bluuebunny/crossref_metadata_2025 huggingface dataset:
Kinetics of uptake and incorporation of meso-diaminopimelic acid in different Escherichia coli strains
Authors: Wientjes F B , Pas E , Taschner P E , Woldringh C L
Published: January 2020

The rate at which the peptidoglycan precursor meso-diaminopimelic acid (DAP) is incorporated into the cell wall of Escherichia coli cells was determined by pulse-label experiments. For different E. coli strains, the incorporation rate was compared with the rate of uptake of DAP into the cell. With E. coli W7, a dap lys mutant generally used in this kind of studies, steady-state incorporation was reached only after about 0.75 of the doubling time. This lag period can be ascribed to the presence of a large internal DAP pool in the cells. An E. coli K-12 lysA strain was constructed which could be grown without DAP in its medium. Consequently, due to the higher specific activity of the added [3H]DAP, faster incorporation and higher levels of radioactivity in the peptidoglycan layer were observed in the K-12 lysA strain than in the W7 strain. In addition, uptake and incorporation were faster in steady state (within about 0.2 of the doubling time), indicating a smaller DAP pool. The lag period could be further diminished and the incorporation rate could be increased by feedback inhibition of the biosynthetic pathway to DAP with threonine and methionine. These results make MC4100 lysA a suitable strain for studies on peptidoglycan synthesis. To explain our observations, we suggest the existence of an expandable pool of DAP in E. coli which varies with the DAP concentration in the growth medium. With 2 microgram of DAP per ml, the size of the pool is severalfold the amount of DAP contained in the cell wall. This pool can be partly washed out of the cells. Grown without DAP, MC4100 lysA still has a small pool caused by endogenous synthesis, which accounts for the fact that steady-state [3H]DAP incorporation in the lysA strain still shows a lag period.
Carotenoid-containing outer membrane of Synechocystis sp. strain PCC6714
Authors: Jürgens U J , Weckesser J
Published: January 2020

Outer membranes, free of cytoplasmic or thylakoid membranes and peptidoglycan components, were obtained from Synechocystis sp. strain PCC6714. Electron microscope studies revealed double-track outer membrane vesicles with a smooth-appearing exoplasmic surface, an exoplasmic fracture face covered by closely packed particles and a corresponding plasmic fracture face with regularly distributed holes. Lipopolysaccharide, proteins, lipids, and carotenoids were the constituents of the outer membrane of Synechocystis sp. PCC6714. Twelve polypeptides were found in outer membrane fractions, among them two dominant outer membrane proteins (Mrs, 67,000 and 61,000). Lipopolysaccharide-specific components were GlcN and an unidentified heptose. Outer membrane lipid extracts contained phosphatidylglycerol, sulfolipid, phosphatidylcholine, and unknown lipids. The carotenoids, myxoxanthophyll, related carotenoid-glycosides, zeaxanthin, echinenone, and beta-carotene were found to be true constituents of the outer membrane of Synechocystis sp. PCC6714.
Mutations affecting gyrase in Haemophilus influenzae
Authors: Setlow J K , Cabrera-Juárez E , Albritton W L , Spikes D , Mutschler A
Published: January 2020

Mutants separately resistant to novobiocin, coumermycin, nalidixic acid, and oxolinic acid contained gyrase activity as measured in vitro that was resistant to the antibiotics, indicating that the mutations represented structural alterations of the enzyme. One Novr mutant contained an altered B subunit of the enzyme, as judged by the ability of a plasmid, pNov1, containing the mutation to complement a temperature-sensitive gyrase B mutation in Escherichia coli and to cause novobiocin resistance in that strain. Three other Novr mutations did not confer antibiotic resistance to the gyrase but appeared to increase the amount of active enzyme in the cell. One of these, novB1, could only act in cis, whereas a new mutation, novC, could act in trans. An RNA polymerase mutation partially substituted for the novB1 mutation, suggesting that novB1 may be a mutation in a promoter region for the B subunit gene. Growth responses of strains containing various combinations of mutations on plasmids or on the chromosome indicated that low-level resistance to novobiocin or coumermycin may have resulted from multiple copies of wild-type genes coding for the gyrase B subunit, whereas high-level resistance required a structural change in the gyrase B gene and was also dependent on alteration in a regulatory region. When there was mismatch at the novB locus, with the novB1 mutation either on a plasmid or the chromosome, and the corresponding wild-type gene present in trans, chromosome to plasmid recombination during transformation was much higher than when the genes matched, probably because plasmid to chromosome recombination, eliminating the plasmid, was inhibited by the mismatch.
Purification, characterization, and immunological properties of fumarase from Euglena gracilis var. bacillaris
Authors: Shibata H , Gardiner W E , Schwartzbach S D
Published: January 2020

A rapid three-step procedure utilizing heat treatment, ammonium sulfate fractionation, and affinity chromatography on Matrex gel Orange A purified fumarase (EC 4.2.1.2) 632-fold with an 18% yield from crude extracts of Euglena gracilis var. bacillaris. The apparent molecular weight of the native enzyme was 120,000 as determined by gel filtration on Sephacryl S-300. The preparation was over 95% pure, and the subunit molecular weight was 60,000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, indicating that the enzyme is a dimer composed of two identical subunits. The pH optimum for E. gracilis fumarase was 8.4. The Km values for malate and fumarate were 1.4 and 0.031 mM, respectively. Preparative two-dimensional gel electrophoresis was used to further purify the enzyme for antibody production. On Ouchterlony double-immunodiffusion gels, the antifumarase serum gave a sharp precipitin line against total E. gracilis protein and purified E. gracilis fumarase. It did not cross-react with purified pig heart fumarase. On immunoblots of purified E. gracilis fumarase and crude cell extracts of E. gracilis, the antibody recognized a single polypeptide with a molecular weight of approximately 60,000, indicating that the antibody is monospecific. This polypeptide was found in E. gracilis mitochondria. The antibody cross-reacted with an Escherichia coli protein whose molecular weight was approximately 60,000, the reported molecular weight of the fumA gene product of E. coli, but it failed to cross-react with proteins found in crude mouse cell extracts, Bacillus subtilis extracts, or purified pig heart fumarase.
Regulation of Ti plasmid virulence genes by a chromosomal locus of Agrobacterium tumefaciens
Authors: Close T J , Tait R C , Kado C I
Published: January 2020

We isolated a mutant strain of Agrobacterium tumefaciens, designated Ros, that has a pleiotropic phenotype which includes elevated levels of expression of certain genes in the virulence (Vir) region of tumor-inducing plasmid pTiC58. This mutant and others were isolated by fusing the promoter of the Vir bak gene to a promoterless gene (cat) that encodes chloramphenicol acetyltransferase and then selecting for increased expression of cat in A. tumefaciens. The ros mutation is chromosomal in nature and is characterized by a more-than-300-fold increase in the level of expression of bak and a 12-fold increase in the level of expression of an adjacent divergent operon containing the hdv genes, which are involved in some aspect of host specificity. The Ros mutant is fully virulent and is typified by its unusual colony morphology; the colonies have rough surfaces, uneven edges, and a pit in the center at 24 degrees C and vary markedly in appearance from one growth temperature to another. The Ros mutant is not able to form colonies at 12 degrees C, a temperature at which the wild-type strain forms colonies in 14 days. The ros mutation occurs spontaneously with a frequency of 5 X 10(-8). Genetic and biochemical evidence indicates that the product of the ros locus is a negative regulator of Ti plasmid genes and is related to undefined chromosomally encoded functions that are involved in the mutant phenotype.
Differential amylosaccharide metabolism of Clostridium thermosulfurogenes and Clostridium thermohydrosulfuricum
Authors: Hyun H H , Shen G J , Zeikus J G
Published: January 2020

Clostridium thermosulfurogenes displayed faster growth on either glucose, maltose, or starch than Clostridium thermohydrosulfuricum. Both species grew faster on glucose than on starch or maltose. The fermentation end product ratios were altered based on higher ethanol and lactate yields on starch than on glucose. In C. thermohydrosulfuricum, glucoamylase, pullulanase, and maltase were mainly responsible for conversion of starch and maltose into glucose, which was accumulated by a putative glucose permease. In C. thermosulfurogenes, beta-amylase was primarily responsible for degradation of starch to maltose, which was accumulated by a putative maltose permease and then hydrolyzed by glucoamylase. Regardless of the growth substrate, the rates of glucose, maltose, and starch transformation were higher in C. thermosulfurogenes than in C. thermohydrosulfuricum. Both species had a functional Embden-Meyerhof glycolytic pathway and displayed the following catabolic activities: ferredoxin-linked pyruvate dehydrogenase, acetate kinase, NAD(P)-ethanol dehydrogenase, NAD(P)-ferredoxin oxidoreductase, hydrogenase, and fructose-1,6-diphosphate-activated lactate dehydrogenase. Ferredoxin-NAD reductase activity was higher in C. thermohydrosulfuricum than NADH-ferredoxin oxidase activity, but the former activity was not detectable in C. thermosulfurogenes. Both NAD- and NADP-linked ethanol dehydrogenases were unidirectional in C. thermosulfurogenes but reversible in C. thermohydrosulfuricum. The ratio of hydrogen-producing hydrogenase to hydrogen-consuming hydrogenase was higher in C. thermosulfurogenes. Two biochemical models are proposed to explain the differential saccharide metabolism on the basis of species enzyme differences in relation to specific growth substrates.
Cloning and expression of the Thiobacillus ferrooxidans glutamine synthetase gene in Escherichia coli
Authors: Barros M E , Rawlings D E , Woods D R
Published: January 2020

The glutamine synthetase (GS) gene glnA of Thiobacillus ferrooxidans was cloned on recombinant plasmid pMEB100 which enabled Escherichia coli glnA deletion mutants to utilize (NH4)2SO4 as the sole source of nitrogen. High levels of GS-specific activity were obtained in the E. coli glnA deletion mutants containing the T. ferrooxidans GS gene. The cloned T. ferrooxidans DNA fragment containing the glnA gene activated histidase activity in an E. coli glnA glnL glnG deletion mutant containing the Klebsiella aerogenes hut operon. Plasmid pMEB100 also enabled the E. coli glnA glnL glnG deletion mutant to utilize arginine or low levels of glutamine as the sole source of nitrogen. There was no detectable DNA homology between the T. ferrooxidans glnA gene and the E. coli glnA gene.
RELATIONSHIP BETWEEN THYMINELESS DEATH AND ULTRAVIOLET INACTIVATION IN
ESCHERICHIA COLI
Authors: Gallant J. , Suskind S. R.
Published: January 2020

Gallant
, J. (Johns Hopkins University, Baltimore, Md.),
and S. R. Suskind
. Relationship between thymineless death and ultraviolet inactivation in
Escherichia coli
. J. Bacteriol.
82:
187–194. 1961.—At 37 C, cultures of
Escherichia coli
strain B3 growing in the absence of thymine undergo unbalanced growth and thymineless death. Thymineless death also occurs at 37 C in 2 μg of chloramphenicol per ml despite virtually complete inhibition of cell growth and net protein synthesis. Thymineless cultures growing at 25 C exhibit unbalanced synthesis of ribonucleic acid and of protein without lethality. Examination of the kinetics of ultraviolet inactivation of cells grown under these three conditions indicates that the mean lethal dose of ultraviolet irradiation decreases with unbalanced protein synthesis whether or not thymineless death occurs, whereas the latter is specifically correlated with a transition from multihit to single hit inactivation kinetics.

The time course of this transition correlates well with that of thymineless death, and preirradiated cells show a shorter lag period before the onset of thymineless death. These results, therefore, suggest that thymineless death is due to an inactivation of those cellular units which are sensitive to ultraviolet irradiation, presumably nuclei or some nuclear function.
REPRESSION AND INHIBITION OF INDOLE-SYNTHESIZING ACTIVITY IN
NEUROSPORA CRASSA
Authors: Lester Gabriel
Published: January 2020

Lester, Gabriel
(Worcester Foundation for Experimental Biology, Shrewsbury, Mass.). Repression and inhibition of indole-synthesizing activity in
Neurospora crassa
. J. Bacteriol.
82:
215–223. 1961.—The possibility of repression and feedback inhibition as regulating mechanisms for the synthesis of tryptophan by
Neurospora crassa
has been examined in a tryptophan auxotroph which accumulates indole (and indole-glycerol). Indole-synthesizing activity was determined with germinated conidia suspended in medium lacking tryptophan. This activity was almost absent from cells cultured on germination medium containing more than 1.0 μmole
l
-tryptophan per ml, and increased with decreasing concentrations of
l
-tryptophan. A similar depression of the formation of indole synthesizing activity was caused by 6-methyl- and
d
-tryptophan, and less effectively by 5-methyltryptophan; 4-methyltryptophan was slightly stimulatory. Preformed indole synthesizing activity was inhibited by
l
-tryptophan, 4- and 6-methyltryptophan, and to a lesser extent by 5-methyltryptophan;
d
-tryptophan had no effect in this respect. The inhibition of preformed activity was partially reversed by anthranilic acid, which is a precursor of indole. However, anthranilic acid did not increase indole synthesis by cells wherein the formation of indole-synthesizing activity had been depressed by culture in the presence of high concentrations of
l
- or
d
-tryptophan. These observations indicate that regulation of tryptophan synthesis in
N. crassa
might result from the action of tryptophan as a repressor and as a feedback inhibitor. The relation of these results to other regulatory systems is discussed.
POLYOL DEHYDROGENASES OF
AZOTOBACTER AGILIS
Authors: Marcus Leon , Marr Allen G.
Published: January 2020

Marcus, Leon
(University of California, Davis),
and Allen G. Marr
. Polyol dehydrogenases of
Azotobacter agilis
. J. Bacteriol.
82:
224–232. 1961.—Two soluble diphosphopyridine-linked polyol dehydrogenases are formed by
Azotobacter agilis
(
A. vinelandii
). The first,
d
-mannitol dehydrogenase is induced by
d
-mannitol and all of the pentitols except
l
-arabitol. Ribitol is an excellent inducer of mannitol dehydrogenase although it is not metabolized, nor does the enzyme act upon it. This allows study of the gratuitous induction of mannitol dehydrogenase.

Of the polyols tested, mannitol dehydrogenase oxidizes
d
-mannitol,
d
-arabitol,
d
-rhamnitol, and perseitol, demonstrating its requirement for substrates bearing the
d
-
manno
configuration. The corresponding 2-ketoses,
d
-fructose,
d
-xylulose, and presumably
d
-rhamnulose, and perseulose are reduced.

The second enzyme,
l
-iditol dehydrogenase is induced only by polyols containing the
d
-
xylo
configuration, i.e., sorbitol and xylitol.
l
-Iditol dehydrogenase oxidizes
d
-
xylo
polyols seven times faster than it does
d
-
ribo
polyols. Substrates oxidized include
l
-iditol, sorbitol, xylitol, and ribitol. The corresponding 2-ketoses,
l
-sorbose,
d
-fructose,
d
-xylulose, and
d
-ribulose, are reduced.

The two polyol dehydrogenases have been separated and purified by chromatography on a modified cellulose ion exchanger.
EXPERIMENTAL SHIGELLA INFECTIONS V
Authors: Formal Samuel B. , Dammin Gustave , Sprinz Helmuth , Kundel Donald , Schneider Herman , Horowitz Richard E. , Forbes Martin
Published: January 2020

Formal, SamuelB., (Walter Reed Army Institute of Research, Washington, D. C.),Gustave Dammin, Helmuth Sprinz, Donald Kundel, Herman Schneider, Richard E. Horowitz, and Martin Forbes. Experimental shigella infections. V. Studies in germ-free guinea pigs. J. Bacteriol.82:284–287. 1961.—Germ-free guinea pigs succumb after oral infection withShigella flexneriserotype 2a; they survive a similar challenge with either a strain ofEscherichia colior a culture of lactobacillus. Animals monocontaminated withE. colisurvive, whereas those monocontaminated with lactobacilli succumb to subsequent challenge with dysentery bacilli. Prior subcutaneous inoculation of heat-killedS. flexneri2a does not render germ-free guinea pigs resistant to the fatal infection with viable dysentery bacilli.
COMPETITIVE RELATIONSHIP BETWEEN PROTOCATECHUIC ACID AND
p
-AMINOSALICYLIC ACID FOR A CELLULAR TRANSPORT MECHANISM
Authors: Hubbard Jerry S. , Durham Norman N.
Published: January 2020

Hubbard, Jerry
S. (Oklahoma State University, Stillwater),
and Norman N. Durham
. Competitive relationship between protocatechuic acid and
p
-aminosalicylic acid for a cellular transport mechanism. J. Bacteriol.
82:
361–369. 1961.—The oxidation of protocatechuic acid by a Flavobacterium is inhibited by
p
-aminosalicyclic acid regardless of whether the organism is grown on protocatechuic acid or sequentially induced to protocatechuic acid by growth on
p
-aminobenzoic acid. Depletion of the substrate from the medium by the cell suspension is dependent, within defined limits, on the inhibitor to substrate ratio, and the inhibition can be overcome by addition of excess substrate. However, this competitive effect is not observed in high inhibitor to substrate ratios.
p
-Aminosalicylic acid did not affect the rate or extent of oxidation, carbon dioxide evolution, or formation of β-ketoadipic acid during degradation of protocatechuic acid by cell extracts. The results suggest that
p
-aminosalicylic acid antagonizes the oxidation of protocatechuic acid by the cell suspension by competing with the substrate for a specific transport mechanism, thereby regulating the entry and internal accumulation of the substrate. The lack of a competitive effect in high inhibitor to substrate ratios could be interpreted as an indication that the mechanism for accumulating the substrate may consist of more than one active transport system.
ISOLATION AND CHARACTERIZATION OF DEOXYRIBONUCLEIC ACID FROM A STRAIN OF
STAPHYLOCOCCUS AUREUS
Authors: Blobel Hans
Published: January 2020

Blobel, Hans
(University of Wisconsin, Madison). Isolation and characterization of deoxyribonucleic acid from a strain of
Staphylococcus aureus
. J. Bacteriol.
82:
425–429. 1961.—Highly polymerized deoxyribonucleic acid was extracted from washed cells of
Staphylococcus aureus
with a mixture of equal parts of phenol and 2
m
NaCl at pH 7.4. The aqueous phase was treated twice with phenol and the deoxyribonucleic acid precipitated with an equal volume of 2-ethoxyethanol. Residual ribonucleic acid was removed by treatment with ribonuclease and subsequent dialysis. Deoxyribonucleic acid was reprecipitated with 2-ethoxyethanol. The final product contained less than 1% protein. The deoxyribonucleic acid obtained from
S. aureus
strain S
44
had a (adenosine + thymine)/(guanine + cytosine) base ratio of 1.98. Intrinsic viscosity in 0.15
m
NaCl + 0.015
m
sodium citrate was approximately 76 dl/g. The sedimentation coefficient,
S
20
w, was close to 25 S.
NEW SOLID MEDIUM FOR ENHANCED GROWTH OF
PASTEURELLA TULARENSIS
Authors: Gaspar Andrew J. , Tresselt Hugh B. , Ward Martha K.
Published: January 2020

Gaspar, Andrew
J. (Fort Detrick, Frederick, Md.),
Hugh B. Tresselt, and Martha K. Ward
. New solid medium for enhanced growth of
Pasteurella tularensis
. J. Bacteriol.
82:
564–569. 1961.—The purpose of this work was to develop a solid medium for more rapid growth of
Pasteurella tularensis
, especially from small inocula comparable to those generally encountered in clinical materials.

After titration of various ingredients, separately and in combination, the optimal base was found to be: 2.6% tryptose broth (Difco) with thiamine, 0.5% cysteine-HCl, 0.2% sodium thioglycolate, 1.0% glucose, dissolved by mixing without heat, and adjusted to pH 7.2 ± 0.03. To this base 1.0% agar (Difco) is added and dissolved by heating in flowing steam for about 5 min prior to autoclaving at 121 C for 20 min. After sterilization and cooling, 5.0% defibrinated rabbit blood is added aseptically. Plates of the completed medium are incubated at 37 C for 24 hr prior to use.

Colonies of
P. tularensis
approximately 1.0 mm in diameter are obtained on this medium after 26 to 29 hr incubation if conditions of high relative humidity (90 to 100% saturation) are maintained.

The mechanisms involved in the growth enhancement obtained on this medium are under study. The role of thioglycolate appears to be that of keeping in solution the relatively high concentration of cysteine-HCl required. The specific methods of preparation described, the incubation of plates prior to use, and final incubation under conditions of high humidity all are important for optimal results.

This developmental work has employed only pure cultures. Attempts are being made to develop a selective medium, using this new preparation as base, for the direct isolation of
P. tularensis
from a variety of clinical materials.
UTILIZATION OF PYRIMIDINES AND PYRIMIDINE DEOXYNUCLEOSIDES BY
THERMOBACTERIUM ACIDOPHILUM
(
LACTOBACILLUS ACIDOPHILUS
)
Authors: Løvtrup Søren , Shugar David
Published: January 2020

Løvtrup, Søren
(University of Göteborg, Sweden)
and David Shugar
. Utilization of pyrimidines and pyrimidine deoxynucleosides by
Thermobacterium acidophilum
(
Lactobacillus acid-ophilus
). J. Bacteriol.
82:
623–631. 1961.—The utilization of pyrimidine deoxynucleosides was investigated by means of deoxyribosides of unnatural pyrimidines, especially by halogen-substituted uracil derivatives. All investigated deoxyribosides could be used, except that of
N
-methylthymidine. It was concluded that this substance cannot be a substrate for the enzyme
trans-N
-deoxyribosylase, which has been shown to be active in the utilization of deoxyribosides in this microorganism. With uracil as the only pyrimidine source, the halogen-substituted deoxyuridines had a certain inhibitory effect on growth.

Contrary to previous findings, it was observed that normal growth occurs in the presence of thymine as the only pyrimidine source. The utilization of this substance is less efficient than that of uracil; a 1:10 dilution leads to a decrease in the extent of growth with the former, but not with the latter. From these results, complemented with experiments in which halogen-substituted uracil derivatives and the corresponding ribosides or deoxyribosides were used as inhibitors, it has been possible to account for most of the metabolic interconversions of pyrimidines in the investigated microorganism.
STAPHYLOCOCCAL MULLER PHENOMENON: RELATIONSHIP TO THE PLASMINOGEN-PLASMIN SYSTEM
Authors: Quie Paul G. , Wannamaker Lewis W.
Published: January 2020

Quie, Paul
G. (University of Minnesota, Minneapolis)
and Lewis W. Wannamaker
. Staphylococcal Muller phenomenon: relationship to the plasminogen-plasmin system. J. Bacteriol.
82:
770–783. 1961.—The staphylococcal factor that produces particulate proteolysis (Muller phenomenon) in whole blood or hemoglobinserum agar plates has been obtained in supernatants from broth cultures. This has facilitated study of the phenomenon in a sterile system. The bacterial factor has been partially separated from other extracellular products by starch zone electrophoresis.

Staphylococcal Muller factor and staphylokinase are similar in heat stability, in electrophoretic mobility, and in activation of a serum proteolytic system. A relationship of the Muller phenomenon to the plasminogen-plasmin system is also suggested by the observation that purified plasminogen may be substituted for serum in the sterile system, and by the finding that inhibitors of the plasminogen-plasmin system also inhibit the Muller phenomenon. Although growing colonies of streptococci fail to exhibit the Muller phenomenon, particulate proteolysis can be demonstrated with streptokinase, using a double-diffusion technique. A similar phenomenon can be produced with urine, suggesting that urokinase may substitute for the bacterial activators in the Muller phenomenon.

Although evidence from other sources suggests that the serum Muller factor is particulate, the serum factor appears to be freely diffusible in agar and is not readily sedimented in the ultracentrifuge.
SIALIC ACIDS (
N
,7-O-DIACETYLNEURAMINIC ACID AND
N
-ACETYLNEURAMINIC ACID) IN
ESCHERICHIA COLI
II
Authors: Dewitt Charles W. , Zell Elizabeth A.
Published: January 2020

DeWitt, Charles
W. (The Upjohn Co., Kalamazoo, Mich.)
and Elizabeth A. Zell
. Sialic acids (
N
,7-O-diacetylneuraminic acid and
N
-acetylneuraminic acid) in
Escherichia coli
. II. Their presence on the cell wall surface and relationship to K antigen. J. Bacteriol.
82:
849–856. 1961.—Sialic acids obtained from
Escherichia coli
by weak acid hydrolysis are shown to be released from the extreme-surface, lipoprotein layer of the cell. Hydrolysis does not lyse the cells nor release cell walls. Hydrolysates do not contain compounds such as α, ε-diaminopimelic acid or
N
-acetylmuramic acid, normally associated with cell walls. The surface material is released in a relatively homogeneous form by simple incubation of live cells in an acidic buffer and is susceptible to hydrolysis by viral neuraminidase, although untreated live cells will not serve as substrate. As this surface material resembles, in its initial rate of reaction with neuraminidase, human urinary mucoprotein, the sialic acid is presumably present as an α-ketoside. Colominic acid is insusceptible under the same conditions. Although there is good correlation of the occurrence of sialic acids with the
E. coli
capsular antigen, K
1
, specific release of sialic acid from K
1
antigen with neuraminidase does not lessen its haptenic activity as measured by inhibition of specific agglutination.
METABOLIC CHARACTERIZATION OF THE GENUS
BRUCELLA
IV. 3
Authors: Meyer Margaret E.
Published: January 2020

Meyer, Margaret
E. (University of California, Davis). Metabolic characterization of the genus
Brucella
. IV. Correlation of oxidative metabolic patterns and susceptibility to
Brucella
bacteriophage, type
abortus
, strain 3. J. Bacteriol.
82:
950–953. 1961.—A total of 212 strains of brucellae that had been identified as
Brucella melitensis, B. abortus, B. suis, or B. neotomae
by their oxidative metabolism were tested for their susceptibility to
Brucella
bacteriophage, type
abortus
, strain 3. It was demonstrated that only those organisms that displayed the oxidative metabolic pattern that is singular for
B. abortus
were susceptible to this strain of phage, irrespective of their identity by the conventional methods usually employed for differentiating members of this genus. Strains of organisms that display the features of
B. melitensis
by the conventional determinative methods, but display the metabolic characteristics of
B. abortus
, are susceptible to lysis by this phage. These organisms are in fact
B. abortus
. Strains of organisms that display the features of
B. melitensis
by the classical methods, and display the metabolic pattern of
B. melitensis
, are not lysed by this phage. These organisms are
B. melitensis
. The conclusions then were drawn that
B. abortus
is the only species that can serve as host for this strain of phage, that oxidative metabolic patterns accurately identify the species in this genus, and that by the conventional methods of differentiation, many strains of
B. abortus
are misidentified as
B. melitensis
.
INFLUENCE OF GUINEA PIG PLASMA FACTORS ON PHAGOCYTOSIS OF
PASTEURELLA PESTIS
Authors: Stanziale W. G. , White J. D.
Published: January 2020

Stanziale, W. G.
(Fort Detrick, Frederick, Md.)
and J. D. White
. Influence of guinea pig plasma factors on phagocytosis of
Pasteurella pestis
. II. Plasma from plague-infected guinea pigs. J. Bacteriol.
83:
182–186. 1962.—The phagocytosis enhancing property of normal guinea pig plasma was altered during experimental plague infection. The most notable changes occurred in the plasma from afebrile, moribund animals and from those convalescing from acute infection. The plasma of the moribund guinea pigs, which was obtained 7 to 8 days after exposure to
Pasteurella pestis
, inhibited phagocytosis to a considerable degree. Plasma from convalescent guinea pigs enhanced phagocytosis to a higher degree than the plasma of normal animals. In contrast, plasma from recovered guinea pigs in which cultural or serological evidence of infection was not demonstrated enhanced phagocytosis to a degree equivalent to that of normal plasma. Plasma taken from guinea pigs during the febrile phase of plague infection enhanced phagocytosis to a lower degree than did normal plasma.
TRYPTOPHAN SYNTHETASE LEVELS IN
ESCHERICHIA COLI, SHIGELLA DYSENTERIAE
, AND TRANSDUCTION HYBRIDS
Authors: Eisenstein Richard B. , Yanofsky Charles
Published: January 2020

Eisenstein, Richard B.
(Western Reserve University, Cleveland, Ohio)
and Charles Yanofsky
. Tryptophan synthetase levels in
Escherichia coli, Shigella dysenteriae
, and transduction hybrids. J. Bacteriol.
83:
193–204. 1962—
Shigella dysenteriae
and
Escherichia coli
, strains K-12 and B, were found to produce low levels of tryptophan synthetase, although some hybrids, formed by the introduction of the gene cluster concerned with tryptophan synthesis from
S. dysenteriae
into
E. coli
, produced high levels of this enzyme system. A revertant obtained from a tryptophan-requiring mutant also formed high levels of tryptophan synthetase. The gene or genes responsible for high enzyme production in these strains was shown to be linked to the cluster of genes concerned with tryptophan synthesis. The cause of high enzyme production was investigated. Various lines of evidence, including stimulation of growth by tryptophan precursors, sensitivity to inhibition by 5-methyltryptophan, absence of accumulation of tryptophan, and repression of enzyme formation by anthranilic acid and tryptophan, suggested that high enzyme production in the strains examined results from a partial block in the tryptophan pathway and not from resistance to repression by tryptophan. The conversion of shikimic acid-5-phosphate to anthranilic acid appears to be the partially blocked reaction in the strains studied.
PERMEABILITY OF BACTERIAL SPORES III
Authors: Black S. H. , Gerhardt Philipp
Published: January 2020

Black
, S. H. (The University of Michigan, Ann Arbor)
and Philipp Gerhardt
. Permeability of bacterial spores. III. Permeation relative to germination. J. Bacteriol.
83:
301–308. 1962.—The passive diffusion of solutes into dormant spores, characterized previously with the test organism
Bacillus cereus
strain terminalis, has now been examined in relation to germination. Dormant spores did not take up specific germinants differently than they did other compounds, under conditions optimal for germination. Germinated spores, viable but prevented from growing out, displayed some changes in permeability, evidenced by increased total uptake of glucose and water and by observable penetration of a fluorescigenic dye. Heat-killed spores were as permeable to glucose and the dye as germinated ones.
EFFECT OF
p
-FLUOROPHENYLALANINE ON PSITTACOSIS VIRUS IN TISSUE CULTURES
Authors: Tanami Yoh , Pollard Morris
Published: January 2020

Tanami, Yoh
(University of Texas Medical Branch, Galveston)
and Morris Pollard
. Effect of
p
-fluorophenylalanine on psittacosis virus in tissue cultures. J. Bacteriol.
83:
437–442. 1962.—The inhibitory effect of
p
-fluorophenylalanine (FPA) on maturation of psittacosis virus was investigated, with attention to the time sequence of viral protein synthesis. Extracellular virus particles were not inactivated by FPA at a concentration of 100 μg per ml, at which level it interfered with maturation of intracellular virus. When FPA was added to infected tissue cultures earlier than 15 hr after infection, intracellular virus maturation was suppressed. However, when FPA was added after 15 hr, infective virus was produced, which indicates that the synthesis of a FPA-sensitive virus precursor (presumably viral protein) had already occurred. A latent (“dormant”) infection of psittacosis virus, established in a medium deficient in phenylalanine and tyrosine, was also investigated.
FLUORESCENT ANTIBODY STAINING AND AGGLUTINATION REACTIONS IN
BORDETELLA PERTUSSIS
CULTURES
Authors: Eldering Grace , Eveland Warren C. , Kendrick Pearl L.
Published: January 2020

Eldering, Grace
(Michigan Department of Health, Grand Rapids),
Warren C. Eveland, and Pearl L. Kendrick
. Fluorescent antibody staining and agglutination reactions in
Bordetella pertussis
cultures. J. Bacteriol.
83:
745–749. 1962—
Bordetella pertussis
antisera produced with smooth cultures gave positive results in agglutination and fluorescent antibody (FA) tests with smooth
B. pertussis
cultures. Similar results were obtained with antisera produced with the related species
B. parapertussis
and
B. bronchiseptica
when each was tested with the respective homologous antigen. Cross reactions occurred with some of the antisera and heterologous antigens but a positive agglutination test was not always correlated with a positive FA reaction, and the reverse was also observed. Adsorbed
B. pertussis
antisera specific for factors 2, 3, 4, and 5 agglutinated appropriate
B. pertussis
cultures, but gave negative FA reactions. Factor 1 antiserum prepared by adsorption with heated (100 C) antigen gave positive results in both FA and agglutination tests. Rough
B. pertussis
cultures were inagglutinable, but were FA positive when tested with unadsorbed antiserum produced with either smooth or rough cultures. Rough cultures were FA negative with factor 1 serum. The results suggest that for
B. pertussis
different serum components may be responsible for agglutination and FA staining. The implications with respect to the protection-inducing antigen are discussed.
MUTATION TO NEOSTIGMINE RESISTANCE IN A CHOLINESTERASE-CONTAINING
PSEUDOMONAS
Authors: Searle Barbara W. , Goldstein Avram
Published: January 2020

Searle, Barbara
W. (Stanford University School of Medicine, Palo Alto, Calif.)
and Avram Goldstein
. Mutation to neostigmine resistance in a cholinesterase-containing
Pseudomonas
. J. Bacteriol.
83:
789–796. 1962.—In a strain of
Pseudomonas fluorescens
containing an inducible cholinesterase, the activity of that enzyme is rate-limiting for growth when acetyl-choline is the sole source of carbon or nitrogen. Under these circumstances, neostigmine, a cholinesterase inhibitor, becomes a growth inhibitor.

A neostigmine-resistant mutant was isolated, and the properties of its cholinesterase were compared with those of the wild-type enzyme. There were no differences in penetration of cells by inhibitor, rate of enzyme-inhibitor combination, affinity of inhibitor or substrate for the cholinesterase, or protective effect of substrate upon the enzyme. However, the mutant consistently formed cholinesterase at about twice the wild-type rate. Mutation, in this case, appears to result in a specific change in the differential rate of enzyme biosynthesis. The relationship of this change to neostigmine resistance is discussed, and it is suggested that the effect observed here may be prototypic of a general type of mechanism responsible for acquired drug resistance.
PEPTIDES REQUIRED FOR THE GROWTH OF
LACTOBACILLUS BULGARICUS
Authors: Jones K. M. , Woolley D. W.
Published: January 2020

Jones
, K. M. (The Rockefeller Institute, New York, N. Y.)
and D. W. Woolley
. Peptides required for the growth of
Lactobacillus bulgaricus
. J. Bacteriol.
83:
797–801. 1962.—
Lactobacillus bulgaricus
09 had an absolute requirement for growth for a suitable peptide, which was satisfied by trypsinized casein or hemoglobin. Digests of globin and the α and β chains derived from it showed similar activity to hemoglobin digests. Each of four highly purified peptides of different amino acid composition isolated from the α chain of globin had activity. Each active peptide had seven or more residues; smaller peptides were inactive. The requirement for
L. bulgaricus
was not one for strepogenin because some substances with strepogenin activity for
L. casei
were not active for
L. bulgaricus
.
METABOLIC FUNCTION OF BRANCHED-CHAIN VOLATILE FATTY ACIDS, GROWTH FACTORS FOR RUMINOCOCCI II
Authors: Allison M. J. , Bryant M. P. , Katz I. , Keeney M.
Published: January 2020

Allison
, M. J. (Dairy Cattle Research Branch, USDA, Beltsville, Md.), M. P.
Bryant, I. Katz, and M. Keeney
. Metabolic function of branched-chain volatile fatty acids, growth factors for ruminococci. II. Biosynthesis of higher branched-chain fatty acids and aldehydes. J. Bacteriol.
83:
1084–1093. 1962.—A number of strains of rumen bacteria require branched-chain volatile fatty acids for growth. A strain of
Ruminococcus flavefaciens
that requires either isovalerate or isobutyrate incorporates radioactive carbon from isovalerate-1-C
14
and isovalerate-3-C
14
into leucine and into the lipid fraction of the cells. Evidence obtained by both paper and gas chromatography indicated that most of the label in the lipid of cells grown in isovalerate-1-C
14
was in a branched-chain 15-carbon fatty acid, with some in a 17-carbon acid; about 7.5% of the C
14
was recovered in a branched-chain 15-carbon aldehyde. The aldehydes were in the phospholipid fraction and were presumably present as plasmalogen.

A strain of
R. albus
was shown to require isobutyrate, 2-methyl-
n
-butyrate, or 2-ketoisovalerate for growth. This strain did not incorporate appreciable C
14
from isovalerate-1-C
14
or isovalerate-3-C
14
. When grown in a medium containing isobutyrate-1-C
14
, most of the cellular C
14
was found in the lipid fraction. Analysis of the lipid demonstrated that the label was present mainly as branched-chain 14-carbon and 16-carbon fatty acids, with 11% of the C
14
present in 14- and 16-carbon carbonyl compounds, presumably branched-chain aldehydes.

Branched-chain 14-, 15-, and 16-carbon fatty acids are major components of the lipids of these rumen bacteria. The possibility that these acids and aldehydes, which are found in ruminant body and milk lipids, may be of microbial origin is discussed.
EFFECTS OF NEAR-ULTRAVIOLET IRRADIATION ON GROWTH AND OXIDATIVE METABOLISM OF BACTERIA
Authors: Kashket E. R. , Brodie A. F.
Published: January 2020

Kashket
, E. R. (Harvard Medical School, Boston, Mass.)
and A. F. Brodie
. Effects of near-ultraviolet irradiation on growth and oxidative metabolism of bacteria. J. Bacteriol.
83:
1094–1100. 1962.—The effects of irradiation with near-ultraviolet light (360 mμ) have been studied with
Escherichia coli
W and a strain of
Pseudomonas aeruginosa
. The growth of the aerobe
P. aeruginosa
was inhibited by light on minimal salts media containing succinate, glutamate, or glucose as sole carbon sources. The facultative anaerobe
E. coli
was capable of growth under irradiation on a fermentable carbon source, such as glucose, but with a smaller yield of cells on limiting substrate, as compared to unirradiated control cultures. The rate of growth of aerobic irradiated cells on glucose was equal to that of anaerobic growth on that carbon source, and there was a greater accumulation of end products of glucose catabolism aerobically in the light as compared to dark controls. When irradiated in media containing carbon sources from which energy was obtainable only by oxidative phosphorylation, such as succinate or malate,
E. coli
cells were still capable of oxidizing these substrates but could not grow on them. This bacteriostatic effect of 360-mμ light could be reversed by the addition of glucose, which resulted in the growth of irradiated cells. Visible (400 to 600 mμ) light was found to have no effect. Irradiated
E. coli
cells in succinate were found to contain no naphtho- or benzoquinones, compounds which are more sensitive to 360-mμ irradiation than other components of the respiratory chain. It is suggested that the effect of 360-mμ light on whole cells is the destruction of light-sensitive components, such as the benzoquinone Q
8
and naphthoquinone K
2
C
45
of
E. coli
W, which are essential for obtaining energy from oxidative metabolism.
PYRITHIAMINE ADAPTATION OF
STAPHYLOCOCCUS AUREUS
I
Authors: Das S. K. , Chatterjee G. C.
Published: January 2020

Das
, S. K. (University of Calcutta, Calcutta, India)
and G. C. Chatterjee
. Pyrithiamine adaptation of
Staphylococcus aureus
. I. Adaptation and carbohydrate utilization. J. Bacteriol.
83:
1251–1259. 1962.—
Staphylococcus aureus
has been adapted to pyrithiamine, a thiamine analogue; as a result of this adaptation, the color of the pigment of the organism changes from orange-yellow to lemon-yellow. The adaptation is reversible; the adapted strain will revert after repeated subculture in a medium containing thiamine and no pyrithiamine. Of the major biochemical alterations resulting from adaptation, severe depression in glucose utilization and simultaneous stimulation of acetate utilization have been noticed. The effect of metabolic inhibitors on the utilization of glucose and acetate has also been studied. By measuring the rate of formation of C
14
O
2
from glucose-1-C
14
and glucose-6-C
14
, it has been observed that the reduction in C
14
O
2
formation from glucose-1-C
14
by the adapted organism is much more than that obtained from glucose-6-C
14
, causing thereby a decreased metabolic ratio of these two substrates after such adaptation. Relative to the normal strain, the adapted strain utilizes acetate-C
14
at a much faster rate, both in the formation of C
14
O
2
and also in the incorporation of C
14
into the protein and lipid fractions; the rate of formation of C
14
O
2
from pyruvate-1-C
14
is not greatly altered. It has been postulated that there is a partial blocking of the pentose phosphate cycle, because of the lowered glucose-1-C
14
utilization, and simultaneous stimulation of the tricarboxylic acid cycle; or perhaps the initiation of some other route after pyrithiamine adaptation would account for the great increase in acetate utilization.
REQUIREMENT OF DIVALENT METAL IONS FOR BACITRACIN ACTIVITY
Authors: Adler Richard H. , Snoke John E.
Published: January 2020

Adler, Richard
H. (University of California, Los Angeles)
and John E. Snoke
. Requirement of divalent metal ions for bacitracin activity. J. Bacteriol.
83:
1315–1317. 1962.—The antibiotic activity of bacitracin towards
Staphylococcus aureus
is completely suppressed by ethylenediaminetetraacetate. The conclusion that bacitracin activity requires the presence of metal ions is supported by the demonstration that the addition of a number of divalent ions reverses the effect of ethylenediaminetetraacetate. The most active of these ions, cadmium, manganese, and zinc, directly enhance the activity of bacitracin.
l
-ARABINOSE-SENSITIVE,
l
-RIBULOSE 5-PHOSPHATE 4-EPIMERASE-DEFICIENT MUTANTS OF
ESCHERICHIA COLI
Authors: Englesberg E. , Anderson R. L. , Weinberg R. , Lee N. , Hoffee P , Huttenhauer G. , Boyer H.
Published: January 2020

Englesberg
, E. (University of Pittsburgh, Pittsburgh, Pa.),
R L. Anderson, R. Weinberg, N. Lee, P. Hoffee, G. Huttenhauer, and H. Boyer
.
l
-Arabinose-sensitive,
l
-ribulose 5-phosphate 4-epimerase-deficient mutants of
Escherichia coli
. J. Bacteriol.
84:
137–146. 1962—
l
-Arabinose-negative mutants of
Escherichia coli
B/r, ara-53 and ara-139, are deficient in the enzyme
l
-ribulose 5-phosphate 4-epimerase; ara-53, further analyzed, accumulates large quantities of
l
-ribulose 5-phosphate when incubated with
l
-arabinose. The mutant sites are closely linked to the left of the previously ordered
l
-arabinose mutant sites, and probably represent the structural gene for
l
-ribulose 5-phosphate 4-epimerase (gene D) in the
l
-arabinose operon. The inducible levels of
l
-arabinose isomerase and
l
-ribulose 5-phosphate 4-epimerase vary correspondingly as a result of mutation in the structural gene for
l
-ribulokinase (gene B), further substantiating the dual structural and regulatory function of this gene locus. Ara-53 and ara-139 are strongly inhibited by
l
-arabinose and give rise to
l
-arabinose-resistant mutants. The one resistant mutant analyzed still lacks the 4-epimerase but is deficient in
l
-ribulokinase and has increased
l
-arabinose isomerase activity, a characteristic of a type of mutation in the B gene. It is proposed that accumulation of
l
-ribulose 5-phosphate is responsible for the inhibition, and that mutation to resistance will involve mutation in the A, B, C, permease, or repressor genes, thus providing a direct method for isolating these types of
l
-arabinose-negative mutants. Glucose prevents and cures the
l
-arabinose inhibition.
MICROBIOLOGICAL ASSAY FOR THYMINE USING A MUTANT OF
STREPTOCOCCUS FAECALIS
Authors: Doctor V. M.
Published: January 2020

Doctor
, V. M. (University of Texas, Houston). Microbiological assay for thymine using a mutant of
Streptococcus faecalis
. J. Bacteriol.
84:
17–22. 1962.—A rapid microbiological assay procedure for the determination of thymine was devised. The test organism,
Streptococcus faecalis
Akt, a mutant of
S. faecalis
ATCC 8043, gives one-half the maximal growth response to 1.6 μg of thymine (assay range 0.5 to 2.5 μg) or to a molar equivalent (3.2 μg) of thymidine. Under these experimental conditions, 80 μg of 5-methylcytosine or 150 μg of thymidylic acid replace the thymine or thymidine requirement of the test organism. Addition of a mixture of adenine, guanine, and xanthine at a level of 40 μg/ml of each or higher to the basal medium, or addition of a mixture of deoxyadenosine and deoxyguanosine at a level of 80 μg/ml of each or higher to the basal medium, inhibited the thymine or thymidine growth response of the organism. Thymine activities of rat and chick tissues were determined by this procedure. The data on the recovery of added thymine to tissues gave 84 to 103% recovery. Thymus, spleen, small intestine, and ventral prostate were higher in thymine activities as compared with other tissues.
EVALUATION OF EXTRACELLULAR DEOXYRIBONUCLEASE ACTIVITY INPSEUDOMONAS
Authors: Streitfeld Murray M. , Hoffmann Edward M. , Janklow Herbert M.
Published: January 2020

Streitfeld, MurrayM. (University of Miami, Coral Gables, Fla.),Edward M. Hoffmann, and Herbert M. Janklow. Evaluation of extracellular deoxyribonuclease activity inPseudomonas. J. Bacteriol.84:77–80. 1962.—Of 23 strains ofP. aeruginosaisolated from pathological processes, 22 strains, representative of at least nine different somatic antigen types, were found to elaborate extracellular deoxyribonuclease. Qualitative determinations of enzyme elaboration were performed after the development of a convenient toluidine-blue assay plate procedure, dependent upon the metachromatic properties of the dye. This new procedure permits the determination of enzyme activity despite the presence, in the deoxyribonucleic acid-containing agar medium, of 2 to 5 ml/100 ml serum or plasma, or extracellular pyocyanin.