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C A Reddy

Publications and source records attributed to C A Reddy.

At least 73 records · Page 4Linked to original sources
PubMed1983

Construction of a new yeast cloning vector containing autonomous replication sequences from Candida utilis.

DNA sequences from the Candida utilis genome which, when cloned into a yeast integration plasmid (YIp5), confer on YIp5 the ability to replicate autonomously in Saccharomyces cerevisiae are described. Several recombinant plasmids which transform S. cerevisiae YNN27 to Ura3+ with an efficiency of 2 X 10(3) transformants per microgram of DNA were obtained. One of the recombinant plasmids, pHMR22 (6.6 kilobases) contains ars (autonomous replication sequence), which is homologous with two different DNA fragments of the C. utilis genome but has no detectable homology to total DNA from Candida albicans, Pachysolen tannophilus, or S. cerevisiae. Restriction and subcloning analyses of pHMR22 showed that Sau3A destroys the functions of cloned ars whereas there are no BamHI, PstI, SalI, HindIII, EcoRI, or PvuII sites in the region of ars which is required for its functional integrity. Thus, pHMR22 appears to be a useful vector for cloning desired genes in S. cerevisiae.

Base Sequence↗
PubMed1982

The involvement of hydroxyl radical derived from hydrogen peroxide in lignin degradation by the white rot fungus Phanerochaete chrysosporium.

The possible involvement of hydrogen peroxide (H2O2)-derived hydroxyl radical (.OH) in lignin degradation ([14C]lignin leads to 14CO2) by Phanerochaete chrysosporium was investigated. When P. chrysosporium was grown in low nitrogen medium (2.4 mM N), an increase in the specific activity for H2O2 production in cell extracts was observed to coincide with the appearance of ligninolytic activity and both activities appeared after the culture entered stationary phase. The production of .OH in ligninolytic cultures of P. chrysosporium was demonstrated by alpha-keto-gamma-methiolbutyric acid-dependent formation of ethylene. Hydrogen peroxide-dependent .OH formation was also shown in cell extracts of ligninolytic cultures. The radical species was demonstrated to be .OH by the .OH-dependent hydroxylation of p-hydroxybenzoic acid to form protocatechuic acid and by using 5,5-dimethyl-1-pyrroline-N-oxide and detecting the production of the nitroxide radical of 5,5-dimethyl-1-pyrroline-N-oxide by EPR. These reactions were inhibited by .OH-scavenging agents and were stimulated when azide was added to inhibit endogenous catalase. Lignin degradation by P. chrysosporium was markedly suppressed in the presence of the .OH-scavenging agents mannitol, benzoate, and the nonspecific radical scavenging agent butylated hydroxytoluene. The above results indicate that .OH derived from H2O2 is involved in lignin biodegradation by P. chrysosporium.

Agaricales↗
PubMed1982

Ethylene production from alpha-oxo-gamma-methylthiobutyric acid is a sensitive measure of ligninolytic activity by Phanerochaete chrysosporium.

Production of hydroxyl radical in ligninolytic cultures was determined by measuring the alpha-oxo-gamma-methylthiobutyric acid-dependent production of ethylene gas. The results showed that the pattern of ethylene production was very similar to that of ligninolytic activity [[14C]lignin leads to 14CO2). Furthermore, nutritional parameters, which are known to affect ligninolytic activity, affected OH.-radical-dependent ethylene production in a similar fashion. The results indicate that assay for ethylene production from alpha-oxo-gamma-methylthiobutyric acid is a simple and sensitive measure of ligninolytic activity by Phanerochaete chrysosporium.

Basidiomycota↗
PubMed1982

Physiological and nutritional features of Corynebacterium pyogenes.

Growth and acid metabolic products were similar when Corynebacterium pyogenes was grown aerobically or anaerobically in a serum-free medium (SFM). This indicated that C. pyogenes obtains energy for growth primarily by fermentative metabolism even under aerobic growth conditions. Growth yield was reduced by 90% in SFM minus glucose, 50% in SFM minus NaHCO3, 90% in SFM minus yeast extract, 100% in SFM minus Trypticase and yeast extract, and 30% in SFM minus haemin or Trypticase. Growth was not detectable when a known mixture of amino acids, vitamins, and nucleic acid bases were substituted for Trypticase and yeast extract in SFM; addition to the latter medium of a peptide source such as Trypticase or casitone supported good growth of the organism. When NaHCO3 was omitted from SFM and dissolved CO2 in the medium was rigorously excluded, growth was undetectable indicating that C. pyogenes has an obligate requirement for CO2 for growth. Succinate, formate and acetate were the major fermentation products in SFM, whereas in SFM minus HCO-3 or haemin, lactate was the major product and only small quantities of other acids accumulated.

Aerobiosis↗
PubMed1982

Ultrastructural Localization of Hydrogen Peroxide Production in Ligninolytic Phanerochaete chrysosporium Cells.

Previous studies have shown that the hydroxyl radical derived from hydrogen peroxide (H(2)O(2)) is involved in lignin degradation by Phanerochaete chrysosporium. In the present study, the ultrastructural sites of H(2)O(2) production in ligninolytic cells of P. chrysosporium were demonstrated by cytochemically staining cells with 3,3'-diaminobenzidine (DAB). Hydrogen peroxide production, as evidenced by the presence of oxidized DAB deposits, appeared to be localized in the periplasmic space of cells from ligninolytic cultures grown for 14 days in nitrogen-limited medium. When identical cells were treated with DAB in the presence of aminotriazole, periplasmic deposits of oxidized DAB were not observed, suggesting that the deposits resulted from the H(2)O(2)-dependent peroxidatic oxidation of DAB by catalase. Cells from cultures grown for 3 or 6 days in nitrogen-limited medium or for 14 days in nitrogen-sufficient medium had little ligninolytic activity and low specific activity for H(2)O(2) production and did not contain periplasmic oxidized DAB deposits. The results suggest that in cultures grown in nitrogen-limited medium, there is a positive correlation between the occurrence of oxidized DAB deposits, the specific activity for H(2)O(2) production in cell extracts, and ligninolytic activity.

Journal Article↗
PubMed1982

Nutritional and metabolic features of Eubacterium suis.

We studied the nutritional and metabolic features of Eubacterium suis, an anaerobic animal pathogen that causes cystitis and pyelonephritis in pigs. Peptone-yeast extract-starch (PYS) medium, which contained Trypticase (BBL Microbiology Systems), yeast extract, starch, minerals, cysteine, and sodium carbonate, was shown to support excellent growth of this organism (absorbance at 600 nm = 1.8). Growth was considerably less (absorbance at 600 nm = 0.6) when the starch in the medium was replaced by maltose. Formate, acetate, and ethanol were the major products of fermentation of starch or maltose. The organism appears to require a fermentable carbohydrate for growth since the deletion of starch from PYS resulted in a negligible amount of growth. Growth decreased by approximately 20% when CO2 was rigorously excluded from PYS minus Na2CO3. The deletion of only yeast extract from PYS resulted in a decrease in growth of about 75%, and the simultaneous deletion of both yeast extract and Trypticase resulted in negligible growth. When the yeast extract in PYS was replaced by a defined mixture of purine and pyrimidine bases, vitamins, and amino acids, growth was greater than or equal to 80% that observed in PYS. The deletion of Trypticase from this medium resulted in no detectable growth, suggesting a possible peptide requirement for E. suis growth. Good growth (absorbance at 600 nm = 1.4) was obtained when adenine and uracil were substituted for the mixture of purine and pyrimidine bases in modified PYS; the substitution of pyridoxal, riboflavin, and nicotinic acid for the vitamin mixture gave comparable growth. The nutritional requirement of E. suis apparently reflect the fact that the organism adapts to its natural niche by doing away with certain biosynthetic capabilities which it does not seem to require.

Carbon Dioxide↗
PubMed1982

Nutritional requirements of Corynebacterium pyogenes.

The nutritional requirements of Corynebacterium pyogenes (strains C100, 5, and 1909), a commonly encountered animal pathogen, were determined in this study. A semidefined medium (SDM) containing glucose, HCO3-, hemin, charcoal-treated Trypticase, and a defined mixture of purines and pyrimidines, amino acids, and minerals which supported optimal growth of C. pyogenes was employed in all nutritional studies. Adenine and uracil were required for optimal growth of strains 5 and C100 but were not required for strain 1909. Riboflavin and nicotinic acid were required for good growth of all three strains; biotin and thiamin were stimulatory but did not appear to be required for growth. Hemin and NaHCO3 were stimulatory for growth, whereas lipoic acid and Tween 80 were neither stimulatory nor required for growth. The replacement of Trypticase with a specific peptide fraction (obtained by fractionation of Trypticase on Sephadex G-25) rich in dipeptides gave growth comparable to that in SDM, indicating a peptide requirement for the growth of C. pyogenes. It was of considerable interest that growth comparable to that in SDM was obtained when Trypticase was replaced by inositol (1 microgram/ml of SDM).

Amino Acids↗
PubMed1980

Chemically defined growth medium for Corynebacterium pyogenes.

A chemically defined medium and a relatively simple semidefined medium (SDM) which allow good growth of Corynebacterium pyogenes, a nutritionally fastidious animal pathogen, were described. The SDM contained glucose, trypticase, yeast extract, hemin, minerals, cysteine x HCl, and NaHCO3. To obtain a chemically defined medium, yeast extract in SDM was replaced with a defined mixture of nucleic acid bases, vitamins, amino acids, and trace minerals, and trypticase was replaced by myo-inositol (1 microgram/ml of medium).

Animals↗
PubMed1978

Electron transport phosphorylation coupled to fumarate reduction by H2- and Mg2+-dependent adenosine triphosphatase activity in extracts of the rumen anaerobe Vibrio succinogenes.

Vibrio succinogenes, an anaerobic bacterium, obtains its energy for growth from H2 or formate oxidation coupled to the reduction of fumarate to succinate. Membrane preparations have been obtained from this organism that catalyze the synthesis of ATP during H2 oxidation coupled to fumarate reduction. Esterification of orthophosphate is dependent on electron transfer, as evidenced by the requirement for both H2 and fumarate. Phosphorylation is also dependent on ADP and is destroyed by boiling the membrane preparations. H2 utilized for fumarate reduction and succinate formed are stoichiometric. The phosphorylation is markedly uncoupled by pentachlorophenol and gramicidin, but to a lesser extent by dinitrophenol and methyl viologen. 2-n-Heptyl-4-hydroxyquinoline-N-oxide causes severe inhibition of H2 oxidation as well as phosphorylation, but oligomycin or antimycin A has no demonstrable effect. Among several electron acceptors tested, significant phosphorylation is observed only with fumarate. A Mg2+-dependent adenosine triphosphatase activity is present in both the membrane and soluble protein fractions. Highest activity is obtained with ATP as the substrate, and considerably less activity is obtained with other nucleoside triphosphates. The possibility that phosphorylation during "fumarate respiration" may play an important physiological role in the growth of many anaerobic and facultatively anaerobic bacteria is discussed.

Adenosine Triphosphatases↗
PubMed1978

Value of acid metabolic products in identification of certain corynebacteria.

Acid metabolic products of 23 strains of human and animal pathogenic corynebacteria, representing eight different species, were determined by gas chromatography. The results showed that the species examined were metabolically heterogeneous and could be presumptively identified based on the acid products produced. Corynebacterium equi did not produce any acids; C. renale produced lactate; and C. pyogenes produced major amounts of lactate, variable amounts of acetate, and minor amounts of succinate and pyruvate. C. kutscheri produced propionate and lactate as major products and pyruvate and oxalacetate as minor products. C. diphtheriae and C. pseudotuberculosis produced major amounts of propionate, acetate, and formate. In addition, C. pseudotuberculosis produced major amounts of pyruvate and minor amounts of succinate, lactate, and oxalacetate, whereas C. diphtheriae strains produced minor but variable amounts of lactate, succinate, fumarate, pyruvate, and oxalacetate. C. bovis produced aicd products similar to those of C. pyogenes but was readily distinguishable from the latter by the lack of hemolysis on blood agar, colony morphology, catalase reaction, and biochemicals. C. suis characteristically produced major amounts of ethanol, acetate, and formate and minor amounts of lactate and succinate but no propionate.

Carboxylic Acids↗
PubMed1977

Amino acid profiles and presumptive nutritional assessment of single-cell protein from certain lactobacilli.

The amino acid profiles, modified essential amino acid (MEAA) indexes, and in vitro pepsin digestibilities were determined for single-cell protein (SCP) from certain industrially important lactobacilli. For the three parameters examined, substantial differences were seen between different Lactobacillus species and between strains with a given species. SCP from all of the lactobacilli examined appeared relatively high in MEAA indexes and pepsin digestibility. SCP from L. acidophilus 3205 and L. fermenti 3954 had the highest MEAA indexes, whereas L. bulgaricus 2217 and L. thermophilus 3863 had the highest percentage of digestible crude protein. SCP from L. plantarum strains had the lowest MEAA indexes. The essential amino acid compositions of SCP from different lactobacilli appear comparable to that of Food and Agriculture Organization reference protein and SCP from other sources.

Amino Acids↗
PubMed1977

Growth of desulfovibrio in lactate or ethanol media low in sulfate in association with H2-utilizing methanogenic bacteria.

In the analysis of an ethanol-CO(2) enrichment of bacteria from an anaerobic sewage digestor, a strain tentatively identified as Desulfovibrio vulgaris and an H(2)-utilizing methanogen resembling Methanobacterium formicicum were isolated, and they were shown to represent a synergistic association of two bacterial species similar to that previously found between S organism and Methanobacterium strain MOH isolated from Methanobacillus omelianskii. In lowsulfate media, the desulfovibrio produced acetate and H(2) from ethanol and acetate, H(2), and, presumably, CO(2) from lactate; but growth was slight and little of the energy source was catabolized unless the organism was combined with an H(2)-utilizing methanogenic bacterium. The type strains of D. vulgaris and Desulfovibrio desulfuricans carried out the same type of synergistic growth with methanogens. In mixtures of desulfovibrio and strain MOH growing on ethanol, lactate, or pyruvate, diminution of methane produced was stoichiometric with the moles of sulfate added, and the desulfovibrios grew better with sulfate addition. The energetics of the synergistic associations and of the competition between the methanogenic system and sulfate-reducing system as sinks for electrons generated in the oxidation of organic materials such as ethanol, lactate, and acetate are discussed. It is suggested that lack of availability of H(2) for growth of methanogens is a major factor in suppression of methanogenesis by sulfate in natural ecosystems. The results with these known mixtures of bacteria suggest that hydrogenase-forming, sulfate-reducing bacteria could be active in some methanogenic ecosystems that are low in sulfate.

Acetates↗
PubMed1977

Hydrogenase activity and the H2-fumarate electron transport system in Bacteroides fragilis.

Hydrogenase activity and the H(2)-fumarate electron transport system in a carbohydrate-fermenting obligate anaerobe, Bacteroides fragilis, were investigated. In both whole cells and cell extracts, hydrogenase activity was demonstrated with methylene blue, benzyl viologen, flavin mononucleotide, or flavin adenine dinucleotide as the electron acceptor. A catalytic quantity of benzyl viologen or ferredoxin from Clostridium pasteurianum was required to reduce nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate with H(2). Much of the hydrogenase activity appeared to be associated with the soluble fraction of the cell. Fumarate reduction to succinate by H(2) was demonstrable in cell extracts only in the presence of a catalytic quantity of benzyl viologen, flavin mononucleotide, flavin adenine dinucleotide, or ferredoxin from C. pasteurianum. Sulfhydryl compounds were not required for fumarate reduction by H(2), but mercaptoethanol and dithiothreitol appeared to stimulate this activity by 59 and 61%, respectively. Inhibition of fumarate reduction by acriflavin, rotenone, 2-heptyl-4-hydroxyquinoline-N-oxide, and antimycin A suggest the involvement of a flavoprotein, a quinone, and cytochrome b in the reduction of fumarate to succinate. The involvement of a quinone in fumarate reduction is also apparent from the inhibition of fumarate reduction by H(2) when cell extracts were irradiated with ultraviolet light. Based on the evidence obtained, a possible scheme for the flow of electrons from H(2) to fumarate in B. fragilis is proposed.

Acriflavine↗
PubMed1977

Deoxyribonucleic acid base composition of certain species of the genus Bacteroides.

The moles percent guanine plus cytosine content of the DNA (% G + C) of 15 Bacteroides strains representing six species was determined. One group, including three strains of Bacteroides ruminicola, two strains of B. melaninogenicus, two strains of B. succinogenes, and one strain of B. oralis (JI), had a % G + C of 47.6--50.3 and a second group including two strains of B. amylophilus, four strains of B. fragilis, and one strain of B. succinogenes had a lower % G + C of 40.3--42.7. The taxonomic relationships among these bacteroides species were discussed.

Animals↗
PubMed1976

Anaerobic bacteria from the large intestine of mice.

Anaerobic bacteria from the colon of laboratory mice were enumerated and isolated using strict anaerobic techniques. Direct microscopic counts revealed 4.4 X 10(10) organisms in each gram (wet weight) of colon contents. Actual cultural counts averaged 3.2 X 10(10) organisms, which was 73% of the direct microscopic count. The tentatively identified genera were Bacteroides, Eubacterium, Fusobacterium, Lactobacillus, Peptostreptococcus, and Propionibacterium. Strains of Fusobacterium, Lactobacillus, Peptostreptococcus, and Propionibacterium were biochemically homogeneous. Strains of Bacteroides and Eubacterium, on the other hand, were biochemically heterogeneous and were subdivided into several distinct groups. The data indicate that many of the isolates are different from previously described species of the respective genera and may belong to new species.

Anaerobiosis↗