bluuebunny/crossref_metadata_2025 huggingface dataset:
Nutritional requirements of anaerobic coryneforms
Authors: Ferguson D A , Cummins C S
Published: January 2020

The nutritional requirements of three species of anaerobic coryneforms and their serotypes (Propionibacterium acnes types I and II, P. avidum types I and II, and P. granulosum) were determined. Strains of P. avidum would consistently grow to a transmittance of 1 to 3% at 560 nm in a basal salts medium supplemented with glucose, pantothenate, biotin, thiamine, and 12 amino acids (alanine, arginine, cysteine, glutamine, glycine, histidine, isoleucine, methionine, phenylalanine, serine, tyrosine, and tryptophan). Strains of P. acnes and P. granulosum, however, failed to grow in this medium unless six additional amino acids were present (asparagine, leucine, lysine, proline, threonine, and valine). All three species grew equally well whether the 18 amino acids were supplied in the form of a casein hydrolysate supplemented with tryptophan or were added separately. Nicotinamide enhanced growth of P. acnes but had no effect on growth of P. avidum and P. granulosum. Other nutrients which were not absolute requirements, but which significantly improved growth of these species, included the purines guanine and/or adenine, Tween 80, which served as a source of oleic acid, sodium L-lactate, alpha-ketoglutarate, and pyruvate. Strains (86) comprising all five groups grew well in the defined medium, except four strains of P. acnes type II (29 tested), which failed to grow unless heme and vitamin K were added to the medium. One strain of P. granulosum (22 tested) failed to grow in any defined medium, suggesting an additional growth factor requirement.
Aspartate Transcarbamylase Synthesis Ceases Prior to Inactivation of the Enzyme in
Bacillus subtilis
Authors: Maurizi Michael R. , Switzer Robert L.
Published: January 2020

Aspartate transcarbamylase is synthesized during exponential growth of
Bacillus subtilis
and is inactivated when the cells enter the stationary phase. This work is a study of the regulation of aspartate transcarbamylase synthesis during growth and the stationary phase. Using specific immunoprecipitation of aspartate transcarbamylase from extracts of cells pulse-labeled with tritiated leucine, we showed that the synthesis of the enzyme decreased very rapidly at the end of exponential growth and was barely detectable during inactivation of the enzyme. Synthesis of most cell proteins continued during this time. When the cells ceased growing because of pyrimidine starvation of a uracil auxotroph, however, synthesis and inactivation occurred simultaneously. Measurement of pools of pyrimidine nucleotides and guanosine tetra- and pentaphosphate demonstrated that failure to synthesize aspartate transcarbamylase in the stationary phase was not explained by simple repression by these compounds. The cessation of aspartate transcarbamylase synthesis may reflect the shutting off of a “vegetative gene” as part of the program of differential gene expression during sporulation. However, aspartate transcarbamylase synthesis decreased normally at the end of exponential growth at the nonpermissive temperature in a mutant strain that is temperature-sensitive in sporulation and RNA polymerase function. Cessation of aspartate transcarbamylase synthesis appeared to be normal in three other temperature-sensitive RNA polymerase mutants and in several classes of
spo0
mutants.
Synchronous division and rates of macromolecular synthesis in Haemophilus influenzae competent for genetic transformation
Authors: Scocca J J , Habersat M
Published: January 2020

When competent Haemophilus influenzae were returned to complete growth medium, they resumed growth at once and underwent a synchronous division after 20 to 25 min. The rate of DNA synthesis increased to the definitive rate very quickly after resumption of growth. In contrast, the rate of RNA synthesis remained low until the time of division, at which time it accelerated dramatically. Changes in the rate of protein synthesis were similar in form to those of RNA synthesis. The kinetics of loss of competence upon resumption of growth had a pattern similar to the rate of RNA synthesis, and both processes accelerated markedly at the time of cell division, suggesting an inverse relationship between RNA synthesis and the ability to take up DNA.
Role of the Escherichia coli aromatic amino acid aminotransferase in leucine biosynthesis
Authors: Powell J T , Morrison J F
Published: January 2020

Strains of Escherichia coli that lack the branched-chain amino acid amino-transferase because of mutations in the ilvE gene had no growth requirement for leucine when the cells contained the aromatic amino acid aminotransferase that is the product of the tyrB gene. The presence of leucine increased the generation time of these cells and decreased the specific activity of the aromatic amino acid aminotransferase. It is concluded that this enzyme functions efficiently in leucine biosynthesis and can be repressed by leucine as well as by tyrosine.
Phospholipid composition of Rickettsia prowazeki grown in chicken embryo yolk sacs
Authors: Winkler H H , Miller E T
Published: January 2020

The phospholipid composition and phospholipid fatty acid composition of purified Rickettsia prowazeki were determined. The lipid phosphorous content was 6.8 +/- 1.3 microgram/mg of total rickettsial protein. The major phospholipid was phosphatidylethanolamine (60 to 70%); phosphatidylglycerol constituted 20%, and phosphatidylcholine constituted 15%. Small amounts of phosphatidylserine and cardiolipin were detected. The principal fatty acids were 18:1, 16:1, and 16:0. The fatty acid composition of the phosphatidylcholine in the rickettsial extracts was very different than that of the other rickettsial phosphatides and very similar to that of normal yolk sac phosphatidylcholine. The specific of the phosphatidylcholine of rickettsiae grown in the presence of 32P was markedly lower than that of phosphatidylethanolamine and phosphatidylglycerol. It is suggested that the phosphatidylcholine in the rickettsial extract is yolk sac derived and either tightly absorbed or exchanged into the rickettsial membrane.
Kinetics of accumulation of a photodynamically induced cell-surface polypeptide in a species of Arthrobacter
Authors: Hoober J K
Published: January 2020

Cells of a species of Arthrobacter were incubated in the light with methylene blue, a dye that sensitizes photooxidative reactions by the production of singlet oxygen. An early and major response by the cells to these conditions was stimulation of synthesis of a single cell-surface polypeptide, 21,000 daltons in mass. The rate of synthesis of this polypeptide reached a maximal level about 30 min after the start of illumination. As a consequence, the amount of this polypeptide increased at least 10-fold during a period of 5 h. The presence of histidine or methionine, scavengers of singlet oxygen, markedly diminished synthesis and accumulation of this polypeptide. Concomitant with the accumulation of this polypeptide on the cell surface was the appearance of an extensive array of pili.
Repair of cross-linked DNA and survival of Escherichia coli treated with psoralen and light: effects of mutations influencing genetic recombination and DNA metabolism
Authors: Sinden R R , Cole R S
Published: January 2020

Repair of cross-linked DNA was studied in Escherichia coli strains carrying mutations affecting DNA metabolism. In wild-type cells, DNA strands cut during cross-link removal were rejoined during a subsequent incubation into high-molecular-weight molecules. This rejoining was dependent on gene products involved in genetic recombination. A close correlation was found relating recombination proficiency, the rate of strand rejoining, and formation of viable progeny after DNA cross-linking by treatment with psoralen and light. Wild-type cells and other mutants which were Rec+ (sbcB, recL, recL sbcB, recB recC sbcA, recB recC sbcB, xthA1, and xthA11) rejoined cut DNA strands at a rate of 0.8 +/- 0.1 min -1 at 37 degrees C and survived 53 to 71 cross-links per chromosome. recB, recC, recB recC, recF, or polA strains showed reduced rates of strand rejoining and survived 4 to 13 cross-links per chromosome. Recombination-deficient strains (recA, recB recC sbcB recF, recB recL) and lexA failed to rejoin DNA strands after crosslink removal and were unable to form colonies after treatments producing as few as one to two cross-links per chromosome. Strand rejoining occurred normally in cells with mutations affecting DNA replication (dnaA, danB, dnaG, and dnaE) under both permissive and nonpermissive conditions for chromosome replication. In a polA polB dnaE strain strand rejoining occurred at 32 degree C but not at 42 degree C, indicating that some DNA synthesis was required for formation of intact recombinant molecules.
Synthesis and salvage of purines during cellular morphogenesis of Myxococcus xanthus
Authors: Tsai W C , Westby C A
Published: January 2020

Intact cells of Myxococcus xanthus were examined for de novo purine synthesis and salvage utilization. The cellular uptake rates of radioactive glycine (de novo purine precursor), adenine, and guanine were measured, and thin-layer chromatography and radioautography were used to examine cell extracts for de novo synthesized purine nucleotides. Intact vegatative cells, glycerol-induced myxospores, and germinating cells of M. xanthus CW-1 were able to carry out de novo purine and salvage synthesis. Germinating cells and glycerol-induced myxospores were metabolically more active or as active as vegetative cells with respect to purine anabolism. We conclude that M. xanthus is capable of synthesizing purine nucleotides and salvaging purines throughout the glycerol version of its life cycle.
Occurrence of squalene and sterols in Cellulomonas dehydrogenans (Arnaudi 1942) comb. nov. Hester 1971
Authors: Weeks O B , Francesconi M D
Published: January 2020

The neutral lipid fraction of the photochromogenic, coryneform bacterium Cellulomonas dehydrogenans (Arnaudi 1942) comb. nov. contains the sterol precursor squalene and at least two sterols, cholesterol and beta-sitosterol. The compounds were characterized by mass spectrometry and combination gas-liquid chromatography--mass spectrometry. De novo sterol biosynthetic ability was shown from incorporation of 14C from D-[U-14C]glucose into squalene and the sterol fraction. The squalene concentration approximated 0.002 to 0.005% of the total dry cell weight, and the sterols approximated 0.03 to 0.05%.
Role of lipopolysaccharide and outer membrane protein of Escherichia coli K-12 in the receptor activity for bacteriophage T4
Authors: Mutoh N , Furukawa H , Mizushima S
Published: January 2020

Lipopolysaccharide isolated from Escherichia coli K-12 did not inactivate phage T4, although the cell envelopes with 1% sodium deoxycholate resulted in the release of cytoplasmic membrane proteins, 70% of the lipopolysaccharide, and almost all of the phospholipid. The reconstitution of phage receptor activity was achieved from deoxycholate-soluble and -insoluble fractions by dialysis against a solution of magnesium chloride. Lipopolysaccharide was the only essential component in the deoxycholate-soluble fraction. PhageT4-resistant mutants YA21-6 and YA21-82, having defects in the deoxycholate-soluble and -insoluble fractions, respectively, were isolated. The deoxycholate-soluble fraction of YA21-6 possessed heptoseless lipopolysaccharide, and this defect was responsible for the phage resistance. The deoxycholate-insoluble fraction of YA21-82 lacked outer membrane protein O-8. The addition of O-8 to this fraction together with the wild-type lipopolysaccharide resulted in the appearance of the receptor activity. Furthermore, the reconstitution was successfully achieved with only O-8 and the wild-type lipopolysaccharide, indicating that O-8 was an essential component in the deoxycholate-insoluble fraction.
Plasmids of incompatibility group P code for the capacity to propagate bacteriophage IKe
Authors: Grant R B , Whiteley M H , Shapley A J
Published: January 2020

Seven of eight plasmids of incompatibility group P were found to code for the capacity to propagate bacteriophage IKe in Escherichia coli. Six of the seven plasmids allowed propagation of IKe by one bacterial host (RG172) but not by another (RG176); the other plasmid allowed IKe propagation by both hosts. IKe propagation by a number of E. coli K-12 strains was quite variable. IKeh, an extended host range mutant of IKe, was found to plaque specifically on N+ and P+ strains.
Isolation and characterization of a membrane-bound, low-potential c-type cytochrome from purple photosynthetic bacteria, with special reference to Rhodospirillum rubrum
Authors: Yoch D C , Carithers R P , Arnon D I
Published: January 2020

Other investigators have isolated soluble, low-potential, c-type cytochromes (cytochrome c3) from a few photosynthetic procaryotes, i.e., a cyanobacterium and two species of purple nonsulfur bacteria. However, such cytochromes appeared to be absent from other purple bacteria, including Rhodospirillum rubrum and Chromatium vinosum. We now report evidence for the presence of low-potential c-type cytochromes in these two species, in which they were found to be bound to the photosynthetic membranes. Evidence for a membrane-bound, low-potential c-type cytochrome was also found in Rhodopseudomonas sphaeoides. The low-potential c-type cytochrome of R. rubrum was solubilized by a Triton X-100 treatment of chromatophores and was partly purified. It was found to have a molecular weight of about 17,000, a midpoint oxidation-reduction potential of -192 mV, and an alpha-absorption peak at 552 nm. It appears that low-potential c-type cytochromes may be present in all purple photosynthetic bacteria, of both the sulfur and the nonsulfur types.
Physiological role and membrane lipid modulation of the membrane-bound (Mg2+, na+)-adenosine triphosphatase activity in Acholeplasma laidlawii
Authors: Jinks D C , Silvius J R , McElhaney R N
Published: January 2020

The membrane-bound adenosine triphosphatase (ATPase) activity of Acholeplasma laidlawii B differs in many respects from the common (Mg2+, Ca2+)-ATPase activity of higher bacteria, most notably in that it is specifically activated by Mg2+ and strongly and specifically stimulated by Na+ (or Li+). Various inhibitors diminish the ATPase activity with a concentration dependence which suggests that a single enzyme species is responsible for all of the observed ATP hydrolytic activity (both basal and Na+ stimulated). The Km for ATP is influenced by temperature but not by membrane lipid fatty acid composition. Vmax is influenced by both of these factors, showing a break in Arrhenius plots which falls below the lipid phase transition midpoint but well above the lower boundary when a phase transition occurs within the temperature range studied. The apparent energy of activation for Vmax is strongly influenced by lipid fatty acid composition both above and below the break. When whole cells of A. laidlawii B are incubated in KCl or NaCl buffers, they rapidly swell and lyse if deprived of an energy source or treated with ATPase inhibitors at concentrations which significantly inhibit enzyme activity in isolated membranes, whereas in sucrose or MgSO4 buffers of equal osmolarity, the cells are stable under these conditions. These results suggest that the membrane ATPase of A. laidlawii B is intimately associated with the membrane lipids and that it functions as a monovalent cation pump which regulates intracellular osmolarity as the (Na+, K+)-ATPase does in eucaryotes.
Properties of lambda transducing bacteriophages carrying R100 plasmid DNA: mercury resistance genes
Authors: Dempsey W B , McIntire S A , Willetts N , Schottel J , Kinscherf T G , Silver S , Shannon W A
Published: January 2020

Three lambdamer (resistance to Hg2+ and mercurials) transducing phages were prepared from three independent cointegrate isolates of bacteriophage lambda and plasmid R100. DNA heteroduplex and restriction nuclease analyses of the lambdamer DNA showed that all three phages had resulted from lambda insertion at kilobase coordinate 8.6 of plasmid R100, followed by loss of different lengths of lambda DNA and replacement with different lengths of R100 DNA. Two of the lambdamer phages were defective, containing deletions from lambdaatt through the lambdaN gene and into the lambdarex gene; the third, VAlambda14, was an N+ Spi- plaque-forming phage. With VAlambda14, N-dependent transcription of R100 mer from the lambdapL promoter suggested that transcription of mer proceeded in the direction from IS1b toward the sulfonamide resistance determinant (i.e., from a plasmid promoter in restriction nuclease fragment EcoRI-H toward fragment EcoRI-I). Phage-directed protein synthesis in a UV-irradiated lambdaind- lysogen showed the Hg2+-inducible synthesis of three major polypeptides of molecular weights 68,000, 11,500, and 8,500 and three minor ones of molecular weights 54,000, 33,000, and 13,500. The largest of the major polypeptides is identified as the subunit of the mercuric reductase enzyme. The functions of the smaller polypeptides are not known. Hg2+ reductase enzyme assays confirmed the regulation of mer synthesis during phage infection.
malB region in Escherichia coli K-12: specialized transducing bacteriophages and first restriction map
Authors: Marchal C , Greenblatt J , Hofnung M
Published: January 2020

By starting from an Escherichia coli K-12 strain with a lambda phage integrated in the malB region, series of transducing phages carrying part or all of the malB region have been isolated. Genetic mapping of the transduced malB fragments was accomplished by complementation and recombination with known mutations in the region. By using the DNA of these phages, it was found that the malB region is cleaved by the restriction enzymes BglII, EcoRI, HaeII, HincII, SalI, and SstI, but not BamHI, HindIII, KpnI, PstI, XbaI, or XhoI. A physical map was constructed and tentatively correlated with the genetic map.
Meso-alpha,epsilon-diaminopimelate D-dehydrogenase: distribution and the reaction product
Authors: Misono H , Togawa H , Yamamoto T , Soda K
Published: January 2020

A high activity of meso-alpha-epsilon-diaminopimelate dehydrogenase was found in extracts of Bacillus sphaericus, Brevibacterium sp., Corynebacterium glutamicum, and Proteus vulgaris among bacteria tested. B. sphaericus IFO 3525, in which the enzyme is most abundant, was chosen to study the enzyme reaction. The enzyme was not induced by the addition of meso-alpha-epsilon-diaminopimelate to the growth medium. The reaction product was isolated and identified as alpha-amino-epsilon-ketopimelate by a comparison of the properties of its 2,4-dinitrophenylhydrazone with those of an authentic sample in silica gel thin-layer chromatography, absorption, infrared and proton nuclear magnetic resonance spectrometry, and elemental analyses. The alpha-amino-epsilon-ketopimelate formed enzymatically was decarboxylated by H2O2 to yield L-alpha-aminoadipate. This suggests that the amino group with D-configuration in the substrate is oxidatively deaminated; the enzyme is a D-amino acid dehydrogenase. L-alpha-Amino-epsilon-ketopimelate undergoes spontaneous dehydration to the cyclic delta1-piperideine-2,6-dicarboxylate. The enzyme reaction is reversible, and meso-alpha-epsilon-diaminopimelate was formed in the reductive amination of L-alpha-epsilon-ketopimelate.
Rifampin-resistant mutants of Myxococcus xanthus defective in development
Authors: Rudd K , Zusman D R
Published: January 2020

Rifampin, an antibiotic which is known to bind to and inhibit RNA polymerase, was used to probe the molecular regulation of development in Myxococcus xanthus. Rifampin-resistant mutants were screened for defects in fruiting-body formation. About 20% of the isolates in the initial screenings showed major defects in developmental aggregation or sporulation. Eleven independent mutants with wild-type growth rates and stable phenotypes were analyzed by transduction. In these strains, the rifampin-resistant and nonfruiting phenotypes showed cotransduction frequencies equal to or greater than 99.0 to 99.9%. The RNA polymerase activities were resistant to rifampin in vitro, indicating that the RNA polymerase is altered in these strains. Although their fruiting phenotypes are heterogeneous, these strains can be divided into two classes based on the level of aggregation. The results suggest that RNA polymerase plays a significant role in the regulation of development in M. xanthus since mutations which cause no apparent changes in vegetative growth result in striking defects in fruiting-body formation.
Incorporation of phosphatidylglycerol into murein lipoprotein in intact cells of Salmonella typhimurium by phospholipid vesicle fusion
Authors: Chattopadhyay P K , Lai J S , Wu H C
Published: January 2020

The biosynthesis of the diglyceride moiety of murein lipoprotein was studied by fusion of labeled phospholipid vesicles with intact cells of Salmonella typhimurium. Phosphatidylglycerol was found to be an excellent donor for the glyceryl moiety in lipoprotein, whereas phosphatidylethanolamine and cardiolipin were not. The incorporation of radioactivity from monoacyl-phosphatidylglycerol into lipoprotein can be attributed to its conversion to phosphatidylglycerol. The results strongly support our hypothesis that the glyceryl residue covalently linked to murein lipoprotein is derived from the nonacylated glycerol moiety of phosphatidylglycerol.
Isolation of an iron-binding compound from Pseudomonas aeruginosa
Authors: Cox C D , Graham R
Published: January 2020

An iron-binding compound was isolated from ethyl acetate extracts of culture supernatant fluids of Pseudomonas aeruginosa and was purified by successive paper and thin-layer chromatographic procedures. The purified compound was characterized by UV, visible, infrared, and fluorescence spectroscopy. The compound possesses phenolic characteristics, with little or no similarity to dihydroxybenzoates and no indication of a hydroxamate group. P. aeruginosa synthesized the compound during active growth in culture media containing less than 5 X 10(-6) M added FeCl3. When added to iron-poor cultures of P. aeruginosa, the compound promoted the growth of the bacterium and also reversed growth inhibition by the iron chelator ethylenediamine-di-(o-hydroxyphenylacetic acid).
Deletion analysis of the expression of rRNA genes and associated tRNA genes carried by a lambda transducing bacteriophage
Authors: Morgan E A , Nomura M
Published: January 2020

Transducing phage lambdailv5 carries genes for rRNA's, spacer tRNA's (tRNA1 Ile and tRNA1B Ala), and two other tRNA's (TRNA1 Asp and tRNA Trp). We have isolated a mutant of lambdailv5, lambdailv5su7, which carries an amber suppressor mutation in the tRNA Trp gene. A series of deletion mutants were isolated from the lambdailv5su7 phage. Genetic and biochemical analyses of these deletion mutants have confirmed our previous conclusion (E. A. Morgan, T. Ikemura, L. Lindahl, A. M. Fallon, and M. Nomura, Cell 13:335--344, 1978) that the genes for tRNA1 Asp and tRNA Trp located at the distal end of the rRNA operon (rrnC) are cotranscribed with other rRNA genes in that operon. In addition, these deletions were used to define roughly the physical location of the promoter(s) of the rRNA operon carried by the lambdailv5su7 transducing phage.
Vibrio cholerae hybrid sex factor that contains ampicillin transposon Tn1
Authors: Johnson S R , Romig W R
Published: January 2020

The ampicillin resistance transposon Tn1 was translocated from the R plasmid RP4 to the Vibrio cholerae conjugative plasmid, P. The hybrid sex factor P::Tn1 was highly transmissible and expressed the biological activities of the P factor. In addition, P::Tn1 facilitated transfer of RP4 to V. cholerae recipients. Physical studies of P::Tn1 indicated that the Tn1 transposon was added to the otherwise unaltered P plasmid.
Bacteriophage T4D receptors and the Escherichia coli cell wall structure: role of spherical particles and protein b of the cell wall in bacteriophage infection
Authors: Zorzopulos J , Kozloff L M , Chapman V , DeLong S
Published: January 2020

The nature of the interaction of bacteriophage T4D and the outer cell wall of its host, Escherichia coli B, has been investigated. Bacteria with altered or modified cell walls have been obtained by two different growth procedures: (i) growth in high osmolarity medium or (ii) growth in broth in the presence of divalent heavy metal ions. When these altered host cells were washed and subsequently added to regular growth medium, they interacted with added phage particles, but successful infection did not occur. Most of the phage particles released from these treated cells were observed to have full heads and an altered tail structure. The altered phage tails had contracted sheaths and unusual pieces of the bacterial cell wall attached to the distal portion of the exposed phage tail tube. Phage released from bacteria grown in the high osmolarity medium had attached cell wall pieces of two major types, these pieces being either 40 or 21 nm in diameter. The smaller-type cell wall pieces (21 nm) were formed by three spheres each measuring 7 nm in diameter. Phage particles released from cells previously exposed to the divalent metal ions had only one 7-nm cell wall sphere attached to the distal end of the tail tube. It was found that these 7-nm spheres (i) are normal components of the cell wall and are morphologically similar to endotoxin, (ii) are held in place on the cell wall by a component of the cell wall called protein b, and (iii) are most likely the site of penetration of the phage tail tube through which the phage DNA enters the host cell.
Direct Comparison of the Subtilisin-Like Intracellular Protease of
Bacillus licheniformis
with the Homologous Enzymes of
Bacillus subtilis
Authors: Strongin A. Ya. , Abramov Z. T. , Yaroslavtseva N. G. , Baratova L. A. , Shaginyan K. A. , Belyanova L. P. , Stepanov V. M.
Published: January 2020

Intracellular serine proteases of
Bacillus licheniformis
and
Bacillus subtilis
are closely related.
Increased levels of dihydrofolate reductase in rifampin-resistant mutants of Bacillus subtilis
Authors: Kane J F , Wainscott V J , Hurt M A
Published: January 2020

Several independent, spontaneous rifampin-resistant mutants of Bacillus subtilis were isolated and found to have an increased resistance to trimethoprim, an inhibitor of dihydrofolate reductase. This increased resistance in the rif mutants was the result of a specific threefold increase in the activity of dihydrofolate reductase, since six other enzymes examined remained unchanged. This increased level of dihydrofolate reductase and the trimethoprim resistance were cotransformed (100%) with the rif marker. These results suggest that the RNA polymerase is altered in its recognition of the gene that specifies dihydrofolate reductase.
A plasmid in Streptococcus pneumoniae
Authors: Smith M D , Guild W R
Published: January 2020

Plasmid deoxyribonucleic acid has been detected in three related laboratory strains of Streptococcus pneumoniae. Strains D39S, R36, and R36NC each contain a minimum of two copies per cell of a 2.0-megadalton plasmid (pDP1). A plasmid twice as large as this smaller one is also present in much lower quantity in these strains, but neither plasmid is present in four strains related to these or in a drug-resistant clinical isolate from Paris. The plasmid yield was not amplified in the presence of chloramphenicol. No phenotype has been correlated with the presence of pDP1, which has existed in strains carried for many years in laboratory collections.
Purification and Properties of Acetate Kinase from
Acholeplasma laidlawii
Authors: Kahane Itzhak , Muhlrad Andras
Published: January 2020

Acetate kinase (EC 2.7.2.1) was purified from
Acholeplasma laidlawii
cytoplasm by a combination of ammonium sulfate fractionation, gel filtration, diethylaminoethyl-cellulose chromatography, and affinity chromatography on 8-(6-aminohexylamino)-adenosine 5′-triphosphate conjugated to Sepharose 4B. The enzyme was composed of polypeptide chains of about 50,000 molecular weight as estimated from sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Under nondenaturating conditions, apparent molecular weights between 64,000 and 130,000 were obtained, depending upon mainly the ionic strength of the test solution. The enzyme had a narrow specificity for phosphate acceptor acids, whereas both purine and pyrimidine nucleoside triphosphates were suitable phosphate donors. Na
+
and K
+
inhibited both acetyl phosphate and adenosine 5′-triphosphate synthesis, and the latter was also inhibited by high concentrations of adenosine 5′-diphosphate and acetyl phosphate. This substrate inhibition was partially abolished by 0.5 M NaCl. The enzyme catalyzed the independent adenosine 5′-diphosphate↔adenosine 5′-triphosphate and acetate↔acetyl phosphate exchanges. The rate of the latter was enhanced by the addition of cosubstrate Mg
2+
–adenosine 5′-triphosphate. The high affinity for substrates, except for acetate, indicated that under physiological conditions the direction of the enzymic reaction favors adenosine 5′-triphosphate synthesis. Thus, a mechanism for adenosine 5′-triphosphate generation in mycoplasmas is suggested.
Purification and properties of the manganese-dependent phosphoglycerate mutase of Bacillus subtilis
Authors: Watabe K , Freese E
Published: January 2020

Phosphoglycerate mutase of Bacillus subtilis was purified to apparent homogeneity. It specifically required manganese ions for stability and activity, but it does not need 2,3-diphosphoglycerate as cofactor; the Km for Mn2+ is about 4.5 micrometer. Enzyme activity was inhibited by heavy-metal ions, 2,3-butanedione, and sulfhydryl agents. The mutase has a molecular weight of about 74,000 as shown by Sephadex gel filtration and by acrylamide gel electrophoresis in the presence of sodium dodecyl sulfate; it consisted of one polypeptide.
Analysis of corynomycolic acids and other fatty acids produced by Corynebacterium lepus grown on kerosene
Authors: Cooper D G , Zajic J E , Gracey D E
Published: January 2020

The saponifiable carboxylic acids of the extracellular product of Corynebacterium lepus grown on kerosene have been isolated and characterized. About 25% of these acids were a mixture of simple, saturated fatty acids ranging from C13 to C24 and including both even and odd homologues. The distribution of these acids was bimodal, with maxima at C15 and C21. The other 75% of the acids was a mixture of corynomycolic acids [R1--CH(OH)--CH(R2)--COOH] ranging from C28 to C43. The R1 alkyl fragments varied from C16 to C25, and R2 fragments varied from C6 to C14. Both even and odd corynomycolic acid homologues were observed, and the distribution had a single pronounced maximum at C32 and C33. Bacterial utilization of the carboxylic oxidation products of the kerosene substrate is suggested to account for the wide distribution in chain length of these saturated fatty acids and for the observation of both even and odd homologues.
Diglyceride Kinase Mutants of
Escherichia coli
: Inner Membrane Association of 1,2-Diglyceride and Its Relation to Synthesis of Membrane-Derived Oligosaccharides
Authors: Raetz Christian R. H. , Newman Karl F.
Published: January 2020

Mutants of
Escherichia coli
defective in diglyceride kinase contain 10 to 20 times more
sn
-1,2-diglyceride than normal cells. This material constitutes about 8% of the total lipid in such strains. We now report that this excess diglyceride is recovered in the particulate fraction, primarily in association with the inner, cytoplasmic membrane. The diglyceride kinase of wild-type cells was recovered in the same inner membrane fractions. The conditions employed for the preparation of the membranes did not appear to cause significant redistribution of lipids and proteins. The biochemical reactions leading to the formation of diglyceride in
E. coli
are not known. To determine whether diglyceride formation requires concurrent synthesis of the membrane-derived oligosaccharides (H. Schulman and E. P. Kennedy, J. Biol. Chem.
252:
4250-4255, 1977), we have constructed a double mutant defective in both the kinase (
dgk
) and phosphoglucose isomerase (
pgi
). When oligosaccharide synthesis was inhibited in this organism by growing the cells on amino acids as the sole carbon source, the diglyceride was no longer present in large amounts. When glucose was also added to the medium, the
pgi
mutation was bypassed, oligosaccharide synthesis resumed, and diglyceride again accumulated. These findings suggest that diglyceride may arise during the transfer of the
sn
-glycero-1-P moiety from phosphatidylglycerol (and possibly cardiolipin) to the oligosaccharides. In wild-type cells the kinase permits the cyclical reutilization of diglyceride molecules for phospholipid biosynthesis.
Multiple Intracellular Peptidases in
Neurospora crassa
Authors: Tan Sai-Tee , Marzluf George A.
Published: January 2020

Neurospora crassa
possesses multiple intracellular peptidases which display overlapping substrate specificities. They were readily detected by an in situ staining procedure for peptidases separated in polyacrylamide gels, within which the auxilliary enzyme,
l
-amino acid oxidase, was immobilized. Eleven different intracellular peptidases were identified by electrophoretic separation and verified by their individual patterns of substrate specificities. Most peptide substrates tested were hydrolyzed by several different peptidases. The multiple intracellular peptidases may play overlapping roles in several basic cell processes which involve peptidase activity. The amount of peptidase activity for leucylglycine present in crude extracts of cells grown under widely different conditions was relatively constant, suggesting that this enzyme may be constitutive, although alterations in the amounts of individual peptidase isozymes may occur. A single enzyme, designated peptidase II, was partially purified and obtained free from the other peptidase species. Peptidase II was found to be an aminopeptidase with activity toward many peptides of varied composition and size. It was more active with tripeptides than homologous dipeptides and showed strong activity toward methionine-containing peptides. This enzyme, with a molecular weight of about 37,000, was thermolabile at 65°C and was strongly inhibited by
p
-hydroxymercuribenzoate, Zn
2+
, Co
2+
, and Mn
2+
, but was insensitive to the serine protease inhibitor phenylmethylsulfonyl fluoride. Peptidase II apparently possesses an essential sulfhydryl group and may be a metalloenzyme.
Immunoelectron microscopic localization of penicillinase in Bacillus licheniformis
Authors: Ghosh A , Ghosh B K
Published: January 2020

Penicillinase was localized in log-phase cells of Bacillus licheniformis 749/C by labeling with ferritin-anti-penicillinase immunoglobulin G conjugate. Mildly fixed homogenized cells, isolated subcellular fractions, and frozen thin sections were labeled. The label was distributed in discrete patches in the cell envelope. The patches extended from the inside part of the membrane to the outside part of the wall. The inside part of the membrane was labeled more extensively than the outside part. The cytoplasm also bound some ferritin-immunoglobulin G conjugate. Immunoelectrophoresis and biochemical assay of cytosol material suggest that the cytoplasmic antigenic sites are a protease-sensitive form of penicillinase.
Nucleoside Diphosphate Sugar Hydrolase Gene of
Salmonella typhimurium
: Chromosomal Location Determined by Intergeneric Crosses
Authors: Minton N. , Gunn J. , Beacham I. R.
Published: January 2020

The gene specifying a membrane-bound nucleoside diphosphate sugar hydrolase of
Salmonella typhimurium
was mapped near the
metA
locus by using intergeneric crosses between this bacterium and
Escherichia coli
.
Change from Homo- to Heterolactic Fermentation by
Streptococcus lactis
Resulting from Glucose Limitation in Anaerobic Chemostat Cultures
Authors: Thomas Terence D. , Ellwood Derek C. , Longyear V. Michael C.
Published: January 2020

Lactic streptococci, classically regarded as homolactic fermenters of glucose and lactose, became heterolactic when grown with limiting carbohydrate concentrations in a chemostat. At high dilution rates (
D
) with excess glucose present, about 95% of the fermented sugar was converted to
l
-lactate. However, as
D
was lowered and glucose became limiting, five of the six strains tested changed to a heterolactic fermentation such that at
D
= 0.1 h
−1
as little as 1% of the glucose was converted to
l
-lactate. The products formed after this phenotypic change in fermentation pattern were formate, acetate, and ethanol. The level of lactate dehydrogenase, which is dependent upon ketohexose diphosphate for activity, decreased as fermentation became heterolactic with
Streptococcus lactis
ML
3
. Transfer of heterolactic cells from the chemostat to buffer containing glucose resulted in the nongrowing cells converting nearly 80% of the glucose to
l
-lactate, indicating that fine control of enzyme activity is an important factor in the fermentation change. These nongrowing cells metabolizing glucose had elevated (ca. twofold) intracellular fructose 1,6-diphosphate concentrations ([FDP]
in
) compared with those in the glucose-limited heterolactic cells in the chemostat. [FDP]
in
was monitored during the change in fermentation pattern observed in the chemostat when glucose became limiting. Cells converting 95 and 1% of the glucose to
l
-lactate contained 25 and 10 mM [FDP]
in
, respectively. It is suggested that factors involved in the change to heterolactic fermentation include both [FDP]
in
and the level of lactate dehydrogenase.
Combined use of strain construction and affinity chromatography in the rapid, high-yield purification of 6-phosphogluconate dehydrogenase from Escherichia coli
Authors: Wolf R E , Shea F M
Published: January 2020

A rapid, high-yield method for purification of 6-phosphogluconate dehydrogenase from Escherichia coli K-12 is described. Sonic extracts prepared from heat-induced cultures of strain RW184, doubly lysogenic for the specialized transducing bacteriophage lambdacI857St68h80dgndhis and bearing a deletion of the gene for glucose 6-phosphate dehydrogenase, contained levels of 6-phosphogluconate dehydrogenase 15- to 20-fold higher than cultures of wild-type cells. Affinity chromatography on blue dextran-Sepharose with batchwise elution with 1 mM nicotinamide adenine dinucleotide phosphate affected a further 10-fold purification. Enzyme prepared in this manner was homogeneous according to electrophoresis on sodium dodecyl sulfate-polyacrylamide gels and immunoelectrophoresis using antiserum directed against it. Fructose 1,6-diphosphate is an inhibitor of enzyme activity.
Chemical alterations in cell envelopes of polymyxin-resistant Pseudomonas aeruginosa isolates
Authors: Gilleland H E , Lyle R D
Published: January 2020

Cell envelopes from Pseudomonas aeruginosa strains resistant to polymyxin were compared with cell envelopes from polymyxin-sensitive strains as to their content of total protein, carbohydrate, and 2-keto-3-deoxyoctonate and as to their protein composition as determined by slab polyacrylamide gel electrophoresis. The cell envelopes of the polymyxin-resistant strains had reduced amounts of lipopolysaccharide, as indicated a reduction in both carbohydrate and 2-keto-3-deoxyoctonate concentrations, and a greatly altered protein composition as shown by polyacrylamide gel electrophoresis. There was a quantitative increase in total cell envelop protein in these strains. However, those protein bands identified as being major outer membrane proteins upon polyacrylamide gel electrophoresis of separated outer and cytoplasmic membranes were reduced greatly in concentration in the polymyxin-resistant cell envelopes. Thus, it appears that polymyxin resistance in these strains is associated with the alteration of the outer membrane through a loss of lipopolysaccharide and outer membrane proteins.
Genetic recombination during transformation in Bacillus subtilis: appearance of a deoxyribonucleic acid methylase
Authors: Ganesan A T
Published: January 2020

In Bacillus subtilis the ability to take up deoxyribonucleic acid (DNA) and undergo genetic transformation may coincide with the induction of defective phage(s) and the expression of possibly related cryptic genes. A restriction-modification enzyme system appears to be expressed. Targets of the restriction activity on the DNA can be blocked my methylation catalyzed by the methyl transferase. It is shown that cellular DNA becomes progressively methylated and reaches the maxium level during the peak of competency. Deoxycytidine residues of both incoming donor and resident DNA are methylated. The possible participation of these enzymes in recombination and the general role of cryptic genes in inducible functions are discussed.
Amylooligosaccharides from Mycobacterium smegmatis
Authors: Yamada H , Ballou C E
Published: January 2020

In early stationary phase of growth, Mycobacterium smegmatis cultures accumulate amylooligosaccharides (alpha 1 leads to 4-glucooligosaccharides) up to the undecasaccharide. Although M. smegmatis also makes an acylated polymethylpolysaccharide that is predominantly and alpha 1 leads to 4-glucan, we conclude that these oligosaccharides are precursors of glycogen rather than lopopolysaccharide.
Uptake and catabolism of D-xylose in Salmonella typhimurium LT2
Authors: Shamanna D K , Sanderson K E
Published: January 2020

Salmonella typhimurium LT2 grows on D-xylose as sole carbon source with a generation time of 105 to 110 min. The following activities are induced at the indicated time after the addition of the inducer, D-xylose: D-xylulokinase (5 min), D-xylose isomerase (7 to 8 min), and D-xylose transport (10 min). All other pentoses and pentitols tested failed to induce isomerase or kinase. Synthesis of D-xylose isomerase was subject to catabolite repression, which was reversed by the addition of cyclic adenosine monophosphate. Most of the radioactive counts from D-[14C]xylose were initially accumulated in the cell in the form of D-xylose or D-xylulose. D-Xylose uptake in a mutant which was deficient in D-xylose isomerase was equal to that of the wild type. The apparent Km for D-xylose uptake was 0.41 mM. Some L-arabinose was accumulated in D-xylose-induced cells, and some D-xylose was accumulated in L-arabinose-induced cells. D-Xylitol and L-arabinose competed against C-xylose uptake, but D-arabinose, D-lyxose, and L-lyxose did not. Osmotic shock reduced the uptake of D-xylose by about 50%; by equilibrium dialysis, a D-xylose-binding protein was detected in the supernatant fluid after spheroplasts were formed from D-xylose-induced cells.
Genetics and regulation of D-xylose utilization in Salmonella typhimurium LT2
Authors: Shamanna D K , Sanderson K E
Published: January 2020

Twenty-one Xyl- mutants of Salmonella typhimurium were selected: all had lost one or more of the activities for D-xylose isomerase, C-xylulokinase, or D-xylose transport. The mutants were classified into five functional groups: xylR, pleiotropic negative (12 mutants); xylA, D-xylose isomerase defective (3 mutants); xylB, D-xylulokinase defective (2 mutants); xylT, D-xylose transport defective (1 mutant); and 3 mutants with defective D-xylose isomerase and D-xylulokinase. Some nonsense mutations were identified among the xylR mutants. Two F'xyl plasmids were isolated by selection for early transfer of xyl+ by an Hfr which transfers xyl as a terminal gene; a plasmid with a mutation in the xyl genes, F'xylR1, was also isolated. Complementation tests using F'xyl plasmids indicate that expression of the xylA, xylB, and xylT genes is under the positive control of the xylR regulatory gene. Conjugation crosses and P22-mediated transduction data indicate that all the xyl mutations tested are in a cluster of genes at 78 units on the linkage map, and that the gene order is xylT--xylR--xylB--xylA--glyS--mtlA,D.
Characteristics of Anabaena variabilis influencing plaque formation by cyanophage N-1
Authors: Currier T C , Wolk C P
Published: January 2020

Phage N-1 grown in Anabaena strain 7120 [N-1 . 7120] forms plaques on A. variabilis about 10(-7) to 10(-6) as efficiently as on Anabaena 7120. By manipulating different characteristics of the interaction between phage and host, it was possible to increase the relative efficiency of plaque formation to 0.38. Growth of A. variabilis at 40 degrees C for at least three generations resulted in an increase in the rate of phage adsorption and a 10-fold increase in the efficiency of plaque formation. The efficiency of plaque formation was further increased about 42-fold, with little or no further increase in rate of adsorption, in a variant strain. A. variabilis strain FD, isolated from a culture of A. variabilis which had grown for more than 30 generations at 40 degrees C. The low relative efficiency of plaque formation by N-1 . 7120 on A. variabilis could be partially accounted for if A. variabilis contains a deoxyribonucleic acid restriction endonuclease which is absent from Anabaena 7120. Indirect evidence for such an endonuclease included the following: (i) phage N-1 grown in A. variabilis (N-1 . Av) had approximately a 7 X 10(3)-fold higher relative efficiency of plaque formation on A. variabilis than had N-1 . 7120; and (ii) the efficiency of plaque formation by N-1 . 7120 on A. variabilis strain FD was increased by up to 146-fold after heating the latter organism at 51 degrees C.
Transposition of ampicillin resistance to an enterotoxin plasmid in an Escherichia coli strain of human origin
Authors: McConnell M M , Willshaw G A , Smith H R , Scotland S M , Rowe B
Published: January 2020

We examined a strain of Escherichia coli, serotype O159.H34, of human origin which produced heat-stable and heat-labile enterotoxins, was resistant to ampicillin, and produced colicin. By conjugation and transformation experiments plasmids coding for enterotoxin production (Ent), enterotoxin production and ampicillin resistance (Ap-Ent), ampicillin resistance (Ap), and colicin production were isolated. Both the Ent and Ap-Ent plasmids were autotransferring and belonged to the F-incompatibility complex. However, the Apr Ent+ transconjugants showed differences in their levels of resistance and in their abilities to propagate F-specific phages and to transfer resistance. The results suggested there was transposition from the small Ap plasmid to the Ent plasmid. The Ap-Ent plasmids were larger than the enterotoxin factor and when treated with restriction endonuclease BamHI showed an additional fragment not present in the enterotoxin plasmid. The insertion of ampicillin resistance probably occurred at different sites on the enterotoxin plasmid, resulting in the observed variation in phenotype.
Escherichia coli mutant strain with altered expression of the tryptophan operon: ribonucleic acid synthesis in vitro
Authors: Pouwels P H , de Groot G P
Published: January 2020

Ribonucleic acid (RNA) synthesis has been studied in vitro with a partially purified preparation of RNA polymerase from a mutant strain of Escherichia coli with a reduced rate of accumulation of tryptophan RNA (P.H. Pouwels and H.J. Scholten, J. Bacteriol. 139:393-397, 1979). The incorporation of radioactive label into RNA with polymerase from mutant bacteria is considerably lower than that with the enzyme from wild-type bacteria. These results are explained by the presence in mutant bacteria, but not in wild-type bacteria, of a factor which suppresses the accumulation of RNA. Mutant bacteria contain a factor which renders RNA synthesis with mutant and wild-type RNA polymerase resistant to various inhibitors of RNA synthesis, e.g. rifampin, streptolydigin, and heparin. We conclude that in mutant bacteria a factor is modified which suppresses the accumulation of some RNA species and lowers the sensitivity of RNA polymerase to some transcription inhibitors.