bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic and physiological analysis of an envB spherelike mutant of Escherichia coli K-12 and characterization of its transductants
Authors: Westling-Häggström B , Normark S
Published: January 2020

The envB1 mutation mediating a distorted cell morphology of Escherichia coliK-12 was cotransducible with strA, aroE, aspB, and argG. The mapping data is consistent with a gene location for envB around 62.5 min. In partial diploids envB1 was recessive to its wild-type allele. The original envB mutant contained a second mutation in a locus denoted sloB close to strA. The following gene order is suggested: sloB-strA-aroE-envB-aspB-argG. The sloB1 mutation caused a marked reduction in the growth rate of both envB and envB+ strains. Moreover, this mutation in the presence of envB1 appears to increase the ratio between deoxyribonucleic acid and protein in cells growing in rich medium. The phenotypic properties of envB1, sloB+ and envB+ transductants were characterized. Cells with envB1, sloB+ genotype were hypersensitive to several penicillins including the beta-lactam compound, amidino penicillin. Penicillin hypersensitivity could not be explained by increased outer membrane penetrability. The original envB mutant (envB1,SLOB1), as well as envB1, sloB1 or envB+, SLOB1 transductants were resistant to amidino penicillin. Resistance was explained by the slow growth rate mediated by the sloB1 mutation. The similarity between envB cells and wild-type cells treated with sublethal concentrations of amidino penicillin was emphasized.
Regulation of nitrogen metabolism in Escherichia coli and Klebsiella aerogenes: studies with the continuous-culture technique
Authors: Senior P J
Published: January 2020

Ammonia-nitrogen-limited continuous cultures of Escherichia coli and Klebsiella aerogenes contain induced levels of glutamine synthetase that is deadenylyated (i.e., fully active). In the presence of excess ammonia or glutamate in glucose-limited cultures of E. coli, glutamine synthetase is repressed and adenylylated (inactive). The average state of adenylylation (n) is a linear function of the specific growth rate. At low specific growth rates, glutamine synthetase is adenylylated; as the specific growth rate increases, n decreases, approaching 0 to 2 at rapid growth rates. The average state of adenylylation correlates well with the intracellular concentrations and ratios of alpha-ketoglutarate and glutamine, which are key effectors in the adenylylation-deadenylylation systems. E. coli and K. aerogenes differ markedly in their growth yields, growth rates, and enzymatic composition during nitrogen limitation. The data suggest that, unlike K. aerogenes, E. coli W uses glutamate dehydrogenase to incorporate ammonia during nitrogen limitation. In E. coli, glutamate dehydrogenase is progressively induced during nitrogen limitation when mu (growth rate) approaches mumax. In contrast, in K. aerogenes glutamate dehydrogenase is repressed during nitrogen limitation, whereas glutamate synthase, an alternative supplier of glutamate to the cell, is induced. Data are presented that support the regulatory schemes proposed for the control of glutamine synthetase activity by induction-repression phenomena and adenylylation-deadenylylation reaction. We propose that the intracellular ratio of alpha-ketoglutarate to glutamine may be the most important physiological parameter in determining the activity of glutamine synthetase.
Passage of a membrane protein through the walls of toluene-treated Bacillus megaterium cells
Authors: Fan D P , Gardner-Eckstrom G L
Published: January 2020

Based on autoradiographic and microscopic evidence, it seems likely that a membrane protein essential for peptidoglycan synthesis can be extracted from uhlysed toluene-treated Bacillus megaterium cells. Furthermore, this protein can be added back to the membrane through the wall to reconstitute peptidoglycan synthesis. Autoradiograms also show that peptidoglycan is synthesized from externally added nucleotide precursors over the entire length of the toluene-treated bacterial. The amounts of peptidoglycan made is to small to be visible by thin section electron microscopy.
Regulation of glutaminase levels in Escherichia coli
Authors: Prusiner S
Published: January 2020

Nitrogenous metabolites, cyclic adenosine 3':5'-monophosphate (cAMP), and the stage of culture growth all influence the levels of glutaminase A in Escherichia coli, but no variables in culture conditions alter the levels of glutaminase B. Growth of E. coli on culture media containing glucose and excess ammonia results in a rise in the level of glutaminase A as the cultures enter stationary phase; this rise is abolished by ammonia limitation. cAMP or glycerol reduce the level of glutaminase A. In mutants deficient in cAMP receptor protein, glutaminase A levels are unchanged by cAMP, but they are still susceptible to regulation by ammonia. We consider glutaminase B to be a constitutive enzyme, since its levels appear independent of nutritional conditions.
Maltose transport in Escherichia coli K-12: involvement of the bacteriophage lambda receptor
Authors: Szmelcman S , Hofnung M
Published: January 2020

Mutants affected in lamB, the structural gene for phage lambda receptor, are unable to utilize maltose when it is present at low concentrations (less than or equal 10 muM). During growth in a chemostat at limiting maltose concentrations, the lamB mutants tested were selected against in the presence of the wild-type strain. Transport studies demonstrate that most lamB mutants have deficient maltose transport capacities at low maltose concentrations. When antibodies against purified phage lambda receptor are added to a wild-type strain, transport of maltose at low concentrations is significantly reduced. These results strongly suggest that the phage lambda receptor molecule is involved in maltose transport.
Role of methionine in the regulation of serine hydroxymethyltransferase in Eschericia coli
Authors: Greene R C , Radovich C
Published: January 2020

Significant derepression of serine hydroxymethyltransferase is observed when metE or metF mutants of Escherichia coli K-12 are grown on D-methionine sulfoxide instead of L-methionine. The derepression is not prevented by addition of glycine, adenosine, guanosine, guanosine, and thymidine to the growth medium of methionine-limited metF cells showing that the effect is not due to a secondary deficiency of these nutrients. On the other hand, methionine-limited growth of a metA mutant leads to derepression of met regulon enzymes, but only a marginal increase in serine hydroxymethyltransferase activity. A prototrophic metJ strain grown on minimal medium has about the same serine hydroxymethyltransferase as the wild type. The enzyme activity of the metJ strain is not influenced by methionine, but it is partially repressed by glycine, adenosine, and thymidine. metK strains have about twice as much serine hydroxymethyltransferase activity as wild-type cells when grown on minimal medium; but when both types of cells are grown on medium supplemented with glycine, adenosine, guanosine, and thymidine, their enzyme activities are about the same. The results show that methionine limitation can lead to depression of serine hydroxymethyltransferase, but that the regulatory system is different from the one which controls the methionine regulon.
Effects of temperature and nutritional changes on the fatty acids of agmenellum quadruplicatum
Authors: Olson G J , Ingram L O
Published: January 2020

The fatty acid composition of the blue-green bacterium Agmenellum quadruplicatum was examined under a wide variety of growth conditions. The fatty acid composition was found to undergo significant changes with variations in temperature, media composition, and growth phase (log versus stationary). With increasing growth temperature (20 to 43 C) log-phase cells exhibited an increase in saturated fatty acids (38.4% at 20 C to 63.6% at 43 C). Striking changes were seen with some of the individual fatty acids such as 18.3, which made up 16.0% of the total fatty acid at 20 C but was not neasurable at 43 C. Fatty acid 12:0 was not measurable at 20 C but made up 16.3% of the total fatty acids at 43 C. Cell lipids were separated into neutral lipid, glycolipid, and very polar liquid fractions. The neutral lipid fraction was composed almost entirely of 12 carbon fatty acids (12:0, 12:1). Glycolipid and very polar lipids were more similar in their fatty acid composition when compared to the total cellular fatty acids, although they did lack 12 carbon fatty acids. The total of 12 carbon fatty acids in the cell can be used as an indicator of the amount of neutral lipid present.
Metabolism of cyclic adenosine 3',5'-monophosphate and induction of tryptophanase in Escherichia coli
Authors: Botsford J L
Published: January 2020

The relationship between cyclic adenosine 3',5'-monophosphate (cyclic AMP) metabolism and the induction of tryptophanase and beta-galactosidase was studied in several strains of Escherichia coli grown with succinate, acetate, glycerol, or glucose as the carbon source. No consistent relationship between the intracellular concentration of cyclic AMP in the several strains cultured and the various carbon sources was discerned. In E. coli K-12-1 the induction of tryptophanase was found to vary in the order: succinate greater than acetate greater than glycerol greater than glucose, and that of beta-galactosidase was found in the order: glycerol greater than acetate greater than succinate greater than glucose. Rate of accumulation of cyclic AMP in the culture filtrate was in the order: succinate greater than acetate greater than glycerol greater than glucose. The addition of glycerol to E. coli K-12-1 grown in acetate caused inhibition of tryptophanase and slight inhibition of accumulation of extracellular cyclic AMP. These same conditions caused beta-galactosidase induction to be stimulated. The addition of exogenous cyclic AMP to cultures grown with four different carbon sources had an effect characteristic for each of the two enzymes studied as well as each individual carbon source. The results suggest that there are control elements distinct from cyclic AMP and its receptor protein which respond to the catabolic situation of the cell.
Membrane-mediated killing of Saccharomyces cerevisiae by glycoproteins from Torulopsis glabrata
Authors: Bussey H , Skipper N
Published: January 2020

Cell-free supernatants from cultures of Torulopsis glabrata contained glycoprotein toxins that killed sensitive and killer strains of Saccharomyces cerevisiae with single-hit kinetics. Growing S. cerevisiae treated with the toxins showed a leakage of cellular potassium, partial dissipation of the adenosine triphosphate pool, and a coordinate shutdown of macromolecular synthesis. These pool efflux-stimulating toxins have been partially purified and at least three toxic glycoproteins have been separated. Pool efflux-stimulating toxin activity was stable from pH 3 through 7, though killing was maximal close to pH 4.
Regulation of lactate dehydrogenase and change of fermentation products in streptococci
Authors: Yamada T , Carlsson J
Published: January 2020

Streptococcus mutans JC 2 produced mainly lactate as a fermentation product when grown in nitrogen-limited continuous culture in the presence of an excess of glucose and produced formate, acetate, and ethanol, but no lactate, under glucose-limited conditions. The levels of lactate dehydrogenase (LDH) in these cultures were of the same order of magnitude, and the activity of LDH was completely dependent on fructose-1,6-diphosphate (FDP). The intracellular level of FDP was high and the level of phosphoenolpyruvate (PEP) was low under the glucose-excess conditions. In the glucose-limited cultures, all glycolytic intermediates studied, except PEP, were low. S. mutans FIL, which had an FDP-independent LDH and similar levels of glycolytic intermediates as S. mutans JC2, produced mainly lactate under glucose-excess or under glucose-limited conditions. LDH of Streptococcus bovis ATCC 9809 was dependent on FDP for activity at a low concentration of pyruvate but had a significant activity without FDP at a high concentration of pyruvate. This strain also produced mainly lactate both under glucose-excess and glucose-limited conditions. The levels and characteristics of these LDHs were not changed by the culture conditions. These results indicate that changes in the intracellular level of FDP regulate LDH activity, which in turn influences the type of fermentation products produced by streptococci. PEP, adenosine 5'-monophosphate, adenosine 5'-diphosphate, and inorganic phosphate significantly inhibited LDH activity from S. mutans JC 2 and may also participate in the regulation of LDH activity in other streptococci.
CO
2
Production from Galactose in Galactose-1-Phosphate Uridyl Transferase-Deficient
Escherichia coli
Authors: LaPolla Robert J. , Geier Mark R. , Friedman Thomas B. , Merril Carl R.
Published: January 2020

Escherichia coli
K-12 deficient in galactose-1-phosphate uridyl transferase is capable of converting significant amounts of
d
-[1-
14
C]galactose to
14
CO
2
, whereas strains deficient in other enzymes of the Leloir pathway cannot do so.
Properties of the nicotinamide adenine dinucleotide phosphate-specific isocitrate dehydrogenase from Blastocladiella emersonii
Authors: Ingebretsen O C
Published: January 2020

The nicotinamide adenine dinucleotide phosphate (NADP)-specific isocitrate dehydrogenase from Blastocladiella emersonii was purified. The enzyme was very unstable. Satisfactory stability was obtained in the presence of 0.2% ovalbumin. The enzyme had a molecular weight of about 100,000. It did not exhibit homotropic cooperativity for any of it substrates and was not affected by the allosteric modifiers citrate and adenosine monophosphate, diphosphate, and tri-phosphate. The substrate saturation studies showed both intercept and slope effects in Lineweaver-Burk plots. The Km values for isocitrate and NADP were found to be 20 and 10 muM, respectively. The product inhibition pattern was compatible with a random sequential reaction mechanism. The enzyme catalyzed the oxidative decarboxylation of isocitrate about six times better than the reductive carboxylation of alpha-ketoglutarate. The enzyme was inhibited by glyoxylate plus oxalacetate. Assays conducted in the presence of low Mg2+ concentrations exhibited a lag. This lag could be abolished by the addition of reduced NADP to the assay mixture.
Metabolism of toluene and xylenes by Pseudomonas (putida (arvilla) mt-2: evidence for a new function of the TOL plasmid
Authors: Worsey M J , Williams P A
Published: January 2020

Pseudomonas putida (arvilla) mt-2 carries genes for the catabolism of toluene, m-xylene, and p-xylene on a transmissible plasmid, TOL. These compounds are degraded by oxidation of one of the methyl substituents via the corresponding alcohols and aldehydes to benzoate and m- and p-toluates, respectively, which are then further metabolised by the meta pathway, also coded for by the TOL plasmid. The specificities of the benzyl alcohol dehydrogenase and the benzaldehyde dehydrogenase for their three respective substrates are independent of the carbon source used for growth, suggesting that a single set of nonspecific enzymes is responsible for the dissimilation of the breakdown products of toluene and m- and p-xylene. Benzyl alcohol dehydrogenase and benzaldehyde dehydrogenase are coincidently and possible coordinately induced by toluene and the xylenes, and by the corresponding alcohols and aldehydes. They are not induced in cells grown on m-toluate but catechol 2,3-oxygenase can be induced by m-xylene.
Plasmid incompatibility and control of replication: copy mutants of the R-factor R1 in Escherichia coli K-12
Authors: Uhlin B E , Nordström K
Published: January 2020

Plasmid incompatibility was studied in Escherichia coli K-12. By double-antibiotic selection, clones were constructed that carried the two R-factors R1 and R100, both belonging to the compatibility group FII. After release of the selection pressure, each of the two plasmids was lost at the same rate (8% per generation). Mutants of R-factor R1 showing an increased number of copies per chromosome (copy mutants) were tested for their incompatibility towards R-factor R100. The results indicate that plasmid incompatibility is quantitative and not just a qualitative property. All copy mutants studied affected incompatibility, and there were two classes of mutants: one increasing and one decreasing the incompatiblity exerted towards the test plasmid R100. Evidence is presented that incompatibility is related to the mechanisms that control replication. The implications of such a relation on proposed models for control of replication are discussed. The data do not support the hypothesis that plasmid incompatibility is due to competition for a replicational or segregational site.
Tryptophan synthetase alpha(5.7-S): novel molecular species formed within Escherichia coli
Authors: Berger F G , Herrmann K M
Published: January 2020

A novel molecular species contributes about 5% of the total tryptophan synthetase of Escherichia coli derepressed for the trp operon enzymes. The new species is identified under conditions in which the dissociation of the two nonidentical subunits of the tryptophan synthetase complex is favored. The new species sediments at 5.7S, catalyzes the conversion of indole-3-glycerol phosphate to indole, and has been designated alpha(5.7-S). Although alpha(5.7-S) is not observed in extracts of trpA or trpB mutant strains deficient in the ability to form tryptophan synthetase alpha or beta2 subunits, respectively, a mixture of the two extracts allows the formation of alpha(5.7-S). Similar results are obtained when a homogeneous alpha protein is mixed with an extract of a trpA mutant strain, suggesting that the interaction of alpha and beta2 proteins is obligatory for alpha(5.7-S) formation. One can obtain a beta2 protein preparation that when mixed with a pure alpha protein gives no alpha(5.7-S). Therefore, the interaction of alpha and beta2 proteins alone is not sufficient for the formation of alpha(5.7-S). When a mixture of alpha and beta2 proteins devoid of alpha(5.7-S) is added to extracts of trp deletion mutants, the novel species can be reconstituted in vitro only when deletions are used that carry at least the operator-proximal part of the trpB gene. Therefore, it is concluded that the alpha(5.7-S) species of tryptophan synthetase results from the interaction of the alpha protein, the beta2 protein, and a third component, beta', specified by the deoxyribonucleic acid defined by the end points of two trp deletion mutants.
Formation and cleavage of 2-keto-3-deoxygluconate by 2-keto-3-deoxygluconate aldolase of Aspergillus niger
Authors: Allam A M , Hassan M M , Elzainy T A
Published: January 2020

2-Keto-3-deoxygluconate aldolase of Aspergillus niger, an enzyme that has not been reported previously, was purified 468-fold. Maximal activity was obtained at pH 8.0 and 50 C. The enzyme exhibited relative stereochemical specificity with respect to glyceraldehyde. The Km values for 2-keto-3-deoxygluconate, glyceraldehyde, and pyruvate were 10, 13.3, and 3.0 mM, respectively. The effects of some compounds and inhibitors on enzyme activity were examined. Stability of the enzyme under different conditions was investigated. The equilibrium constant was about 0.33 X 10(-3) M.
Operator-constitutive mutants in the threonine operon of Escherichia coli K-12
Authors: Saint-Girons I , Margarita D
Published: January 2020

Three Escherichia coli K-12 mutant strains resistant to DL-alpha-amino-beta-hydroxyvaleric acid were isolated in which the expression of the thr operon is constitutive. The localization and dominance properties of the mutations involved, called thrO, are those of operator mutations. The gene sequence is OABC as suggested by earlier studies.
Interference of dna ts mutations of Escherichia coli with thymineless death
Authors: Bouvier F , Sicard N
Published: January 2020

Thermosensitive mutants of Escherichia coli in which deoxyribonucleic acid replication is inhibited at high temperature have been assayed for their response to thymine deprivation at permissive and nonpermissive temperatures. The survival rate of mutants in which initiation of replication is defective at 42 C (dnaA and dnaC-D) is much higher at nonpermissive temperatures than at 32 C. By contrast, thymineless death is not hindered at 42 C in two elongation mutants (dnaE and dnaG). Bacterial strains belonging to the dnaB class exhibit both types of behavior. These results are in accordance with the model linking thymineless death to a disruption of an active replication fork. We discuss the possibility that the dnaB protein takes part in this process of cell decay.
Identification of poly-gamma-glutamyl chain lengths in folates of Bacillus subtilis
Authors: Hintze D N , Farmer J L
Published: January 2020

Bacillus subtilis strains 168 met ile leu and 23 thy contain folates which differ from one another in the number of glutamyl residues. The folate species were identified by reductive cleavage to the corresponding p-aminobenzoylglutamyl poly-gamma-glutamates and chromatography on diethylaminoethyl-cellulose. Pteroyltriglutamate is the predominant folate type, accounting for 86 to 88% of the total. Pteroyltetraglutamate is the only other type present in appreciable quantities, accounting for 5 to 6% of the total folates. Pteroyldiglutamate and pteroylpentaglutamate are present in small amounts, accounting for 1 to 3% and 1% of the total folates, respectively. Strain 168 met ile leu contains a very small amount of pteroylmonoglutamate (less than 0.5% of the total folates), but the other strain contains none.
Electron transport chain from glycerol 3-phosphate to nitrate in Escherichia coli
Authors: Miki K , Lin E C
Published: January 2020

It is known that in Escherichia coli two dehydrogenases of the flavoprotein kind can participate in the transfer of hydrogens from sn-glycerol 3-phosphate (G3P) to nitrate and that possession of either enzyme is sufficient to permit anaerobic growth on glycerol as carbon source and nitrate as hydrogen acceptor. Results from this study show that under such a growth condition a protein with light-absorption characteristics of cytochrome b1 is induced. If G3P, nitrate, and adenosine diphosphate are added anaerobically to a particulate fraction prepared from these cells, four reactions can be detected: (i) the reduction of the cytochrome b1-like protein, (ii) the formation of dihydroxyacetone phosphate (DHAP), (iii) the formation of nitrite, and (iv) the generation of adenosine 5'-triphosphate (ATP). The anaerobic G3P dehydrogenase system can yield an ATP-DHAP (or ATP-nitrite) molar ratio of about 0.2, whereas the aerobic G3P dehydrogenase system can yield a corresponding ratio of about 0.3. The hydrogen transfer activity is sensitive to respiratory inhibitors such as cyanide, Rotenone, and 2-heptyl-4-hydroxyquinoline-N-oxide.
Isolation and description of a menaquinone mutant from Bacillus licheniformis
Authors: Goodman S R , Marrs B L , Narconis R J , Olson R E
Published: January 2020

A menaquinone mutant (SG1) of Bacillus licheniformis has been isolated by selecting for colonies that are resistant to low levels of kanamycin (1.5 mug/ml) but sensitive to the same concentration of kanamycin in the presence of shikimate (25 mug/ml). The wild type (IU1) contained 0.38 +/- 0.02 nmol of menaquinone-7 (MK-7) per mg (dry weight) of cells when grown +/- shikimate, whereas SG1 had less than 0.01 nmol of MK-7 per mg (dry weight) of cells when grown in the presence of shikimate. SG1 had a generation time of 85 min, as compared to 24 min for IU1 grown +/- shikimate. SG1 doubled with a generation time of 28 min when grown in the presence of shikimate. IU1 consumed O2 at various rates depending on the stage of growth. A triphasic O2 consumption curve with maxima at mid-exponential phase, the transition from exponential to stationary phase, and early stationary phase was found for IU1 +/- shikimate and SG1 + shikimate. SG1 grown without shikimate consumed O2 at a low level (10 to 20% of IU1). Normal respiration could be restored to SG1 8.5 min after shikimate addition, whereas normal growth was not restored until 40 min after shikimate addition. Electron microscopic studies of SG1 and IU1 have indicated a morphological alteration in the mutant. SG1 is a dwarf cell as compared to IU1, when grown without shikimate. However, SG1 grown with shikimate became morphologically indistinguishable from IU1.
Direction of deoxyribonucleic acid replication in Escherichia coli under various conditions of cell growth
Authors: Rodriguez R L , Davern C I
Published: January 2020

The direction of chromosome replication in a temperature-sensitive initiation mutant of Escherichia coli (CT28) is shown autoradiographically to be bidirectional. This mode of replication persists even when the rate of replication is reduced by slow growth in succinate minimal medium or in the presence of chloramphenicol. Therefore, although the rate of replication can be affected by certain physiological stimuli, the topology of replication need not be.
Effect of
GAL
4 Gene Dosage on the Level of Galactose Catabolic Enzymes in
Saccharomyces cerevisiae
Authors: Klar Amar J. S. , Halvorson Harlyn O.
Published: January 2020

Lack of
GAL4
gene dosage on the level of uridine diphosphogalactose epimerase (EC 5.1.3.2) activity suggests the positive regulatory role for this locus on the control of galactose catabolic enzymes in
Saccharomyces cerevisiae
.
Properties of an inducible uptake system for beta-ketoadipate in Pseudomonas putida
Authors: Ornston L N , Parke D
Published: January 2020

Wild-type strains of Pseudomonas putida form an inducible uptake system that appears to act on beta-ketoadipate under normal physiological conditions. The system is induced by beta-ketoadipate and is represented by catabolites derived from it. Adipate is metabolized very slowly by wild-type P. putida cultures; [14C]adipate was used as an analogue of beta-ketoadipate to measure the transport activity in wild-type cells and in cells that constitutively produced the uptake system. Constitutive cells that contained high levels of the uptake system concentrated adipate to a level up to 200-fold above the concentration in the external medium. The process was energy dependent. The activity of the system with radioactive adipate was inhibited by beta-ketoadipate, by beta-ketoadipate analogues, and by some compounds (e.g., acetate, glucose) that are structurally unrelated to beta-ketoadipate; it is not known if the inhibitory effects are exerted directly by the compounds themselves or indirectly by catabolites derived from the compounds. The discovery of the beta-ketoadipate uptake system is surprising in view of earlier studies that had indicated that beta-ketoadipate does not permeate the membrane of wild-type P. putida cells. Contradictions between the former investigations and the present analysis are due primarily to the relatively high concentrations of substrate used in the earlier experiments. The existence of the beta-ketoadipate uptake system indicates that beta-ketoadipate may exist as a selective nutrient in the natural niche of P. putida and may play a determinative role in the evolution of induction mechanisms that are characteristic of fluorescent pseudomonads.
Extracellular and protease-released pullulanases
Authors: Brandt C J , Catley B J , Awad W M
Published: January 2020

The extracellular form of pullulanase (EC 3.2.1.41) from Klebsiella aerogenes has been purified to homogeneity by successive chromatography through diethylaminoethyl-cellulose, Sephadex G-200, and 1,6-diaminohexane-Sepharose. In addition, the cell-bound form of pullulanase has been released by the action of a serine endopeptidase obtained from Pronase and purified to apparent homogeneity. Protease-released pullulanase has a slightly larger molecular weight and a specific activity over twice that of the extracellular protein. The properties of each of these forms of pullulanase have been compared with those reported for the detergent-released form. Each form has different features as examined by amino acid composition, specific activity, molecular weight, or inhibition pattern, which distinguish it from the other pullulanases. It is hypothesized that a single gene product consisting of a single polypeptide chain generates these different enzyme forms after selective cleavages by endogenous or applied proteases.
Construction of intergeneric hybrids using bacteriophage P1CM: transfer of the Klebsiella aerogenes ribitol dehydrogenase gene to Escherichia coli
Authors: Rigby P W , Gething M J , Hartley B S
Published: January 2020

Study of many of the interesting properties of Klebsiella aerogenes is limited by the lack of a well-characterized genetic system for this organism. Our investigations of the evolution of the enzyme ribitol dehydrogenase (EC 1.1.1.56) in K. aerogenes would be greatly facilitated by the availability of such a system, and we here report two approaches to developing one. We have isolated mutants sensitive to the coliphage P1, which will efficiently tranduce genetic markers between such sensitive strains and which will thus make detailed mapping studies possible. Derivatives of K. aerogenes lysogenic for P1 can be readily isolated by using the specialized transducing particle P1CMclr100. Bacteria lysogenic for this phage are chloramphenicol resistant and temperature sensitive. Phage particles produced by temperature induction of such lysogens can be used to transfer K. aerogenes genes to the natural host of P1 phage. Escherichia coli. We have used this method to prepare derivatives of E. coli K-12 carrying the K. aerogenes genes conferring the ability to metabolize the pentitols ribitol and D-arabitol. We have shown that these E. coli-K. aerogenes hybrids synthesize a ribitol dehydrogenase with the properties of the K. aerogenes enzyme and have mapped the position of the transferred gene on the E. coli chromosome. The ramifications of this methodology are discussed.
Discontinuous synthesis of lac messenger ribonucleic acid in a mutant of Escherichia coli with a new lac promoter
Authors: Bruenn J , Kane W
Published: January 2020

A mutant of Escherichia coli with a new promoter for the lac operon exhibits dramatic discontinuities in the synthesis of lac messenger ribonucleic acid after induction. These discontinuities immediately precede similar discontinuities in the synthesis of beta-galactosidase. The discontinuous synthesis of beta-galactosidase persists after addition of rifampin.
Activator Specificity of Pyruvate Kinase from Lactic Streptococci
Authors: Thomas Terence D.
Published: January 2020

The pyruvate kinase from lactic streptococci was activated by 20 structurally dissimilar sugar phosphates and glycolytic intermediates. Nine compounds were more effective activators than fructose-1,6-diphosphate.
Effect of deoxyribonucleic acid replication inhibitors on bacterial recombination
Authors: Canosi U , Siccardi A G , Falaschi A , Mazza G
Published: January 2020

Two inhibitors of replicative deoxyribonucleic acid (DNA) synthesis, nalidixic acid (NAL) and 6-(p-hydroxyphenylazo)-uracil (HPUra), showed different effects on genetic recombination and DNA repair in Bacillus subtilis. Previous work (Pedrini et al., 1972) showed that NAL does not interfere with the transformation process of B. subtilis. The results reported in this work demonstrated that the drug was also without effect on the transfection by SPP1 or SPO-1 phage DNA (a process that requires a recombination event). The drug was also ineffective on the host cell reactivation of ultraviolet-irradiated SPP1 phage, as well as on transfection with ultraviolet-irradiated DNA of the same phage. HPUra instead markedly reduced the transformation process, as well as transfection, by SPO-1 DNA, but it did not affect the host cell reactivation of SPO-1 phage. In conclusion, whereas the NAL target seems to be specific for replicative DNA synthesis, the HPUra target (i.e., the DNA polymerase III of B. subtilis) seems to be involved also in recombination, but not in the excision repair process. The mutations conferring NAL and HPUra resistance used in this work were mapped by PBS-1 transduction.
Sequence of molecular events involved in induction of allophanate hydrolase
Authors: Bossinger J , Cooper T G
Published: January 2020

Addition of urea to an uninduced culture of Saccharomyces at 22 C results in appearance of allophanate hydrolase activity after a lag of 12 min. We have previously demonstrated that both ribonucleic acid (RNA) and protein synthesis are needed for this induction to occur. To elucidate the time intervals occupied by known processes involved in induction, temperature-sensitive mutants defective in messenger RNA transport from nucleus to cytoplasm (rna1) and in protein synthesis initiation (prt1) were employed along with an RNA polymerase inhibitor in experiments that measure cumulative synthetic capacity to produce allophanate hydrolase. These measurements identify the time within the lag period at which each of the above processes is completed. We observed that RNA synthesis, rna1 gene product function, and protein synthesis initiation are completed at 1 to 1.5, 4, and 9 to 10 min, respectively.
Effect of arginine on the stability and size of argECBH messenger ribonucleic acid in Escherichia coli
Authors: Krzyzek R A , Rogers P
Published: January 2020

The chemical stability of argECBH messenger ribonucleic acid (mRNA) produced by Escherichia coli was found to be unaltered during steady-state repression by arginine. During extreme arginine deprivation, the increase in argECBH mRNA stability was related to general effects of amino acid starvation on mRNA stability. Thus a mechanism whereby argECBH gene expression is regulated by altering the decay rate of this mRNA is not consistent with our data. Sucrose gradient analysis followed by hybridization revealed that both heavy (14S) and light (8S) components of argECBH mRNA were produced by cells of E. coli K-12 grown without arginine, whereas predominantly light (8S) mRNA was produced by cells grown with arginine. A functional argR gene and the EC portion of the argECBH cluster were found essential for the arginine restriction of heavy-mRNA production. Experiments suggest that light argECBH mRNA did not arise from heavy message, and 8u% of both light and heavy mRNA was found bound to ribosomes. The data appear most consistent with the notion that a second site of control by arginine regulates the amounts of light and heavy arginine mRNA in the cell either by early termination of transcription or by endonucleolytic processing. Consideration of these data in conjunction with those of the accompanying report (Krzyzek and Rogers, 1976) permits the tentative conclusion that light argECBH mRNA is not translated into active enzymes and is thus responsible for the discrepancy between the high content of hybridizable mRNA and low rates of enzyme synthesis found during arginine repression.
Isolation and characterization of the specialized transducing bacteriophages phi80dargF and lambdah80cI857 dargF: specific cleavage of arginine transducing deoxyribonucleic acid by the endonucleases EcoRI and SmaR
Authors: James P M , Sens D , Natter W , Moore S K , James E
Published: January 2020

The directed transposition of argF to the tonB locus of the Escherichia coli chromosome and the subsequent isolation of the specialized transducing phage phi80dargF is described. The structure of this phage has been has been determined. A hybrid lambdah80cI857dargF phage has been constructed. Deoxyribonucleic acid isolated from these and their parent bacteriophages has been specifically cleaved by the endonucleases EcoRI and SmaR; the unique deoxyribonucleic acid fragments thus obtained have been resolved and analyzed by electrophoresis in agarose gel.
Active Subunits of
Escherichia coli
Glutamate Synthase
Authors: Mäntsälä Pekka , Zalkin Howard
Published: January 2020

The large and small subunits of
Escherichia coli
glutamate synthase were isolated. The small subunit catalyzes the NH
3
-dependent synthesis of glutamate. The large subunit exhibits glutaminase activity.
Repair of ultraviolet light-induced damage in Micrococcus radiophilus, an extremely resistant microorganism
Authors: Lavin M F , Jenkins A , Kidson C
Published: January 2020

Repair of ultraviolet radiation damage was examined in an extremely radioresistant organism, Micrococcus radiophilus. Measurement of the number of thymine-containing dimers formed as a function of ultraviolet dose suggests that the ability of this organism to withstand high doses of ultraviolet radiation (20,000 ergs/mm2) is not related to protective screening by pigments. M. radiophilus carries out a rapid excision of thymine dimers at doses of ultraviolet light up to 10,000 ergs/mm2. Synthesis of deoxyribonucleic acid is reduced after irradiation, but after removal of photodamage the rate approaches that in unirradiated cells. A comparison is drawn with Micrococcus luteus and M. radiodurans. We conclude that the extremely high resistance to ultraviolet irradiation in M. radiophilus is at least partly due to the presence of an efficient excision repair system.
Formation in the dark, of virus-induced deoxyribonuclease activity in Anacystis nidulans, an obligate photoautotroph
Authors: Udvardy J , Sivok B , Borbely G , Farkas G L
Published: January 2020

In Anacystis nidulans, upon infection with cyanophage AS-1, after a lag period of 1 h the level of deoxyribonuclease (DNase) activity increaded rapidly up to 15- to 20-fold in 4 to 5 h in the light. In contrast, the ribonuclease and phosphomonoesterase activities increased significantly only 4 to 5 h after infection, i.e. as late as 1 h prior to lysis. In complete darkness, the nuclease levels remained unaltered. However, when the infected cells were exposed to light for 1 or 2 h after infection, the DNase level increased essentially to the same extent in the dark as in continuous light, although the complete replication cycle of the virus was impaired in the dark and cells lysed only in the continuously illuminated cultures. Inhibition of photosystem II with 3-(3,4-dichlorophenyl)-1-dimethylurea during the early illumination period strongly decreased the subsequent, infection-dependent increase in DNase activity in the dark. The virus-induced increase in DNase activity was also inhibited by chloramphenicol. The data suggest that, in spite of the obligate photoautotrophic nature of A. nidulans, dark metabolism is able to support fully the formation of some specific proteins if the triggering of their synthesis takes place in light.
Clo DF13 plasmid deoxyribonucleic acid-directed in vitro synthesis of biologically active cloacin DF13 and clo DF13 immunity protein
Authors: Konings R N , Andreoli P M , Veltkamp E , Nijkamp H J
Published: January 2020

Clo DF13 plasmid deoxyribonucleic acid (DNA) was used as a template to direct transcription and translation in a DNA-dependent cell-free system prepared from Escherichia coli. Analysis of the invitro products on sodium dodecylsulfate-polyacrylamide gels revealed that Clo DF13 DNA directs the synthesis of at least 10 polypeptides ranging in molecular weight from approximately 7,000 to 70,000. Two of these polypeptides could be identified, with respect to their physiochemical and biological characteristics, as the products of the Clo DF13 genes coding for cloacin DF13 and Clo DF13 immunity protein. These results confirm previous findings, obtained which Clo DF13-harbouring minicells of E. coli, that the structural tenes for the latter proteins residue on the Clo DF13 genome.
Mutants in three genes affecting transport of magnesium in Escherichia coli: genetics and physiology
Authors: Park M H , Wong B B , Lusk J E
Published: January 2020

Mutants in three genes affecting two Mg2+ transport systems are described. System I, for which Co2+, Mn2+, and Mg2+ are substrates, is inactive in corA mutants corB mutants express system I after growth on high (10 mM) Mg2+ but not low (0.1 mM) Mg2+. Both corA and corB mutants are resistant to Co2+ or Mn2+. corA mutants are sensitive to CA2+. Transport system II is specific for Mg2+ and is repressed by growth on 10 mM Mg2+. mgt mutations inactivate system II. Growth on mgt mutants in normal except on very low (1 muM) concentrations of Mg2+, corA mgt strains exhibit no high-affinity, energy-dependent transport of Mg2+ and require 10 mM Mg2+ for optimal growth. The three genes are not linked. The corA locus is contransducible with ilv at 75 min, corB is cotransducible with pyrB at 85 min, and mgt is cotransducible with malB and mel at 81 min on the genetic map.
Effect of araC gene product on catabolite repression in the L-arabinose regulon
Authors: Heffernan L , Wilcox G
Published: January 2020

The araCi protein differs in stability from araC+ protein and alters the concentration of cyclic adenosine-3', 5'-monophosphate required to maximally stimulate L-arabinose isomerase synthesis in an in vitro protein-synthesizing system.
Altered properties of deoxyribonucleic acid polymerase I isolated from the membrane of bacteriophage T4-infected Escherichia coli
Authors: Majumdar C , Dewey M , Frankel F R
Published: January 2020

Previous studies have shown that a large fraction of the host cell deoxyribonucleic acid (DNA) polymerase I (EC 2.7.7.7) becomes associated with the cell membrane shortly after infection with bacteriophages T4 and T7. The present investigation of the bound enzyme revealed that the polymerase activity can be eluted from the membrane with chelating agents, and that the material thus obtained shows many properties that distinguish it from purified DNA polymerase I. These include its chromatographic behavior, sedimentation rate, sensitivity to anti-DNA polymerase I antiserum, and activity with synthetic and natural DNA primers. Several of these physical and biological parameters were shown to revert slowly during storage to those exhibited by the purified enzyme. Efforts to determine whether the unusual properties of the membrane enzyme resulted from its association with DNA failed to support that possibility. These observations suggest that either the cause or the result of membrane binding of DNA polymerase I is a transient change in conformation or structure of the enzyme, with a resultant change in its enzymatic activity.
Regulation of beta-glucuronidase synthesis in Escherichia coli K-12: pleiotropic constitutive mutations affecting uxu and uidA expression
Authors: Novel M , Novel G
Published: January 2020

Among the beta-glucuronidase (UID)-constitutive mutants obtained by growth on methyl-beta-D-galacturonide, some strains are also derepressed for the two enzymes of the uxu operon: mannonate oxidoreductase (MOR) and mannonate hydrolyase (HLM). By conjugation and transduction experiments, two distinct constitutive mutations were separated in each pleiotropic mutant strain. One of them was specific for uidA gene expression and was characterized as affecting either uidO or uidR sites. The second type of mutation was mapped close to the uxu operon and was found to be responsible for the pleiotropic effect revealed in the primary mutants: after separation such a mutation still fully derepresses MOR and HLM synthesis but weakly derepresses UID synthesis. The pleiotropic effect of this mutation was maintained even though the activity of the structural genes was altered. This rules out the occurrence of an internal derepressing interaction between these enzymes. In merodiploid strains, uxu-linked constitutive mutations were recessive to the wild-type allele, suggesting that these mutations could affect a regulatory gene. The uxuR gene is probably a specific regulatory gene for a very close operon, uxu. Moreover, it has a weak effect on uidA expression. Thus, UID synthesis would be negatively controlled through the activity of two repressor molecules that are synthesized by two distinct regulatory genes, uidR and uxuR. These two repressing factors are antagonized, respectively, by phenyl-thio-beta-D-glucuronide and mannonic amide and could cooperate in a unique repression/induction control over uidA expression. Constitutive mutations affecting the control sites of uidA gene probably characterize two distinct attachment sites in the operator locus for each of the repressor molecules.
Comparative study of R1-specific chromosomal transfer in Escherichia coli K-12 and salmonella typhimurium LT2
Authors: Hedén L O , Meynell E
Published: January 2020

High-frequency transfer of the chromosomal trp region by R1 observed in Escherichia coli (Pearce and Meynell, 1968) also occurs in Salmonella typhimurium. The reaction is recA independent in both species. The origin of transfer lies within a segment of the chromosome that is inverted in S. typhimurium relative to E. coli, and thus transfer occurs in a different direction in the two species. The character of R1 that is responsible, known as Tfa+, may be lost without affecting other properties of the R factor such as its own transfer.
Uptake of branched-chain alpha-keto acids in Bacillus subtilis
Authors: Goldstein B J , Zahler S A
Published: January 2020

Bacillus subtilis has a constitutive system for the uptake of alpha-keto-beta-methylvalerate, alpha-ketoisovalerate, and (probably) alpha-ketoisocaproate. A mutation, kauA1, which blocks the uptake of alpha-keto-beta-methylvalerate and alpha-ketoisovalerate, is located between metB and citK on the B. subtilis chromosome.
Selection for citrate synthase deficiency in icd mutants of Escherichia coli
Authors: Lakshmi T M , Helling R B
Published: January 2020

Cultures of isocitrate dehydrogenase-deficient (icd) mutants were overgrown by double mutants (icd glt) lacking citrate synthase activity also. The icd mutants grew more slowly than wild-type cells or the double mutants because they accumulated an inhibitory metabolite (possibly citrate). Intracellular citrate levels were several hundred-fold higher in icd cells than in wild-type or icd glt cells. Final growth yields of the wild type and the icd mutant on limiting glucose were equivalent and greater than the growth yield of icd glt double mutants. The icd gene mapped between 60 and 74 min. icd mutants were resistant to nalidixic acid, but glt and icd glt mutants and wild-type cells were sensitive, indicating that resistance results from accumulation of isocitrate, citrate, or a derivative of these compounds.
Role of Rec pathways on sensitivity of Escherichia coli to near-ultraviolet and visible light
Authors: Gopalakrishna K , Bhattacharjee S K
Published: January 2020

In Escherichia coli lack of the RecRC or RecF pathway is found to cause sensitivity to near-ultraviolet and visible light. Resistance to this light is restored in the RecBC-defective strain carrying either the sbcB (Rec+) or xonA (Rec-) mutation. The sensitivity, therefore, is not found to correlate with the degree of recombination proficiency as measured by genetic crosses.
Hydrogenase and ribulose diphosphate carboxylase during autotrophic, heterotrophic, and mixotrophic growth of scotochromogenic mycobacteria
Authors: Park S S , DeCicco B T
Published: January 2020

Two key autotrophic enzyme systems, hydrogenase and ribulose diphosphate carboxylase, were examined in Mycobacterium gordonae and two other chemolithotrophic, scotochromogenic mycobacteria under different cultural conditions. In all three organisms both enzymes were inducible and were produced in significant levels only in the presence of the specific substrate, hydrogen or carbon dioxide. M. gordonae exhibited increased growth rates and yields, indicating mixotrophic growth, in the presence of a number of single organic substrates, including acetate, pyruvate, glucose, fructose, and glycerol. In contrast to other aerobic hydrogen autotrophs, the presence of either acetate or pyruvate did not repress ribulose diphosphate carboxylase, and mixotrophic growth was rapid with these substrates. In the absence of carbon dioxide, growth in glycerol medium under an atmosphere of hydrogen and oxygen was severely inhibited, even with cells preadapted to heterotrophic growth on glycerol. Cyclic adenosine monophosphate was not effective in inducing hydrogenase or carboxylase in heterotrophic, mixotrophic, or hydrogen-inhibited cultures.