bluuebunny/crossref_metadata_2025 huggingface dataset:
Carbamyl Phosphate and Acetyl Phosphate Synthesis in
Escherichia coli:
Analysis of Associated Enzyme Activities by an Antibody to Acetokinase
Authors: Brzozowski Thomas H. , Kalman Sumner M.
Published: January 2020

Brzozowski, Thomas
H. (Stanford University School of Medicine, Palo Alto, Calif.),
and Sumner M. Kalman
. Carbamyl phosphate and acetyl phosphate synthesis in
Escherichia coli
: analysis of associated enzyme activities by an antibody to acetokinase. J. Bacteriol.
91:
2286–2290. 1966.—Earlier studies have shown that the carbamyl phosphate synthesis from ammonia in cell extracts of wild-type
Escherichia coli
is due to at least two enzymes, acetokinase and the glutamine-dependent carbamyl phosphate synthetase. Partial purification of the glutamine-dependent carbamyl phosphate synthetase and acetokinase fails to separate from these enzymes this ammonia-dependent activity. An antibody to the partially purified acetokinase was prepared and used to determine the distribution of the ammonia-dependent activity in wild-type organisms and single-step arginine-uracil-requiring mutants with respect to the two enzymes. Such a study was possible because the antibody inhibits acetokinase but not the glutamine-utilizing carbamyl phosphate synthetase. Enzyme inhibition obtained by the stepwise addition of the antibody to cell extracts indicates that all of the ammonia-dependent carbamyl phosphate synthesis observed in the arginine-uracil-requiring mutants is due to a protein in the acetokinase fraction, presumably acetokinase itself, since acetyl phosphate and carbamyl phosphate synthesis were inhibited in a parallel fashion. In wild-type organisms, there is only partial inhibition of the ammonia-dependent activity, even when enough antibody is added to produce maximal inhibition of acetokinase. It is suggested that this residue is due to the glutamine-dependent carbamyl phosphate synthetase, for the ratio of the antibody insensitive to antibody sensitive ammonia-dependent activity present in cell extracts of the two wild-type organisms reported is qualitatively proportional to the level of carbamyl phosphate synthetase present relative to acetokinase.
Availability of Bases and Nucleosides as Precursors of Nucleic Acids in L Cells and in the Agent of Meningopneumonitis
Authors: Tribby Ilse I. E. , Moulder James W.
Published: January 2020

Tribby, IlseI. E. (University of Chicago, Chicago, Ill.),and James W. Moulder. Availability of bases and nucleosides as precursors of nucleic acids in L cells and in the agent of meningopneumonitis. J. Bacteriol.91:2362–2367. 1966.—Uninfected L cells and the meningopneumonitis agent propagated in L cells utilized exogenous adenine, guanine, and their ribonucleosides and deoxyribonucleosides for synthesis of both deoxyribonucleic acid (DNA) and ribonucleic acid. Cytosine, cytidine, and uridine were also incorporated into the nucleic acids of both host and parasite. L cells and the meningopneumonitis agent incorporated uracil, thymine, and deoxyuridine very poorly. L cells utilized thymidine and deoxycytidine almost exclusively for DNA synthesis, but the meningopneumonitis agent did not incorporate these nucleosides at all. Since the L cell had previously been shown to convert added thymidine to its nucleotides, mainly the triphosphate, it was concluded that the meningopneumonitis agent can utilize neither the thymidine nor the thymidine nucleotides of the L-cell pool, and that it can probably synthesize the thymidine triphosphate needed for DNA synthesis from the uridine of the L-cell pool.
Effect of Amino Acids and Oxygen on Chemotaxis in
Escherichia coli
Authors: Adler Julius
Published: January 2020

Adler, Julius
(University of Wisconsin, Madison). Effect of amino acids and oxygen on chemotaxis in
Escherichia coli
. J. Bacteriol.
92:
121–129. 1966.—Motile cells of
Escherichia coli
placed at one end of a capillary tube containing a mixture of the 20 amino acids commonly found in proteins migrate out into the tube in two bands. The bands are clearly visible to the naked eye, and they can also be demonstrated by microscopy, photography, and densitometry, and by assaying for bacteria throughout the tube. The occurrence of more than one band is not due to heterogeneity among the bacteria, since each band can be used over to give rise to two again. The first band uses all the oxygen to oxidize portions of one or more of the amino acids, including serine, and the second band consumes the residual serine anaerobically. The results are interpreted to mean that
E. coli
shows chemotaxis toward oxygen and serine. When no energy source is added to the medium, a band of bacteria still appears. It consumes all the oxygen to oxidize an endogenous energy source. The addition of any one of 10 oxidizable amino acids stimulates the rate of travel of this band. Alanine, an example that was studied in detail, supports such a band that consumes all the oxygen to oxidize a portion of the alanine. Serine, the only amino acid that this strain can use either aerobically or anaerobically when grown under the conditions used here, gives rise to two bands.
Stability of the Lactobacillus Population in Feces and Stomach Contents of Rats Prevented from Coprophagy
Authors: Lev Meir , Alexander Raymond H. , Levenson Stanley M.
Published: January 2020

Lev, Meir
(Albert Einstein College of Medicine, New York, N.Y.),
Raymond H. Alexander, and Stanley M. Levenson
. Stability of the
Lactobacillus
population in feces and stomach contents of rats prevented from coprophagy. J. Bacteriol.
92:
13–16. 1966.—Lactobacilli were enumerated in the feces of rats prevented from coprophagy by tail-cupping. No differences were found when numbers of these organisms were compared with lactobacilli in feces of control rats, without tail-cups. High and similar numbers of lactobacilli were found in the stomachs of rats with and without tail-cups. The effect of coprophagy on fecal lactobacilli was therefore negligible.
Altered Sporulation and Respiratory Patterns in Mutants of
Bacillus subtilis
Induced by Acridine Orange
Authors: Bott K. F. , Davidoff-Abelson R.
Published: January 2020

Bott
, K. F. (The University of Chicago, Chicago, Ill.),
and R. Davidoff-Abelson
. Altered sporulation and respiratory patterns in mutants of
Bacillus subtilis
induced by acridine orange. J. Bacteriol.
92:
229–240. 1966.—The addition of acridine orange to vegetative cultures of
Bacillus subtilis
induces the formation of sporulation mutants at a frequency of 20% or greater. These mutants are grouped into seven categories which reflect their different morphological properties. They are altered in their vegetative metabolism, as indicated by abnormal growth on synthetic media. Sporulation of these mutants is impaired at several levels, all of which are stable upon repeated subculturing. The initial stages of sporulation which require no increased metabolic activity (proteolytic enzyme activity and antibiotic production) are functional in all strains, but glucose dehydrogenase activity, an enzyme associated with early synthetic functions in spore synthesis, is significantly reduced. Reduced nicotinamide adenine dinucleotide oxidase is slightly depressed. It is suggested that acridine orange interacts with a cellular constituent controlling respiration and consequently prevents an increased metabolic activity that may be associated with normal spore synthesis.
Fluorescence and Viability of
Proteus mirabilis
Stained Directly with Fluorescein Isothiocyanate
Authors: Grunberg E. , Cleeland R.
Published: January 2020

Grunberg
, E. (Hoffman-La Roche, Inc., Nutley, N.J.),
and R. Cleeland
. Fluorescence and viability of
Proteus mirabilis
stained directly with fluorescein isothiocyanate. J. Bacteriol.
92:
23–27. 1966.—Washed cell suspensions of
Proteus mirabilis
, under the proper conditions, stained well with fluorescein isothiocyanate with little or no loss of cell viability. The speed and intensity of the reaction was dependent on both the concentration of dye and
p
H. Within a range of
p
H 3.0 to 10.0, staining was most rapid at
p
H 5.0 to 6.0, with a slower and less intense reaction occurring at the other
p
H values. As the concentration of dye at either
p
H 5.0 or 9.0 was increased from 10 to 1,000 μg/ml, there was an increase in the rate of staining but a decrease in cell viability. After 24 hr of incubation at 4 C,
p
H 5.0, and a dye concentration of 10 μg/ml, all cells were stained, the majority exhibiting intense fluorescence with little or no loss of viability noted. In preliminary experiments with
Staphylococcus aureus
, similar results were obtained. Of various other fluorescent dyes tested, only rhodamine isothiocyanate was found to give satisfactory staining.
Suppression of Fluorophenylalanine Resistance by Mutation to Streptomycin Resistance in
Pseudomonas aeruginosa
Authors: Waltho Judith A. , Holloway B. W.
Published: January 2020

Waltho, Judith
A. (University of Melbourne, Victoria, Australia),
and B. W. Holloway
. Suppression of fluorophenylalanine resistance by mutation to streptomycin resistance in
Pseudomonas aeruginosa
. J. Bacteriol.
92:
35–42. 1966.—Fluorophenylalanine-resistant mutants (
fpa-r
) of
Pseudomonas aeruginosa
have been isolated. By cotransduction analysis, the mutations were shown to have at least two chromosomal locations. One locus (
fpaA
) showed linkage to three other markers,
str, try-3bi
, and
arg-3
, and the order of these four linked markers was found to be
try-3bi, arg-3, fpaA, str
. The linkage relationships of the other
fpa
loci are not yet known. The phenotypic expression of resistance at the
fpaA
locus can be suppressed by mutation of the
str
locus from
str-s
to
str-r
, whereas that at an unlinked
fpa
locus cannot.
Inhibition of Protein Synthesis by Spermine in Growing Cells of
Staphylococcus aureus
Authors: Friedman Mischa E. , Bachrach Uriel
Published: January 2020

Friedman, Mischa
E. (Hebrew University, Jerusalem, Israel),
and Uriel Bachrach
. Inhibition of protein synthesis by spermine in growing cells of
Staphylococcus aureus
. J. Bacteriol.
92:
49–55. 1966.—
Staphylococcus aureus
SFL 9725 incorporated valine-
C
14
into cellular protein. This incorporation was inhibited by spermine when the
p
H of the culture was adjusted to 7.8, and the inhibition was antagonized partially by Mg
++
. The incorporation of C
14
-labeled leucine, phenylalanine, lysine, arginine, and possibly glutamic acid was inhibited to a much greater extent than that of alanine, glycine, or threonine. The uptake of spermine-
C
14
by
S. aureus
cells was rapid. More than 50% of the radioactivity resided in the soluble extract. The protein fraction of the soluble extract contained 99% of the recovered label, whereas the ribonucleic acid and deoxyribonucleic acid fractions contained little or no spermine-
C
14
.
Polysaccharide Depolymerase Associated with Bacteriophage Infection
Authors: Bartell Pasquale F. , Orr Thomas E. , Lam Grace K. H.
Published: January 2020

Bartell, Pasquale
F. (University of Pennsylvania, Philadelphia),
Thomas E. Orr, and Grace K. H. Lam
. Polysaccharide depolymerase associated with bacteriophage infection. J. Bacteriol.
92:
56–62. 1966.—A recently isolated bacteriophage of
Pseudomonas aeruginosa
was observed, in association with bacteria, to produce a polysaccharide depolymerase. Exposure of slime polysaccharide to the enzyme at the
p
H optimum of 7.5 for 30 to 60 min resulted in a decreased viscosity of 20 to 25%, and a measurable increase in the levels of hexosamines, hexoses, and reducing substances, distinguishing it from other phage-associated depolymerases. Like egg-white lysozyme, the depolymerase produced a clearing of mature bacterial lawns, but was shown to be devoid of muralytic activity by turbidimetric and paper chromatographic analysis. The depolymerase reacted with polysaccharides of only certain strains of
P. aeruginosa
, and there appeared to be no correlation with phage susceptibility. The enzyme was not detectable in uninfected cultures, nor was it synthesized when infection was initiated by phages other than phage 2. The available data suggest that the genetic information required for biosynthesis of this enzyme is furnished by the phage 2 genome.
Nutrition and Metabolism of Marine Bacteria XV. Relation of Na
+
-Activated Transport to the Na
+
Requirement of a Marine Pseudomonad for Growth
Authors: Drapeau Gabriel R. , Matula Tibor I. , MacLeod Robert A.
Published: January 2020

Drapeau, Gabriel
R., (McGill University, Montreal, Quebec, Canada),
Tibor I. Matula, and Robert A. MacLeod
. Nutrition and metabolism of marine bacteria. XV. Relation of Na
+
-activated transport to the Na
+
requirement of a marine pseudomonad for growth. J. Bacteriol.
92:
63–71. 1966.—A marine pseudomonad was found to require 50 to 100 m
m
Na
+
for maximal rate of oxidation of
d
-galactose and for the transport of
d
-fucose-
H
3
into the cells. The same organism required 150 to 200 m
m
Na
+
for the oxidation of
l
-alanine and for the transport of φ-aminoisobutyric acid-
C
14
(AIB-
C
14
) into the cells. Competition studies indicated that
d
-galactose and
d
-fucose on the one hand and
l
-alanine and AIB on the other shared common carriers for transporting the compounds into the cells. This parallelism in Na
+
response for oxidation and transport extended to growth when
l
-alanine was the sole carbon source in the medium. When
d
-galactose was the sole carbon source, an amount of Na
+
equal to that with
l
-alanine was needed. KCN and dinitrophenol but not ouabain inhibited the uptake of AIB-
C
14
by the cells. K
+
in addition to Na
+
was required for transport, and both Mg
++
and either Cl
−
or Br
−
were stimulatory.
Photobacterium fischeri
was also found to require Na
+
specifically for the uptake of AIB-
C
14
by the cells.
Heterogeneity of the Conserved Ribosomal Ribonucleic Acid Sequences of
Bacillus subtilis
Authors: Doi Roy H. , Igarashi Richard T.
Published: January 2020

Doi, Roy
H. (University of California, Davis),
and Richard T. Igarashi
. Heterogeneity of the conserved ribosomal ribonucleic acid sequences of
Bacillus subtilis
. J. Bacteriol.
92:
88–96. 1966.—Hybrid formation was demonstrated between
Bacillus subtilis
ribosomal ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) from various bacterial species. The high degree of complementarity between
B. subtilis
ribosomal RNA and the DNA from
B. cereus
and
B. stearothermophilus
suggested a method to test whether the same RNA sequences were hybridizing with the DNA from these two species. Saturation studies with 16
S
and 23
S
RNA preparations from
B. subtilis
showed that a definite number of complementary sites was present in each DNA. Base composition analyses of the RNA in the hybrid demonstrated that ribosomal RNA sequences were involved. Hybrid competition studies revealed that
B. stearothermophilus
ribosomal RNA could compete totally against
B. subtilis
ribosomal RNA for
B. stearothermophilus
DNA, although it could compete only partially against the
B. subtilis
ribosomal RNA hybridizing with
B. cereus DNA
. These observations were made independently with both 16
S
and 23
S
ribosomal RNA preparations. These results revealed that different nucleotide sequences of
B. subtilis
ribosomal RNA were hybridizing with the DNA from
B. cereus
and
B. stearothermophilus
. Two possible interpretations of these results are: (i) different nucleotide sequences from a homogeneous ribosomal RNA population are hybridizing with heterologous DNA preparations, and (ii) ribosomal RNA cistrons are heterogeneous.
Genetic Differentiation by Nucleic Acid Homology I. Relationships AmongMycoplasmaSpecies of Man
Authors: Reich Paul R. , Somerson Norman L. , Hybner Carol J. , Chanock Robert M. , Weissman Sherman M.
Published: January 2020

Reich, PaulR. (National Institutes of Health, Bethesda, Md.),Norman L. Somerson, Carol J. Hybner, Robert M. Chanock, and Sherman M. Weissman. Genetic differentiation by nucleic acid homology. I. Relationships amongMycoplasmaspecies of man. J. Bacteriol.92:302-310. 1966.—Genetic relatedness among human mycoplasmas was evaluated by measuring the amount of nucleic acid hybrid retained on a membrane filter. Hybrids were formed from deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) derived from representative strains of seven serologically distinct humanMycoplasmaspecies. The results indicate that serologically distinct humanMycoplasmaspecies can also be distinguished by the homology techniques. Low-level cross-reactivity was observed among nucleic acids derived from the seven species. Genetic heterogeneity was demonstrated among three strains ofM. salivariumand between two strains ofM. oraletype 2. In contrast, comparison of three strains and three passage levels ofM. pneumoniaerevealed them to be indistinguishable.M. pneumoniaeappears to be the most distinct of all human mycoplasmas, as shown by both homology and the high buoyant density value of its DNA. Nucleic acids from mycoplasmas which had identical buoyant densities were in some cases differentiable. Mycoplasmas with different DNA buoyant densities were invariably distinguishable by the homology technique.
Synthesis of Pyridoxine by a Pyridoxal Auxotroph of
Escherichia coli
Authors: Dempsey Walter B.
Published: January 2020

Dempsey, Walter
B. (University of Florida, Gainesville). Synthesis of pyridoxine by a pyridoxal auxotroph of
Escherichia coli
. J. Bacteriol.
92:
333–337. 1966.—A pyridoxal auxotroph of
Escherichia coli
B produced pyridoxol and pyridoxol 5′-phosphate during starvation for pyridoxal. The identification of these compounds was made both by bioassay and by ion-exchange chromatography. Pyridoxol 5′-phosphate oxidase activity was absent in extracts of the auxotroph. The rate of synthesis of total pyridoxine by a pyridoxal-starved culture of this auxotroph was 6.0 × 10
−6
moles per mg per hr. Cellular content of pyridoxine was constant at 4.0 × 10
−10
moles/mg.
Leukocytic Response in Monkeys Challenged with Staphylococcal Enterotoxin
Authors: Sugiyama H. , McKissic E. M.
Published: January 2020

Sugiyama
, H. (University of Chicago, Chicago, Ill.),
and E. M. McKissic, Jr
. Leukocytic response in monkeys challenged with staphylococcal enterotoxin. J. Bacteriol.
92:
349–352. 1966—The feeding of staphylococcal enterotoxin to monkeys elicited a leukocytosis which was evident within 0.5 hr of challenge. The peak neutrophilic leukocytosis was reached in 3 hr, and then subsided so that leukocyte counts were normal within 28 hr. Each of the three serological types of enterotoxin tested induced the same effects. Intravenous injection of enterotoxin slightly above the emetic
ed
50
level produced an initial leukopenia followed by a neutrophilic leukocytosis which was maximal 9 or more hr postinjection. With smaller intravenous challenges, some animals responded with a leukopenia followed by a leukocytosis, some with only a leukocytosis, and others with no significant change in total leukocyte counts. The reversal of normal lymphocyte-to-neutrophile ratio toward a neutrophile-predominant white blood cell population occurred in all animals.
Effects of Dimethylsulfoxide on the Lactose Operon in
Escherichia coli
Authors: Fowler Audree V. , Zabin Irving
Published: January 2020

Fowler, Audree
V. (University of California, Los Angeles),
and Irving Zabin
. Effects of dimethylsulfoxide on the lactose operon in
Escherichia coli
. J. Bacteriol.
92:
353–357. 1966.—Dimethylsulfoxide (DMSO) at a concentration of 5% (v/v) in the culture medium inhibits the growth of
Escherichia coli
to only a slight extent, and does not affect the differential rate of synthesis of β-galactosidase. Resting cells remain viable after shaking in the presence of 20% DMSO for 3 hr at 37 C. Both β-galactosidase and thiogalactoside transacetylase retain almost all activity after incubation in even higher concentrations of the solvent for many hours. DMSO decreases the permeability barrier. The rate of hydrolysis of
o
-nitrophenyl-β-
d
-galactoside (ONPG) in whole cells containing β-galactosidase but lacking permease is increased in cells treated with 5% DMSO. Several permeaseless strains preinduced for β-galactosidase will grow on lactose in the presence, but not in the absence, of 5% DMSO. When permeaseless strains are grown on tetrazolium-lactose-agar, the presence of 5% DMSO causes a definite but not marked shift toward the lactose-positive character.
Mechanisms of Antibiotic Resistance Determined by Resistance-Transfer Factors
Authors: Unowsky Joel , Rachmeler Martin
Published: January 2020

Unowsky, Joel
(Northwestern University Medical School, Chicago, Ill.),
and Martin Rachmeler
. Mechanisms of antibiotic resistance determined by resistance-transfer factors. J. Bacteriol.
92:
358–365. 1966.—This study was concerned with the mechanism of expression of drug resistance carried by resistance-transfer (R) factors of two types:
fi
−
(negative fertility inhibition) and
fi
+
(positive fertility inhibition). The levels of drug resistance determined by R factors used in this study were similar to those reported by other investigators. A new finding was that
Escherichia coli
carrying the
fi
−
episome was resistant to 150 to 200 μg/ml of streptomycin. The growth kinetics of R factor-containing cells were similar in the presence or absence of streptomycin, chloramphenicol, and tetracycline, but a period of adaptation was necessary before cells began exponential growth in the presence of tetracycline. By use of radioactive antibiotics, it was shown that cells containing the
fi
−
episome were impermeable to tetracycline and streptomycin, whereas cells containing the
fi
+
episome were impermeable only to chloramphenicol. Cell-free extracts from
fi
+
and
fi
−
cells were sensitive to the antibiotics tested in the polyuridylic acid-stimulated incorporation of phenylalanine into protein.
Effect of Bromouracil-containing Deoxyribonucleic Acid on
Bacillus subtilis
Authors: Gimlin Dixie M. , Hardman Sue D. , Kelley Betty N. , Butler Grace C. , Leach Franklin R.
Published: January 2020

Gimlin, Dixie
M. (Oklahoma State University, Stillwater),
Sue D. Hardman, Betty N. Kelley, Grace C. Butler, and Franklin R. Leach
. Effect of bromouracil-containing deoxyribonucleic acid on
Bacillus subtilis
. J. Bacteriol.
92:
366–374. 1966.—Replacement of one-half of the thymine with bromouracil in
Bacillus subtilis
transforming deoxyribonucleic acid (DNA) resulted in a slight decrease in transforming activity, but, when used at high concentrations, this DNA preparation inhibited cell growth. Acid-hydrolyzed DNA, or addition of equivalent concentrations of the free base bromouracil in a transforming mixture, was without effect on cell growth. Treatment of the DNA preparation with deoxyribonuclease completely destroyed transforming activity and killing effect, whereas treatments with ribonuclease and trypsin were without effect on either transformation or killing activity. Growth of competent
B. subtilis
cells in test tubes was inhibited by high concentrations of both normal and bromouracil-containing DNA, with the bromouracil-containing DNA being significantly more inhibitory. This type of inhibition was also reflected in the time of division of the cells. The inhibitory effect was not due to viscosity, or to mutagenicity. The time course of killing paralleled transformation, and competency was required. These results can be interpreted as being due to uptake of homologous but imperfect DNA (containing bromouracil instead of thymine) by means of the systems involved in transformation, followed by either integration (resulting in lethal transformation, activation of a defective, nonlytic but lethal prophage) or interference with the recombination mechanism.
Role of Multivalent Cations in the Organization, Structure, and Assembly of the Cell Wall of
Pseudomonas aeruginosa
Authors: Asbell Mary A. , Eagon R. G.
Published: January 2020

Asbell, Mary
A. (University of Georgia, Athens),
and
R. G.
Eagon
. Role of multivalent cations in the organization, structure, and assembly of the cell wall of
Pseudomonas aeruginosa
. J. Bacteriol.
92:
380–387. 1966. —Incubation of
Pseudomonas aeruginosa
with ethylenediaminetetraacetate induced the formation of osmotically fragile rods termed osmoplasts. These could be restored to osmotically stable forms by multivalent cations. Only those cells restored by divalent cations normally found in the cell wall were capable of multiplication. The respiration of restored cells, however, was unimpaired, irrespective of whether they were capable of multiplication. Moreover, the permeability characteristics of osmoplasts and restored cells were unimpaired. When multivalent cations were chelated from the cell wall and replaced by sodium, a weakened cell wall and an osmotically fragile cell resulted. This was apparently caused by the absence of cross-linkages in the cell wall via multivalent cations. Tris(hydroxymethyl)aminomethane buffer compounded the lethal effects of ethylenediaminetetraacetate. The lipopolysaccharide component was inferred to be the site of attack by ethylenediaminetetraacetate. A mechanism for the synthesis of the lipopolysaccharide sacculus was proposed whereby negatively charged subunits are “trapped” by forming ionic and coordinate bonds intermediated by multivalent cations.
Properties of Nicotinamide Adenine Dinucleotide Phosphate-Dependent Formate Dehydrogenase from
Clostridium thermoaceticum
Authors: Li Lan-Fun , Ljungdahl Lars , Wood Harland G.
Published: January 2020

Li, Lan-Fun
(Western Reserve University School of Medicine, Cleveland, Ohio),
Lars Ljungdahl, and Harland G. Wood
. Properties of nicotinamide adenine dinucleotide phosphate-dependent formate dehydrogenase from
Clostridium thermoaceticum
. J. Bacteriol.
92:
405–412. 1966.—A nicotinamide adenine dinucleotide phosphate (NADP)-dependent formate dehydrogenase has been isolated from
C. thermoaceticum
. The enzyme is very sensitive to oxygen and requires sulfhydryl compounds for activity. The apparent
K
m
at 50 C and
p
H 7.0 for NADP is 5.9 × 10
−5
m
and for formate, 2.2 × 10
−4
m
. The enzyme is most active at about 60 C and at
p
H values between 7.0 and 9.0. The enzyme catalyzes an exchange between C
14
O
2
and formate, which requires NADP, but net synthesis of formate from CO
2
and reduced nicotinamide adenine dinucleotide phosphate could not be demonstrated. The reaction does not involve ferredoxin.
Thermolability of Malic Dehydrogenase from the Obligate Psychrophile
Vibrio marinus
Authors: Langridge Patricia , Morita Richard Y.
Published: January 2020

Langridge, Patricia
(Oregon State University, Corvallis),
and Richard Y. Morita
. Thermolability of malic dehydrogenase from the obligate psychrophile
Vibrio marinus
. J. Bacteriol.
92:
418–423. 1966.—The thermolability of malic dehydrogenase in whole cells of
Vibrio marinus
MP-1 grown at 15 C was compared with that of cell-free extracts and partially purified fractions. The intracellular enzyme was found to be stable between 0 C, and the organism's optimal growth temperature, 15 C. In cell-free extracts, considerable lability was noted even at 0 C, and this lability did not increase further until the enzyme was exposed to temperatures above the organism's maximal growth temperature (20 C). Twenty-fold purified enzyme was stable between 15 and 20 C, but both above and below this there was considerable inactivation. A 5-min exposure of both cold- and heat-inactivated enzyme to 15 C allowed reactivation, although to a different extent. Ammonium sulfate was found both to stimulate enzyme activity and to reactivate temperature-inactivated enzyme.
Genetic Recombination of Transforming Deoxyribonucleic Acid Molecules with the Recipient Genome and Among Themselves in Protoplasts of
Bacillus subtilis
Authors: Hirokawa Hideo , Ikeda Yonosuke
Published: January 2020

Hirokawa, Hideo
(Southwest Center for Advanced Studies, Dallas, Tex.),
and Yonosuke Ikeda
. Genetic recombination of transforming deoxyribonucleic acid molecules with the recipient genome and among themselves in protoplasts of
Bacillus subtilis
. J. Bacteriol.
92:
455–463. 1966.—Re-extraction of transforming deoxyribonucleic acid (DNA) from protoplasts of
Bacillus subtilis
is much more efficient than from intact competent cells. This facilitated the detection of physical recombination between donor and recipient DNA molecules, as indicated by a high cotransfer index of
ind
+
and
his
+
markers which were originally located in exogenous and endogenous DNA molecules, respectively. This recombinant DNA was extracted after 30 min of incubation of
ind his
+
protoplasts with
ind
+
his
DNA, previously extracted from a corresponding mutant strain of
B. subtilis
. The intracellular formation of recombinant molecules (
ind
+
his
+
) bearing markers from two different exogenous DNA species was also detected 15 min after exposure of
ind his
recipient protoplasts to a mixture of
ind
+
his
and
ind his
+
donor DNA molecules. The unity of the recombinant molecule was ascertained by dilution experiments and by its being resistant to ribonuclease and trypsin treatment (but being sensitive to deoxyribonuclease). The formation of recombinant molecules showed an inverse kinetics to that of the intracellularly induced loss of linkage between the corresponding markers in the wild-type DNA, thus suggesting a breakage and reunion process which is also favored by the absence of DNA synthesis in the protoplasts and the effect of some specific inhibitors.
Isolation of a Mutant of
Escherichia coli
with a Temperature-sensitive Fructose-1,6-Diphosphate Aldolase Activity
Authors: Böck August , Neidhardt Frederick C.
Published: January 2020

Böck, August
(Purdue University, Lafayette, Ind.),
and Frederick C. Neidhardt
. Isolation of a mutant of
Escherichia coli
with a temperature-sensitive fructose-1,6-diphosphate aldolase activity. J. Bacteriol.
92:
464–469. 1966.—A mutant of
Escherichia coli
was isolated which was able to grow in rich medium at 30 C but not at 40 C. Upon exposure to 40 C, the cells immediately stopped ribonucleic acid (RNA) and deoxyribonucleic acid synthesis, but protein synthesis continued at a diminished rate for a short time. Addition of chloramphenicol did not release RNA synthesis from inhibition at 40 C. Synthesis of β-galactosidase could be induced at high temperature despite the presence of glucose in the medium, indicating a lesion in glucose catabolism. Of many catabolic enzymes tested in cell-free extracts, only fructose-1,6-diphosphate aldolase activity appeared to be altered in the mutant cells. No activity was demonstrable in extracts of mutant cells grown at either 30 or 40 C, but determination of glucose-oxidation patterns revealed that the enzyme is probably active in vivo at 30 C. Temperature-resistant secondary mutants were found to have partially or fully restored aldolase activity, and temperature-resistant recombinants had normal aldolase activity, indicating that the growth pattern and the altered aldolase had a common genetic basis. Linkage data permitted the assignment of an approximate map location for the mutated aldolase gene.
Properties of a Mutant ofEscherichia coliwith a Temperature-sensitive Fructose-1,6-Diphosphate Aldolase
Authors: Böck August , Neidhardt Frederick C.
Published: January 2020

Böck, August(Purdue University, Lafayette, Ind.),and Frederick C. Neidhardt. Properties of a mutant ofEscherichia coliwith a temperature-sensitive fructose-1,6-diphosphate aldolase. J. Bacteriol.92:470–476. 1966.—A mutant ofEscherichia coliin which fructose-1,6-diphosphate aldolase functions at 30 C but not at 40 C was used to study the physiological effect of a specific block in the Embden-Meyerhof glycolytic pathway. Growth of the mutant at 40 C was found to be inhibited by the presence of glucose or certain related compounds in the medium. At 40 C, glucose was metabolized at 30 to 40% of the control rate and was abnormal in that glucose was converted into other six-carbon substances (probably gluconate, in large part) that were released into the culture medium. The inhibition was complete, but transient; its duration depended upon the initial amount of inhibitor added. The resumption of growth at 40 C was correlated with the further catabolism of the excreted compounds. When glycerol was used to grow the mutant at 40 C, the growth inhibition by glucose was accompanied by cessation of glycerol metabolism. Growth on α-glycerol phosphate was not inhibited under these conditions, implicating glycerol kinase as a possible site of inhibition; no inhibition of glycerol kinase by sugar phosphates, however, could be detected in vitro. The inhibitory effect of glucose on growth at 40 C is not caused by a deficit of intracellular adenosine triphosphate, but may be the result of a generalized poisoning of many cell processes by a greatly increased intracellular concentration of fructose-1,6-diphosphate, the substrate of the damaged enzyme.
Lethal Gram-Negative Bacterial Superinfection in Guinea Pigs Given Bacitracin
Authors: Farrar W. Edmund , Kent Thomas H. , Elliott Van B.
Published: January 2020

Farrar, W. Edmund, Jr.
(Walter Reed Army Institute of Research, Washington, D.C.),
Thomas H. Kent, and Van B. Elliott
. Lethal gram-negative bacterial superinfection in guinea pigs given bacitracin. J. Bacteriol.
92:
496–501. 1966.—Oral administration of a single dose of bacitracin (either 2,000 or 10,000 units) was lethal to more than 80% of guinea pigs. Within the first 12 hr, there was a 2,000-fold fall in the number of gram-positive organisms in the cecum. An increase in the number of coliform bacteria in the cecum was demonstrable within 6 hr, and, by 48 hr, these organisms had increased from the normal level of less than 100 per gram to approximately 1 billion per gram. The changes in intestinal bacterial flora were associated with development of a severe cecitis, mild ileitis, and acute regional lymphadenitis. Bacteremia, primarily due to coliform bacteria, was demonstrated in approximately 40% of the animals killed between 72 and 96 hr after administration of bacitracin. Development of this disease syndrome was suppressed by the administration of neomycin and polymyxin B, nonabsorbable antibiotics effective against coliform bacteria. The lethal disease produced by bacitracin in the guinea pig is similar to that produced by penicillin.
Mechanisms of Pathogenesis inListeria monocytogenesInfection I. Influence of Iron
Authors: Sword C. P.
Published: January 2020

Sword, C. P. (The University of Kansas, Lawrence). Mechanisms of pathogenesis inListeria monocytogenesinfection. I. Influence of iron. J. Bacteriol.92:536–542. 1966.—The effects of ferric and ferrous iron as well as other cations onListeriainfection in mice were studied. Iron compounds caused a reduction in theld50dose ofListeria, and, when added to a synthetic medium, proved stimulatory for in vitro growth of the organism. Bacterial counts on spleen and liver tissue from iron-treated mice showed that iron injections caused more rapid growth of bacteria and resulted in higher numbers of organisms in the tissue. The reticuloendothelial system did not appear to be impaired by this treatment. Immunized animals were not affected by iron treatment during challenge. Mice with experimentally induced hemolytic anemia showed increased susceptibility to listeriosis, whereas those treated with Desferal, a specific iron-chelating agent, appeared more resistant. Iron proved stimulatory for the avirulent strain, 9037–7, and resulted in anld50of 1.3 × 104organisms in iron-treated animals. Growth ofL. monocytogenesand mortality from experimental infection appeared to be correlated with availability of iron to the bacteria. The results suggest that host iron metabolism may play a part in the onset and progress ofListeriainfections.
Isotopic Method for Estimating Microbial Cell Volumes
Authors: Reid Della F. , Frank Hilmer A.
Published: January 2020

Reid, Della
F. (University of Hawaii, Honolulu),
and Hilmer A. Frank
. Isotopic method for estimating microbial cell volumes. J. Bacteriol.
92:
639–644. 1966.—A sensitive isotopic method was developed for estimating the mean cell volume in small numbers of bacterial spores by measuring directly the extracellular fluid volume when cells are present in a saturated inulin solution containing inulin-C
14
at low specific activity. The method minimizes errors due to solute adsorption to the cells and avoids errors arising from differences in cell packing. The effects of these sources of error on cell volume and intercellular space estimations made with nonpenetrating solutes are discussed.
In Vitro Interactions Between Rabbit Alveolar Macrophages andPasteurella tularensis
Authors: Nutter J. E. , Myrvik Q. N.
Published: January 2020

Nutter, J. E. (Fort Detrick, Frederick, Md.),and Q. N. Myrvik. In vitro interactions between rabbit alveolar macrophages andPasteurella tularensis. J. Bacteriol.92:645–651. 1966.—Rabbit alveolar macrophages were successfully employed in a study of host cell-Pasteurella tularensisinteractions in vitro. Under cell culture conditions in which inhibitory antibiotics were not employed and small infection ratios were used, the relative in vivo virulence of two strains ofP. tularensiswas duplicated. As a consequence of intracellular multiplication, normal macrophages were killed in relation to the virulence of the strain employed. Alveolar macrophages were also collected from immune rabbits, and macrophage mortality and bacterial growth were significantly suppressed below levels observed with normal macrophage preparations. The effect of immune serum could only be ascribed a minor role in the observed reactions. A marked intravenous toxicity ofP. tularensisfor the rabbit was observed with both virulent and attenuated strains. The toxicity was possessed only by viable preparations and could be elicited in animals immune to virulent challenge.
Occurrence of Hydroxypyruvate—
l
-Glutamate Transaminase in
Escherichia coli
and Its Separation from Hydroxypyruvate-Phosphate—
l
-Glutamate Transaminase
Authors: Blatt L. , Dorer F. E. , Sallach H. J.
Published: January 2020

Blatt
, L. (University of Wisconsin, Madison), F. E.
Dorer, and H. J. Sallach
. Occurrence of hydroxypyruvate–
l
-glutamate transaminase in
Escherichia coli
and its separation from hydroxypyruvate-phosphate–
l
-glutamate transaminase. J. Bacteriol.
92:
668–675. 1966.—The formation of
l
-serine from hydroxypyruvate by a transamination reaction with
l
-glutamate has been demonstrated in extracts of
Escherichia coli
. The level of activity with hydroxypyruvate is approximately one-tenth that observed with hydroxypyruvate-phosphate in cell-free extracts. The transamination of hydroxypyruvate, but not hydroxypyruvate-phosphate, is inhibited by inorganic phosphate. No marked differences in the levels of activity with hydroxypyruvate were observed in extracts from bacteria grown under different conditions. Heat treatment of enzyme preparations at 65 C rapidly destroys the activity with hydroxypyruvate-phosphate, but not that with hydroxypyruvate. Fractionation of extracts with lithium sulfate and alumina Cγ resulted not only in a 10-fold purification, but also in a complete separation of the two activities, thereby establishing that two different enzymes are involved in the transamination of hydroxypyruvate and hydroxypyruvate-phosphate. Hydroxypyruvate transaminase is present in two mutants that require serine for growth. The inability of hydroxypyruvate to replace the growth requirement for serine, even to a limited extent, was shown to be due to the inability of the bacteria to accumulate this compound actively.
Endogenous Metabolism of
Azotobacter agilis
Authors: Sobek J. M. , Charba J. F. , Foust W. N.
Published: January 2020

Sobek
, J. M. (University of Southwestern Louisiana, Lafayette), J. F.
Charba, and
W. N.
Foust
. Endogenous metabolism of
Azotobacter agilis
. J. Bacteriol.
92:
687–695. 1966—Ribonucleic acid, deoxyribonucleic acid, cellular carbohydrate, and the cold trichloroacetic acid and acidic alcohol fractions of the cell do not appear to function as endogenous reserves for
Azotobacter agilis
. The immediate endogenous reserve of cells grown on glucose, acetate, or succinate was poly-β-hydroxybutyric acid (PHB). Viability of the cells during starvation was dependent upon the initial levels of PHB and the growth substrate. Cells with high initial PHB levels survived longer than cells with lower levels. Cells from succinate-grown cultures had lower PHB levels than cells from glucose-grown cultures, but were capable of maintaining their viability longer. Cellular protein may also serve as a secondary endogenous reserve substrate for this organism.
Conversion of Mevalonic Acid to γ,γ-Dimethylallyl Pyrophosphate by
Mycoplasma
Authors: Henrikson Carl V. , Smith Paul F.
Published: January 2020

Henrikson, Carl
V. (University of South Dakota, Vermillion),
and Paul
F.
Smith
. Conversion of mevalonic acid to γ,γ-dimethylallyl pyrophosphate by
Mycoplasma
. J. Bacteriol.
92:
701–706. 1966.—Three representative strains of
Mycoplasma, M. laidlawii
strain B,
Mycoplasma
sp. avian strain J, and
M. hominis
type 2 strain O7, were examined for the presence or absence of enzymes associated with the biosynthetic pathway from mevalonic acid to γ,γ-dimethylallyl pyrophosphate.
M. laidlawii
served as a control organism, since it is capable of de novo biosynthesis of carotenoids. All four enzymes, namely, adenosine triphosphate (ATP)-mevalonate 5-phosphotransferase (EC 2.7.1.36), ATP-5-phosphomevalonate phosphotransferase (EC 2.7.4.2), ATP-5-pyrophosphomevalonate carboxy-lyase (EC 4.1.1.33), and isopentenylpyrophosphate Δ
3
,Δ
2
-isomerase (EC 5.3.3.2), were demonstrated in this organism.
Mycoplasma
sp. avian strain J, which contains all enzymes necessary for the biosynthesis of mevalonic acid, lacks the first three of the above enzymes but contains isopentenyl pyrophosphate Δ
3
,Δ
2
-isomerase.
M. hominis
, which lacks the enzymes necessary for the biosynthesis of mevalonic acid, also is deficient in the enzymes involved in its conversion to γ,γ-dimethylallyl pyrophosphate.
Adenosine Triphosphatase Activity of Mycoplasma Membranes
Authors: Rottem Shlomo , Razin Shmuel
Published: January 2020

Rottem, Shlomo
(Hebrew University, Jerusalem, Israel),
and Shmuel Razin
. Adenosine triphosphatase activity of mycoplasma membranes. J. Bacteriol.
92:
714–722. 1966.—Adenosine triphosphatase activity of
Mycoplasma laidlawii, M. gallisepticum
, and
Mycoplasma
sp. strain 14 was confined to the cell membrane. The enzymatic activity was dependent on magnesium, but was not activated by sodium and potassium. Ouabain did not inhibit the adenosine triphosphatase activity of the mycoplasmas, and did not interfere with the active accumulation of potassium by
M. laidlawii
cells. Sulfhydryl-blocking reagents and fluoride inhibited the enzymatic activity, whereas 2,4-dinitrophenol was without any effect. Membranes of
M. laidlawii
hydrolyzed other nucleotide triphosphates and adenosine diphosphate (ADP), but at a lower rate than adenosine triphosphate (ATP). Nucleoside-2′-(3′)-phosphates, ribose-5-phosphate, glucose-6-phosphate, and pyrophosphate were not hydrolyzed by the membrane preparations. It seems that the enzyme(s) involved in ATP hydrolysis by
M. laidlawii
membranes is strongly bound to the membrane subunits, which would account for the failure to purify the enzyme by protein fractionation techniques. The adenosine triphosphatase activity of mycoplasma membranes resembles in its properties that of similar enzymes studied in bacteria. The mycoplasma enzyme(s) seems to differ from the adenosine triphosphatase associated with ion transport in mammalian cell membranes and from mitochondrial adenosine triphosphatase.
Quantitative Aspects of Inhibition of Virus Replication by Interferon in Chick Embryo Cell Cultures
Authors: Hallum J. V. , Youngner J. S.
Published: January 2020

Hallum, J. V. (University of Pittsburgh, Pittsburgh, Pa.),and J. S. Youngner. Quantitative aspects of inhibition of virus replication by interferon in chick embryo cell cultures. J. Bacteriol.92:1047–1050. 1966.—The effect of interferon on single cycles of replication of vesicular stomatitis virus and Mahoney poliovirus ribonucleic acid was studied in chick embryo cell cultures. The results showed that the titer of a given interferon preparation was independent of the input multiplicity of the challenge virus. In addition, the increase in virus yield with increasing virus challenge was a function of the number of infected cells, each of which yielded progeny at a level determined by the concentration of interferon to which the cells were exposed. These findings are not compatible with the concept that increases in the size of the virus challenge reverse or overcome protection of cells by interferon.
Ultraviolet Radiation Studies of FilamentousEscherichia coliB
Authors: Kantor George J. , Deering R. A.
Published: January 2020

Kantor, GeorgeJ. (The Pennsylvania State University, University Park),and R. A. Deering. Ultraviolet radiation studies of filamentousEscherichia coliB. J. Bacteriol.92:1062–1069. 1966.—Small ultraviolet (UV) doses causeEscherichia coliB to grow into long filamentous single cells. A large fraction of these filaments can recover their division ability and can form colonies under appropriate conditions. Preformed filaments can be irradiated with UV, and their ability to still produce colonies can be compared with that of irradiated normal cells. In this regard, filaments are more sensitive to UV than normal cells. Filaments can still host-cell reactivate UV-irradiated T1 phage and can regain their own deoxyribonucleic acid (DNA) synthetic ability after it has been blocked by UV. This indicates that these filaments still retain mechanisms for repairing UV-damaged DNA. Pantoyl lactone, an agent that stimulates cell-division recovery in UV-irradiatedE. coliB, causes increased UV resistance for both normal and filamentous cells, with the filaments becoming more resistant than normal cells. In the absence of pantoyl lactone, irradiated filaments grow to a length of about 50 times normal and then stop growing. These long filaments cannot subsequently divide and give colonies. We conclude that the UV dose given to the preformed filaments causes an additional division lag beyond that of unirradiated filaments, and that some critical length is reached after which division recovery and colony formation is impossible. Irradiated normal cells recover before reaching this critical length.
Bacteriocins of
Diplococcus pneumoniae
I. Antagonistic Relationships and Genetic Transformations
Authors: Mindich Leonard
Published: January 2020

Mindich, Leonard
(The Public Health Research Institute of the City of New York, Inc., New York, N.Y.). Bacteriocins of
Diplococcus pneumoniae
. I. Antagonistic relationships and genetic transformations. J. Bacteriol.
92:
1090–1098. 1966.—Several strains of
Diplococcus pneumoniae
produced bacteriocins inhibiting the growth of other strains of pneumococcus but inactive upon themselves. Strain R6, which did not produce pneumocin under most conditions, was changed to a producing state by mutation or by transformation with deoxyribonucleic acid from other pneumococci. The genetic determinants of several different bacteriocins were introduced into the same cell, where they acted independently. Determinants active in bacteriocin production were also constructed by crosses involving two nonproducing parents. Resistance to pneumocin B4 was a property of cells of the P4 type which produced it. On selection of pneumocin-resistant cells resulting from mutation or transformation in strain R6, resistant cells, all of which were pneumocin producers, were obtained. The genetic determinants of B4 production and resistance were transferred together in transformation. A gene for high level B4 resistance was found in strain P7S. This gene was not associated with B4 production and did not induce production when introduced into R6, although it did increase resistance to B4. Pneumocin B4 was bactericidal and caused a rapid cessation of growth. Its action was inhibited by pronase, trypsin, chymotrypsin, and phospholipids.