strain CHI had been inoculated. The yeast was found to be an environmental contaminant in food processing plants of the chikuwa manufacturer.
Some growth and metabolic characteristics of monensin-sensitive and monensin-resistant strains of Prevotella (Bacteroides) ruminicola
Authors: Morehead M C , Dawson K A
Published: January 2020

New strains with enhanced resistance to monensin were developed from Prevotella (Bacteroides) ruminicola subsp. ruminicola 23 and P. ruminicola subsp. brevis GA33 by stepwise exposure to increasing concentrations of monensin. The resulting resistant strains (23MR2 and GA33MR) could initiate growth in concentrations of monensin which were 4 to 40 times greater than those which inhibited the parental strains. Resistant strains also showed enhanced resistance to nigericin and combinations of monensin and nigericin but retained sensitivity to lasalocid. Glucose utilization in cultures of the monensin-sensitive strains (23 and GA33) and one monensin-resistant strain (23MR2) was retarded but not completely inhibited when logarithmic cultures were challenged with monensin (10 mg/liter). Monensin challenge of cultures of the two monensin-sensitive strains (23 and GA33) was characterized by 78 and 51% decreases in protein yield (milligrams of protein per mole of glucose utilized), respectively. Protein yields in cultures of resistant strain 23MR2 were decreased by only 21% following monensin challenge. Cell yields and rates of glucose utilization by resistant strains GA33MR were not decreased by challenge with 10 mg of monensin per liter. Resistant strains produced greater relative proportions of propionate and less acetate than the corresponding sensitive strains. The relative amounts of succinate produced were greater in cultures of strains 23, GA33, and 23MR2 following monensin challenge. However, only minor changes in end product formation were associate with monensin challenge of resistant strain GA33MR. These results suggest that monensin has significant effects on both the growth characteristics and metabolic activities of these predominant, gram-negative ruminal bacteria.
Overproduction of threonine by Saccharomyces cerevisiae mutants resistant to hydroxynorvaline
Authors: Ramos C , Calderon I L
Published: January 2020

In this work, we isolated and characterized mutants that overproduce threonine from Saccharomyces cerevisiae. The mutants were selected for resistance to the threonine analog alpha-amino-beta-hydroxynorvalerate (hydroxynorvaline), and, of these, the ones able to excrete threonine to the medium were chosen. The mutant strains produce between 15 and 30 times more threonine than the wild type does, and, to a lesser degree, they also accumulate isoleucine. Genetic and biochemical studies have revealed that the threonine overproduction is, in all cases studied, associated with the presence in the strain of a HOM3 allele coding for a mutant aspartate kinase that is totally or partially insensitive to feedback inhibition by threonine. This enzyme seems, therefore, to be crucial in the regulation of threonine biosynthesis in S. cerevisiae. The results obtained suggest that this strategy could be efficiently applied to the isolation of threonine-overproducing strains of yeasts other than S. cerevisiae, even those used industrially.
Acyloin formation by benzoylformate decarboxylase from Pseudomonas putida
Authors: Wilcocks R , Ward O P , Collins S , Dewdney N J , Hong Y , Prosen E
Published: January 2020

Whole cells and cell extracts of Pseudomonas putida grown in a medium containing ammonium mandelate have the capacity to produce the acyloin compound 2-hydroxypropiophenone when incubated with benzoylformate and acetaldehyde. Benzaldehyde and benzyl alcohol were formed as reaction by-products. The enantiomeric excess of the 2-hydroxypropiophenone product was found to be 91 to 92%. The absolute configuration of the enzymatically prepared product at the carbinol carbon was found to be S. The thiamine PPi-linked enzyme benzoylformate decarboxylase, purified to give a single protein band on polyacrylamide gel electrophoresis, was shown to be responsible for the catalysis of this novel condensation reaction.
Determination of the respiration kinetics for mycelial pellets of Phanerochaete chrysosporium
Authors: Michel F C , Grulke E A , Reddy C A
Published: January 2020

In mycelial pellet cultures of the white rot basidiomycete Phanerochaete chrysosporium, low oxygen concentration negatively affects the production of the extracellular lignin peroxidases and manganese peroxidases which are key components of the lignin-degrading system of this organism. To test the hypothesis that oxygen limitation in the pellets is responsible for this effect, oxygen microelectrodes were used to determine oxygen concentration gradients within the mycelial pellets of P. chrysosporium. Pellets were removed from oxygenated cultures, allowed to equilibrate with air, and probed with oxygen microelectrodes. The oxygen profiles were modelled assuming that O2 uptake follows a Michaelis-Menten relationship. The Vmax and Km values for oxygen uptake were 0.76 +/- 0.10 g/m3 of pellet per s and 0.5 +/- 0.3 g/m3, respectively. These kinetic values were used to predict respiration rates in air-flushed cultures, oxygen-flushed cultures, and cultures with large pellets (diameter greater than 6 mm). The predicted respiration rates were independently validated by experimentally measuring the evolution of carbon dioxide from whole cultures.
Pseudomonas cepacia suppression of sunflower wilt fungus and role of antifungal compounds in controlling the disease
Authors: McLoughlin T J , Quinn J P , Bettermann A , Bookland R
Published: January 2020

In a field experiment, Pseudomonas cepacia J82rif and J51rif increased sunflower emergence in the presence of the fungus Sclerotinia sclerotiorum. Pyrrolnitrin, aminopyrrolnitrin, and monochloroaminopyrrolnitrin were isolated from J82 and identified by using thin-layer chromatography, high-performance liquid chromatography, and electron impact-mass, UV, and infrared spectroscopy. In growth chamber experiments, two antibiosis-negative mutants were not different from the parent strain in protecting the seeds from the fungus.
Preparation of Refractile Spores of
Clostridium thermosaccharolyticum
Involves a Solventogenic Phase
Authors: Landuyt Sandra L. , Hsu Edward J.
Published: January 2020

Conversion of vegetative cells of
Clostridium thermosaccharolyticum
to refractile endospores was achieved by sequential transfer and dilution at each generation, with a final dilution into a sporulation medium that contained xylan supplemented with excess calcium. The subsequent growth was synchronous and resulted in elongated, solventogenic cells that were then shifted to 35°C to permit further differentiation without cell division. The synchronized cells grown in xylan medium supplemented with Ca gluconate produced total solvents that reached 9.63% (vol/vol). One hundred percent of these elongated solventogenic cells (4.84 × 10
9
cells per ml) entered the sporangial stage and continued to differentiate into refractile spores. Only cells sequentially transferred and diluted at a critical time of the growth cycle are synchronized, induced to elongate (≥fourfold), become highly solventogenic in the presence of excess calcium, and are converted to a homogeneous population of refractile spores.