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PubMed1993

Extracellular fibrinogenolytic enzyme of Aspergillus fumigatus: substrate-dependent variations in the proteinase synthesis and characterization of the enzyme.

To get a better understanding of the role of the previously reported fibrinogenolytic enzyme of Aspergillus fumigatus, we investigated the in vitro conditions of enzyme synthesis and attempted to characterize it. Modification of the nitrogen source did not influence the extracellular serine-proteinase profile, but resulted in important quantitative differences in the yields in batch cultures. The enzyme synthesis appeared to be an inducible phenomenon in A. fumigatus since it was initiated exclusively in the presence of proteins or protein hydrolysate. Free amino acids or inorganic nitrogen compounds could not promote significant enzyme production. Moreover, peptone at a concentration of 0.1% appeared to be the best inducer of enzyme synthesis. Conversely, modification of the carbon source did not affect fungal growth or enzyme synthesis. However, the production of chymotrypsin was highly sensitive to the carbohydrate level in the culture medium and, with peptone as nitrogen source, highest yields were obtained in the presence of 0.3 or 0.5% glucose. Culture filtrates of A. fumigatus CBS 113.26 grown with peptone or nitrate as nitrogen source were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Comparison of the protein patterns suggested for the proteinase a molecular mass of 33 kDa which was confirmed by chromatographic purification of the enzyme through (N alpha-CBZ)-D-phenylalanine agarose.

Amino Acid Sequence↗

PubMed1976

Endosymbionts of Sitodrepa panicea.

Morphological and physiological characteristics of seven strains of yeast-like symbionts isolated from Sitodrepa panicea justify their inclusion into the genus Torulopsis as a new species: T. buchnerii. The symbiotic relationship is mutually beneficial: the symbionts obtain some nitrogenous compounds and carbohydrates, such as proline and trehalose from the host's hemolymph, and synthesize and make available to the host all the essential amino acids and vitamins, except biotin.

Animals↗

PubMed1998

Transcriptional control of the glnD gene is not dependent on nitrogen availability in Escherichia coli.

Glutamine synthetase (GS) is one of the most important enzymes in the assimilation of nitrogenous compounds in Escherichia coli and related bacteria. For the control of its activity and biosynthesis, tricyclic cascades of uridylylation/deuridylylation of PII protein, adenylylation/deadenylylation of glutamine synthetase, and phosphorylation/dephosphorylation of Ntr1 are operating, where the regulation of uridylylation/deuridylylation by uridylyl transferase-uridylyl removing enzyme (UT-UR) (the product of the glnD gene) would play the ultimate nitrogen sensing role. However, the possible nitrogen-regulatable element in the upstream of the glnD gene has been debated. In the present experiment, we have cloned and sequenced the four minute regions of the Escherichia coli chromosome, where rpsB, map, glnD, and dapD genes have been identified in sequence. We could localize the transcriptional start site at seven nucleotides upstream of the translation initiation codon by primer extension analysis. The nitrogen dependency of the glnD gene has been analyzed by Northern blot, RNase protection, and promoter-luciferase activity assays. These data suggested a constitutive expression of the glnD gene independent of nitrogen availability. From these results, it could be concluded that the ultimate nitrogen sensing device for the bacteria should be the UT-UR itself, through modulation of its activity in response to the nitrogen status rather than its biosynthetic mechanism.

Amino Acid Sequence↗

PubMed1988

Regulation of nitrogenase gene expression in anaerobic cultures of Anabaena variabilis.

Derepression of nitrogenase gene expression was studied at the mRNA and enzyme activity levels in anaerobic cultures of Anabaena variabilis 29413. Cells, previously grown with ammonium chloride, were incubated in the absence of fixed nitrogen compounds under an Ar atmosphere with dichlorophenyldimethyl-urea present to inhibit oxygen evolution. The appearance of nitrogenase mRNA (measured by dot blot hybridization analysis) and nitrogenase activity (measured as acetylene-reducing activity) was followed, and the cells were also observed by phase-contrast microscopy. Nitrogenase mRNA could be detected after 1.5 to 2.0 h of nitrogen starvation; enzyme activity appeared about 1 h later. Although enzyme activity increased for many hours, mRNA levels reached a steady state rapidly. Neither heterocysts nor proheterocysts formed under these conditions; however, the cells were observed to shrink and become chlorotic. When anaerobic, derepressed cultures were exposed to oxygen, nitrogenase mRNA levels decreased very rapidly.

Anaerobiosis↗

PubMed1975

[Effect of the ruminal amino nitrogen level on the nitrogen passage into the isolated rumen of sheep].

Eight trials were performed with two sheep to study the passage of ammonia nitrogen, urea nitrogen, and amino nitrogen into an isolated rumen at different amino nitrogen levels in rumen after a single intraruminal application of enzymatic casein hydrolyzate. After casein hydrolyzate application the level of amino nitrogen in rumen increased; consequently, the passage of ammonia to the isolated rumen rapidly decreased. The passage of nitrogen as urea and amino nitrogen to the isolated rumen is the same, both at a higher and a lower level of amino nitrogen in the rumen. The total quantity of urea and ammonia nitrogen (N-NH3 + N-urea) and amino nitrogen (N-NH3 + N-urea + amino-N) present in the isolated rumen shows a highly significant correlation with ammonia passage. This quantity reaches its maximum before the application of casein hydrolyzate to the rumen. An intensive drop occurs within one and two hours after application. The results of our study testify to the fact that the chemical composition of rumen content plays an important role in the ruminohepatic circulation of nitrogen, particularly when endogenic nitrogen passes into the rumen through rumen wall, and that the passage of nitrogen compounds from blood to the rumen is influenced not only by the concentration ratio between blood and rumen content but also by neurohumoral effects.

Amino Acids↗

PubMed1994

Effects of organic acids on heterotrophic nitrification by Alcaligenes faecalis OKK17.

Factors affecting heterotrophic nitrification by Alcaligenes faecalis OKK17, which was isolated from sewage sludge, were examined. Specific nitrifying activity increased as the pH increased up to 8.5. Most of the nitrogenous compounds (88%) in the culture supernatant were converted to hydroxylamine or nitrite at pH 9 but 87% of them remained as ammonium at pH 7. These results imply that the substrate for heterotrophic nitrification is ammonium and that the organism oxidizes ammonium to lower its toxic effect. Although the addition of acetate to a defined medium increased growth of the bacterium up to C/N = ca. 6, the accumulation of nitrification products almost paralleled the growth and the specific nitrifying activity decreased. Pyruvate and oxaloacetate increased the specific nitrifying activity six- to eightfold compared with the other organic acids examined, but the key enzyme activities in the glyoxylate cycle were not increased. Acetate, glyoxylate, and malonate did not increase the specific nitrifying activity, but they increased the enzyme activities. These results imply that the involvement of acetate metabolism in the heterotrophic nitrification is unlikely.

Acetates↗

PubMed2004

Production of a novel polygalacturonic acid bioflocculant REA-11 by Corynebacterium glutamicum.

The production of a novel polygalacturonic acid bioflocculant REA-11 from a newly isolated strain, Corynebacterium glutamicum CCTCC M201005, was investigated. Sucrose was chosen as a carbon source for REA-11 production. Complex nitrogen sources containing urea and an organic nitrogen compound enhanced both bacterial growth and REA-11 production, among which urea plus corn steep liquor was shown to be the most efficient combination. A cost-effective medium for REA-11 production mainly comprised 17 g/l sucrose, 0.45 g/l urea, and 5 ml/l corn steep liquor, under which conditions the flocculating activity reached 390 U/ml. The molar ratio of carbon to nitrogen (C/N) significantly affected REA-11 production, where a C/N ratio of 20:1 was shown to be the best. Interestingly, by simultaneously feeding sucrose and urea at a C/N ratio of 20:1 at 24 h of fermentation, REA-11 production (458 U/ml) was enhanced by 17% compared to the control. In a 10 l jar fermentor, lower dissolved oxygen tension was favorable for REA-11 production: a flocculating activity of 520 U/ml was achieved at a kappaLa of 100 h(-1). REA-11 raw product is relatively thermo-stable at acidic pH ranges of 3.0-6.5. Preliminary application studies showed that REA-11 had stronger flocculating activity to Kaolin clay suspension compared to chemical flocculants. In addition, the capability of decolorizing molasses wastewater indicates the industrial potential of this novel bioflocculant.

Carbon↗

PubMed2005

In vitro bioactivity of combustion products from 12 tobacco constituents.

Twelve chemical components of tobacco leaf, representing 50% of its dry weight, were individually combusted and the bioactivities of their combustion products i.e. total particulate matter (TPM) were assayed using three in vitro tests. These components included carbohydrates, amino acids, proteins, polyphenols and carboxylic acids. The mutagenic potencies were assessed with the Salmonella mutagenicity assay (S. typhimurium TA98 and TA100). The induction of chromosomal damage, determined with the micronucleus test (IVMNT), and the neutral red uptake cytotoxicity test (NRU), were conducted on V79 hamster lung fibroblast cells. The Salmonella mutagenicity test and IVMNT were conducted with and without rat liver microsomal S9 fraction. Salmonella mutagenicity data confirmed the mutagenicity of TPM samples obtained from nitrogenous compounds (amino acids and proteins). The IVMNT showed that precursors of phenols in smoke (i.e. polyphenols) exhibited significantly higher levels of toxicity compared to other tobacco components. While S9 activation amplified the Salmonella mutagenicity response to combustion products, it significantly inhibited the toxicity measured with the IVMNT. NRU data demonstrated the increasing cytotoxicity induced following longer exposure time to TPM samples from nitrogenous and phenolic components. This study is the first to characterize the toxicity of the combustion products of major tobacco constituents. Our data suggest different mechanisms of toxicity and underline the relevance of using various bioassays.

Analysis of Variance↗

PubMed2003

Simultaneous nitrification and denitrification by controlling vertical and horizontal microenvironment in a membrane-aerated biofilm reactor.

Nitrogen and carbon components in domestic modified wastewater were completely removed by simultaneous nitrification and denitrification using a membrane-aerated biofilm reactor where biofilm was fixed on a hollow-fiber membrane. To measure the spatial distribution of pH, ammonium and nitrate ions and to observe microbes inside the biofilm fixed on the membrane, microelectrodes and the fluorescence in situ hybridization (FISH) method were applied. Due to plug flow in the vertical direction (from the bottom to the top of the reactor), ammonium nitrogen was gradually removed and negligible nitrate nitrogen was detected throughout the reactor. FISH revealed that ammonia-oxidizing bacteria were mainly distributed inside the biofilm and other bacteria, which included denitrifying bacteria, were mainly distributed outside the biofilm and over the suspended sludge. In order to characterize bacterial activity in the vertical direction of the reactor, nitrification rates at lower, central and upper points were calculated using microelectrode data. The nitrification rate at the lower point was 7 and 125 times higher than those at the central and upper points, respectively. These results show that the removal of carbon and nitrogen compounds was accomplished efficiently by using various kinds of bacteria distributed vertically and horizontally in a single reactor.

Bacteria, Aerobic↗

PubMed1980

Nitrogen catabolite repression of asparaginase II in Saccharomyces cerevisiae.

The biosynthesis of asparaginase II in Saccharomyces cerevisiae is subject to strong catabolite repression by a variety of nitrogen compounds. In the present study, asparaginase II synthesis was examined in a wild-type yeast strain and in strains carrying gdhA, gdhCR, or gdhCS mutations. The following effects were observed: (i) In the wild-type strain, the biosynthesis of asparaginase II was strongly repressed when either 10 mM ammonium sulfate or various amino acids (10 mM) served as the source of nitrogen. (ii) In a yeast strain carrying the gdhA mutation, asparaginase II was synthesized at fully derepressed levels when 10 mM ammonium sulfate was the source of nitrogen. When amino acids (10 mM) served as the nitrogen source, asparaginase II synthesis was strongly repressed. (iii) In a strain carrying the gdhCR mutation, the synthesis of asparaginase II was partially (30 to 40%) derepressed when either 10 mM ammonium sulfate or amino acids were present in the medium. (iv) In a yeast strain containing both gdhA and gdhCR mutations, asparaginase II synthesis was fully derepressed when 10 mM ammonium sulfate was the nitrogen source and partially derepressed when 10 mM amino acids were present. (v) Yeast strains carrying the gdhCS mutation were indistinguishable from the wild-type strain with respect to asparaginase II synthesis.

Amino Acids↗

PubMed1983

Effect of microbiological hydrolysis of urea on the nitrification process.

The aim of this study was to determine the causes of previously observed inhibition of stage II of nitrification in nitrifying activated sludge treating industrial wastewaters containing ammonium, urea and nitrates. Experiments made with both industrial wastewaters and synthetic medium containing equivalent concentrations of nitrogen compounds demonstrated that the factor limiting the activity of nitrobacteria is microbiological hydrolysis of urea, the causative agent probably being free ammonia released in the course of this hydrolysis.

Bacteria↗

PubMedDate not supplied

Carbon and nitrogen metabolism in ectomycorrhizal fungi and ectomycorrhizas.

The literature concerning the metabolism of carbon and nitrogen compounds in ectomycorrhizal associations of trees is reviewed. The absorption and translocation of mineral ions by the mycelia require an energy source and a reductant which are both supplied by respiratory catabolism of carbohydrates produced by the host plant. Photosynthates are also required to generate the carbon skeletons for amino acid and carbohydrate syntheses during the growth of the mycelia. Competition for photosynthates occurs between the fungal cells and the various vegetative sinks in the host tree. The nature of carbon compounds involved in these processes, their routes of metabolism, the mechanisms of control and the partitioning of metabolites between the various sites of utilization are only poorly understood. Both ascomycetous and basidiomycetous ectomycorrhizal fungi synthesize and some, if not all, accumulate mannitol, trehalose and triglycerides. The fungal strains employ the Embden--Meyerhof pathway of glucose catabolism and the key enzymes of the pentose phosphate pathway (6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase, transaldolase and transketolase). Anaplerotic CO2 fixation, via pyruvate carboxylase and/or phosphoenolpyruvate carboxykinase, provides high pools of amino acids. This process could be important in the recapture and assimilation of respired CO2 in the rhizosphere. The ectomycorrhizas are thought to contain the Embden--Meyerhof pathway, the pentose phosphate pathway and the tricarboxylic acid cycle, which provide the carbon skeletons for the assimilation of ammonia into amino acids. The main route of assimilation of ammonia appears to be through the glutamine synthetase-glutamate synthase cycle in the ectomycorrhizas. Glutamate dehydrogenase plays a minor role in this process. Glutamate dehydrogenase and glutamine synthetase are present in free-living ectomycorrhizal fungi and they participate in the assimilation of ammonia and the synthesis of amino acids through the glutamate dehydrogenase/glutamine synthetase sequence. In both in vitro cultures of fungi and ectomycorrhizas, the assimilated nitrogen accumulates in glutamine. Glutamine, but also ammonia, are thought to be exported from the fungal tissues to the host cells. Studies on the metabolism of ectomycorrhizas and ectomycorrhizal fungi have focused on the metabolic pathways and compounds which accumulate in the symbiotic tissues. Studies on regulation of the overall process, and the control of enzyme activity in particular, are still fragmentary.(ABSTRACT TRUNCATED AT 400 WORDS)

Carbon↗

PubMed1990

Changes in some nutrients of fenugreek (Trigonella Foenum graecum L.) seeds during water boiling.

Fenugreek seeds were boiled in water for various lengths of time (5, 10 and 15 min). Changes in weight, volume, moisture content, total sugars, nitrogen compounds, minerals, phosphorus compounds, phytic acid, amino acids and the in vitro digestibility of the seeds as well as the total solids of the boiling water were determined. Data indicated that there was an increase in both weight and volume as well as the in vitro digestibility of fenugreek seeds especially after the first 5 minutes of boiling. On the other hand, a decrease in the content of total sugars, protein compounds, calcium, magnesium, phytic acid, phosphorus and amino acids was observed. The reduction was accompanied by a gradual increase in the total solids of boiling water.

Amino Acids↗

PubMed1997

Influence of supplementary fish oil and rumen-protected methionine on milk yield and composition in dairy cows.

The effects of a daily supplement of 300 ml fish oil and 20 g rumen-protected methionine, alone or in combination, were investigated in mid-lactation cows receiving a maize silage-based diet ad lib. Fish oil significantly decreased feed intake, increased milk yield, decreased protein and casein concentrations, and especially fat concentration (by 13.1 g/kg) and output. Fat concentration decreased more in primiparous than in multiparous cows. Methionine supplementation increased protein and casein concentrations and outputs. No significant interaction between oil and methionine supplementation was found on milk composition. Treatments did not modify live weight or body condition changes, or lactose and non-casein nitrogenous compounds in milk. Oil plus methionine supplementation made it possible to decrease milk fat content without changing protein content.

Animals↗

PubMed1959

Enzyme biosynthesis in Escherichia coli.

Escherichia coli B synthesized beta-galactosidase and an enzyme system for D-xylose when exposed to lactose and xylose respectively in nitrogen-free media. The amount of beta-galactosidase formed in the absence of external nitrogen depended upon the nature of the medium in which the cells had originally been grown. Half as much of this enzyme was synthesized without exogenous nitrogen by cells taken from a nitrogen-rich medium as was formed by cells under favorable conditions with an external supply of nitrogen. Escherichia coli B contained a pool of nitrogen compounds soluble in 80 per cent ethanol and made up of several ninhydrin-positive components. One of these was identified chromatographically as glycine using an authentic radioactive sample. Another substance behaved like serine on the chromatograms. The internal pool of amino acids and peptides was large enough to account for the beta-galactosidase synthesized by cells exposed to lactose in a medium free of nitrogen. Some degree of interaction of the syntheses of the beta-galactosidase and xylose enzyme systems was observed in nitrogen-free media. This interaction produced a greater effect on the formation of beta-galactosidase and was attributed to a limiting factor(s) in the internal nitrogenous pool or to a limiting intermediate in enzyme synthesis.

Enzymes↗

PubMed1998

A diet containing chickpeas and wheat offers less protection against colon tumors than a casein and wheat diet in dimethylhydrazine-treated rats.

We examined the influence of extruded chickpeas and wheat relative to casein and wheat in a dimethylhydrazine (DMH)-induced colon tumor study in male Sprague-Dawley rats. The three diets, based on a modified AIN76 rodent diet with fat present at 10 g/100 g dry matter (DM), were as follows: casein with wheat starch (Cas/S) as control, casein with wheat (Cas/W) and chickpeas with wheat (CP/W). All diets were fed from 5 wk of age throughout the 28-wk study. At 28 wk, there was a significantly lower incidence of large intestinal tumors in rats fed Cas/W relative to those fed CP/W ( 11 vs. 56%, chi-square test, P = 0.018). The colonic tumor burden (tumors/tumor-bearing animal) was not different in Cas/W-fed and CP/W-fed rats (1 vs. 1.7), but the tumor mass index was significantly lower in the former group (0.22 vs. 1.21, P = 0.026). Rats fed the CP/W diet had significantly lower plasma cholesterol concentration (P < 0.01) than rats fed the other two diets. The cecal contents of rats fed the CP/W diet had significantly greater relative weights (46%, P < 0.05) than those of the Cas/W-fed rats; this was associated with higher concentrations of all short-chain fatty acids. Fecal analyses showed significantly (P < 0.05) higher concentrations of total fat (54%), total steroids (83%) and secondary bile acids (179%) in the CP/W-fed rats relative those fed Cas/W. There were higher concentrations of nitrogen in the feces of CP/W rats relative to the Cas/W-fed rats (84%, P < 0.05), associated with greater fecal weights (67%, P < 0.05). Although wheat and its fibers have been shown to be protective against DMH-induced cancers in rats, this was not the case in this study in which chickpeas (45 g/100 g diet) provided the protein and were an important source of soluble fiber. Elevated fat, secondary bile acid concentrations and/or nitrogenous compounds could be responsible for the increased colon tumorigenesis seen and may reflect a legume effect.

Animal Nutritional Physiological Phenomena↗

PubMed2007

Differences in postmortem urea nitrogen, creatinine and uric acid levels between blood and pericardial fluid in acute death.

Previous studies showed significant differences in postmortem urea nitrogen (UN), creatinine (Cr) and uric acid (UA) levels in heart blood depending on the causes of death, including acute death. In addition, the levels in pericardial fluid approximated the clinical serum reference ranges, and their elevations may be assessed based on clinical criteria. The present study investigated difference between blood and pericardial levels of these markers. Medicolegal autopsy cases (n=556, within 48h postmortem) of the following causes of death were examined: injury (n=136), asphyxiation (n=50), drowning (n=39), fire fatalities (n=99), hyperthermia (n=11), hypothermia (n=8), poisoning (n=26), delayed traumatic death (n=44) and natural diseases (n=143). When serum UN, Cr and UA levels were compared with the pericardial levels, there was an equivalency for delayed traumatic death and chronic renal failure, although each level was markedly elevated. Parallel increases in serum and pericardial UA and/or Cr levels were also observed for hypothermia and gastrointestinal bleeding. However, in drowning cases, the left cardiac and pericardial UN levels were lower than the right cardiac and peripheral levels, suggesting the influence of water aspiration. Significant elevations in serum and pericardial Cr and UA levels with a higher serum/pericardial UA ratio for fatal methamphetamine intoxication suggest progressive skeletal muscle damage due to advanced hypoxia/acidosis. Similar findings were often observed for other acute and subacute deaths. These findings suggest that a comparison between blood and pericardial nitrogenous compounds would be useful for investigating the cause and process of death.

Adolescent↗

PubMed2005

Nutrient balance in a paddy field with a recycling irrigation system.

We studied nutrient balance in a paddy field that had a recycling irrigation system and evaluated the effect of the irrigation system on nutrient balance during the irrigation period, from April to August 2002. Chemical fertilizer was the main input of phosphorus; the soil absorbed about 56% of it. The amount of nitrogen supplied by the irrigation system was higher than in a representative paddy field, and the amount of nitrogen fertilizer used was decreased because the irrigation water was partly reused. About 20% of applied nitrogen was lost by denitrification. The net outflows of phosphorus and nitrogen were -0.37 and -3.98 kg ha(-1), respectively. These results indicate that our study paddy field performed well in removing phosphorus and nitrogen compounds from runoff water. A recycling irrigation system can be considered an effective way of reducing the amounts of water and fertilizer used and reducing the outflow nutrients.

Conservation of Natural Resources↗