Effect of Dipeptides on the Growth of <Emphasis Type="Italic">Oenococcus oeni</Emphasis> in Synthetic Medium Deprived of Amino Acids

Oenococcus oeni has numerous amino acid requirements for growth and dipeptides could be important for its nutrition. In this paper the individual or combined effect of dipeptides on growth of O. oeni X₂L in synthetic media deficient in one or more amino acids with L-malic acid was investigated. Utilization of dipeptides, glucose, and L-malic acid was also analyzed. Dipeptides were constituted by at least one essential amino acid for growth. Dipeptides containing two essential amino acids, except leucine, had a more favorable effect than free amino acids on the growth rate. Gly-Gly was consumed to a greater extent than Leu-Leu and a rapid exodus of glycine to the extracellular medium accompanied it. The microorganism could use glycine in exchange for other essential amino acids outside the cell, favoring growth. In the presence of Leu-Leu, the increase in glucose consumption rate could be related to the additional energy required for dipeptide uptake.     

Injection of essential amino acids substitutes for bacterial supply in aposymbiotic pea aphids (Acyrthosiphon pisum)

The symbiotic bacteria Buchnera contribute to the nutrition of pea aphids, Acyrthosiphon pisum, through the provision of essential amino acids which are lacking in the diet. However, chemically defined diets, containing nutritionally adequate amounts of essential amino acids, fail to rescue aposymbiotic aphids, in which the bacteria have been disrupted with antibiotics. In this study the injection of a mixture of essential amino acids into the haemocoel of aposymbiotic aphids was shown to alleviate, at least partially, the impact of symbiont loss. Specifically, the total amino acid content in the tissues of aposymbiotic aphids was reduced by approximately 40% to levels comparable with symbiotic insects, and there was a 1.7-fold increase in the number of embryos, suggesting that the availability of essential amino acids promotes aphid protein synthesis by rejuvenating the free amino acid pool of aposymbiotic aphids. In addition, a similar effect on the total amino acid content was observed when phenylalanine alone, but not glutamine, lysine or tryptophan, was injected into the haemocoel of aposymbiotic aphids, and there was also a significant increase in the number of embryos following injection of phenylalanine or tryptophan alone. The impact of amino acid injection on the embryo complement of aposymbiotic aphids was limited to an increase in the number of embryos, with no increase in basal embryo size. It is proposed that older embryos may rely on their own complement of symbiotic bacteria for essential amino acid provisioning. Taken together, the data highlight the importance of bacterial provisioning of essential amino acids, particularly the aromatic amino acids, in the intact symbiosis.     

The effect of cold shock on the free amino acid pool of rotan pondfish <Emphasis Type="Italic">Perccottus glehni</Emphasis> (<Emphasis Type="Italic">Eleotridae,Perciformes</Emphasis>)

The effect of cold shock on the free amino acid pool of the pondfish P. glehni not adapted to hibernation has been studied. In the beginning of September, after four days of cold shock, a decrease in the blood level of essential amino acids and an increase in the level of alanine and three-fold increase in the level of γ-aminobutyric acid (GABA) were observed. Changes in the free amino acid pool in the mussels occur in two stages. Namely, in a considerable decrease in their concentration after two days of cold shock followed by an even more significant increase in their amount after four days of shock. In the brain changes in the free amino acid pools are associated with their total decrease followed by continuous significant decrease in the level of taurine during the first four days of cold shocking. The obtained results are compared with previously obtained data on the free amino acid content in muscles and blood of P. glehni during preparation for winter sleep and the beginning of hibernation.     

Growth Response and Modifications of Organic Nitrogen Compounds in Pure and Mixed Cultures of Lactic Acid Bacteria from Wine

The interactions between the proteolytic X₂L strain of Oenococcus oeni and the non-proteolytic 12p strain of Pediococcus pentosaceus were assayed. The characteristics of cell growth, protein degradation, and amino acid production of both strains were determined in pure and mixed cultures. O. oeni showed poor cell growth and greater ability in the release of amino acids to the extracellular medium, whereas P. pentosaceus showed a higher yield in cell production with a decrease in the amino acid concentration in the medium. P. pentosaceus especially consumed essential amino acids for growth, and O. oeni released several of the essential amino acids important for growth of P. pentosaceus. In the mixed culture, mutualism was observed. The higher activity of the proteolytic system of O. oeni in mixed culture produced an increase in cell growth and in the amount of essential amino acids released. These findings provide new knowledge about the metabolic interactions between lactic acid bacteria isolated from wine when proteins are degraded in mixed bacterial populations.     

Protein increase and lysine production by a <Emphasis Type="Italic">Paecilomyces variotii</Emphasis> strain grown on two-phase olive mill waste

Two-phase olive-mill waste, the so-called “ecological”, has been treated with a Paecilomyces variotii isolate in solid state fermentation experiments. The growth of the microorganism was estimated by measuring the production of carbon dioxide, using gas chromatography. A 46% increase of the protein content was achieved at the fermented product, after molasses addition at the initial mixture. The amino acid profile of the produced protein, as far as the essential amino acids are concerned, was significantly improved, resulting in a product that has the potential to be used as animal feed. Furthermore, it contains lysine, one of the essential amino acids that did not exist at the original product and is produced during fermentation. This is the first report on solid state fermentation of the two-phase olive mill waste (TPOMW) as a substrate, using a Paecilomyces variotii strain.     

Studies on the influence of foliar nutrient sprays on the root exudation pattern in four crop plants

Summary Studies conducted to examine the exudation pattern of amino acids and sugars in four crop plants,viz sorghum, sunnhemp, ragi, and tomato indicated that in all, 17 known amino acids and 4 sugars were exuded and that the number and nature of the exuded amino acids and sugars differed with the plant species and with the age of plant. Glutamic and aspartic acids were found to be present in the exudates of all the plant species at all stages of plant growth examined. The quantities of amino acids and sugars differed with plant species and the maximum quantity of the chemicals was exuded during the early stages of plant growth. Glutamic acid among amino acids, and glucose among sugars, were always present in higher concentrations than the others, in the exudates in all the four crop plants.Foliar application of nitrogen in the form of NaNO₃ and phosphorus as Na₂HPO₄, was found to alter the exudation pattern of amino acids and sugars and such influence differed in different plant species. There was a general increase in the total concentration of amino acids and a decrease in sugar content in the exudates after treatment of the foliage with N, while a decrease in the amino acid content and increase in total sugars with P-treatment was observed.     

Functional analysis of the cysteine motifs in the ferredoxin-like protein FdxN of Rhizobium meliloti involved in symbiotic nitrogen fixation.

Summary TheRhizobium meliloti fdxN gene, which is part of thenifA-nifB fdxN operon, is absolutely required for symbiotic nitrogen fixation. The deduced sequence of the FdxN protein is characterized by two cysteine motifs typical of bacterial-type ferredoxins. The Fix^– phenotype of anR. meliloti fdxN: :[Tc] mutant could be rescued by theR. leguminosarum fdxN gene, whereas no complementation was observed withnif-associated genes encoding ferredoxins fromBradyrhizobium japonicum, Azotobacter vinelandii, A. chroococcum andRhodobacter capsulatus. In addition to these heterologous genes, severalR. meliloti fdxN mutant genes constructed by site-directed mutagenesis were analyzed. Not only a cysteine residue within the second cysteine motif (position 42), which is known to coordinate the Fe-S cluster in homologous proteins, but also a cysteine located down-stream of this motif (position 61), was found to be essential for the activity of theR. meliloti FdxN protein. Changing the amino acid residue proline in position 56 into methionine resulted in a FdxN mutant protein with decreased activity, whereas changes in positions 35 (Asp35Glu) and 45 (Gly45Glu) had no significant effect on the function of the FdxN mutant proteins. In contrast to bacterial-type ferredoxins, which contain two identical cysteine motifs of the form C-X₂-C-X₂-C-X₃-C,nif-associated ferredoxins, includingR. meliloti FdxN, are characterized by two different cysteine motifs. Six additional amino acids separate the second (Cys₄₂) and the third cysteine (Cys₅₁) in the C-terminal motif (C-X₂-C-X₈-C-X₃-C). By molecular modelling, it was predicted that these amino acid residues form a loop, which does not alter the relative positions of the neighbouring cysteines. Deletion of this loop resulted in anR. meliloti FdxN mutant protein, which exhibited almost 70% wild-type activity, indicating that the predicted loop does not affect Fe-S cluster binding and plays no crucial role in activity of the FdxN protein.     

Über die Wirkung kurzfristiger Kinetingaben auf Protein,Aminosäuren und RNS bei Lemna minor L.

Summary Lemna minor L. was cultivated on nutrient medium with kinetin (10^-5 m) for 3 hours at 28°C and ca. 5,000 lux. Samples were taken at different times and their amino acid, protein and RNA content was determined.The amino acid content in the cultures with kinetin corresponds to that in the cultures without kinetin. The amino acid concentration seems to be regulated by mechanisms independent of kinetin. On the other hand, kinetin increases the quantity of protein immediately i.e., within the first two minutes. This increase stops after ca. 30 minutes, but the excess of protein remains for a long time. In the first 30 minutes the RNA concentration also rises. This increase is not as strong as that of the protein nor does it remain. After 30 minutes, when the rise of protein content stops, the RNA concentration begins to drops within 1 hour to that of the control cultures without kinetin.The extent of the protein and RNA increase induced by kinetin depends strongly upon the physiological state of the plants. This may be because the plants produce different quantities of endogenous cytokinins under different conditions.The protein induced by kinetin was then examined after centrifugation at 100,000 x g. In samples taken during the first 5 minutes, the quantity of protein in the supernatant and in the sediment is increased. In samples taken after this period, the increase of protein in the supernatant disappears and only that in the sediment remains. After some time, therefore, all the excess protein consists of structural protein, which is found in the sediment. The findings are compared with those of other authors and an unspecific stimulation of RNA is suggested.     

Effect of insulin on folic-acid transport in cultured fibroblasts

Uptake of folic acid was measured in secondary cultures of skin fibroblasts from fetal rats. The cultures were made quiescent by 24 hours preincubation in medium containing 1% serum and subsequent 3 hours preincubation in phosphate buffered saline. The uptake of ³H-folic acid was linear with time during 15 seconds and reached a plateau level at 2–3 minutes. There was no further increase in the intracellular radioactivity until the end of the experiments at 10 minutes. The uptake of folic acid in fibroblasts was not concentrative and proceeded until equilibration with the extracellular concentration. Intracellular metabolic conversion of folic acid was not significant during the time of the experiments (up to 10 minutes). Insulin caused a two-fold increase in the initial rate of folate uptake as determined from the 15 second uptake values. The dose response curves for the insulin effect showed that 85% of the maximal effect was exerted by 1 m?M insulin. A lag period of 7–10 minutes was observed after the addition of insulin and before the effect on folic acid uptake was manifested. Thereafter the effect increased with the time of preincubation with insulin. The concentration dependence of folate uptake yielded non homogeneous curves. At low concentrations of substrate, saturable components were observed while at high concentration (above 5 × 10^?6 M) a linear component was observed. Insulin increased the slope of the linear component and the V_max of the saturable component while the K_m remained unaltered.     

Changes in intracellular amino acids and organic acids induced by nitrogen starvation and nitrate or ammonium resupply in the cyanobacterium Phormidium laminosum

In Phormidium laminosum cells, nitrogen starvation caused a decrease in the intracellular levels of all amino acids, except glutamate, and an increase in the total level of the analyzed organic acids. The addition of nitrate or ammonium to N-starved cells resulted in substantial increases in the pool size of most amino acids. Upon addition of ammonium the total level of organic acids diminished, whereas it increased upon addition of nitrate, after a transient decay during the first minutes. Nitrogen resupply stimulated amino acid synthesis, the effect being faster and higher when ammonium was assimilated. The data indicate that nitrate and ammonium assimilation induced an enhancement of carbon flow through the glycolytic and the tricarboxylic-acid pathways to amino acid biosynthesis, with a concurrent decrease in the carbohydrate reserves. The results suggest that the availability of carbon skeletons limited the rate of ammonium assimilation, whereas the availability of reducing equivalents limited the rate of nitrate assimilation.Abbreviations Chl chlorophyll - GOGAT ferredoxin-dependent glutamate synthase (EC 1.4.7.1) - GS glutamine synthetase (EC 6.3.1.2) This work has been supported by grants from the Spanish Ministry of Education and Science (DGICYT and PB92-0464) and the University of the Basque Country (042.310-EC203/94) M.I.T. and J.A.G. were the recipients of fellowships from the Basque Government.     

Effect of some pre-treatments on UV-sensitivity and on the course of macromolecular synthesis inEscherichia coli 15 T−, U−, his−

The UV-sensitivity ofEscherichia coli 15 T⁻, U⁻, his⁻ cells after a 45 minutes glucose, thymine uracil, or histidine pre-irradiation starvation, as well as the course of DNA, RNA, and protein synthesis during starvation and during a 60 minute post-treatment in a complete medium were investigated. An increased radioresistance was observed when starvation for some compounds resulted in a consequent inhibition of protein synthesis, as it was observed in the case of glucose, histidine, or uracil starvation. During thymine starvation, which led to a decreased resistance, no inhibition of protein synthesis was recorded. The postirradiation time-course of DNA synthesis did not show any correlation with the increased rate of resistance. The DNA synthesis after U⁻ pre-treatment was greatly delayed, however, after glucose pre-treatment no retardation was observed although both factors increased the rate of surviving cells approximately to the same extent. We assume that the factors which increase the radio-resistance could act by a similar mechanism which would take part in the inhibition of protein synthesis. This mechanism could consist in a decrease of the m-RNA turnover.     

Influence of stringent and relaxed response on excretion of recombinant proteins and fatty acid composition in Escherichia coli

In contrast to stringent (relA⁺) cells of Escherichia coli, relaxed (relA) cells excreted recombinant proteins (-lactamase, interferon 1) into the culture medium during amino acid limitation. Comparative analyses of overall fatty acid composition in relA⁺ cells and relA cells were performed and revealed that, in wild-type cells, drastic alterations occurred during the stringent response. The portion of saturated fatty acids (C16:0) and the fractions of cyclopropane fatty acids (C17_cyc and C19_cyc) increased whereas the portions of unsaturated fatty acids (C16:1 and C18:1) decreased. In cells of the relaxed mutant, no significant changes in the overall composition of the fatty acids were observed after the onset amino acid limitation. These results indicate that a change in fatty acid composition of membrane lipids after starvation of cells may be responsible for the prevention of loss of cellular proteins into the culture medium in stringent controlled cells of Escherichia coli.     

Effects of Anoxia on the Stimulated Release of Amino Acid Neurotransmitters in the Cerebellum In Vitro

Abstract: The effect of anoxia and ischemia on the release of amino acid transmitters from cerebellar slices induced by veratridine or high [K⁺] was studied. Synaptic specificity was tested by examining the tetradotoxin (TTX)-sensitive and the Ca²⁺-dependent components of stimulated release. Evoked release of endogenous amino acids was investigated in addition to more detailed studies on the stimulated efflux of preloaded [¹⁴C]GABA and d -[³H]aspartate (a metabolically more stable anologue of acidic amino acids).[¹⁴C]GABA release evoked by either method of stimulation was unaffected by periods of up to 35 min of anoxia and declined moderately by 45 min. In contrast, induced release of d -[³H]Asp increased markedly during anoxia to a peak at about 25 min, followed by a decline when anoxia was prolonged to 45 min. Evidence was obtained that the increased evoked efflux of d -[³H]Asp from anoxic slices was not due to impaired reuptake of the released amino acid and that it was completely reversible by reoxygenation of the slices. Results of experiments examining the evoked release of endogenous amino acids in anoxia were consistent with those obtained with the exogenous amino acids. Only 4 of the 10 endogenous amino acids studied exhibited TTX-sensitive veratridine-induced release under aerobic conditions (glutamate, aspartate, GABA, and glycine). Anoxia for 25 min did not affect the stimulated efflux of these amino acids with the exception of glutamate, which showed a significant increase. Compared with anoxia, effects of ischemia on synaptic function appeared to be more severe. Veratridine-evoked release of [¹⁴C]GABA was already depressed by 10 min and that of d -[³H]Asp showed a modest elevation only at 5 min. Stimulated release of d -Asp and labelled GABA declined progressively after 5 min. These findings were compared with changes in tissue ATP concentrations and histology. The latter studies indicated that in anoxia the earliest alterations are detectable in glia and that nerve terminals were the structures by far the most resistant to anoxic damage. The results thus indicated that evoked release of amino acid transmitters in the cerebellum is compromised only by prolonged anoxia in vitro. In addition, it would appear that the stimulated release of glutamate is selectively accentuated during anoxia. This effect may have a bearing on some hypoxic behavioral changes and, perhaps, also on the well-known selective vulnerability of certain neurons during hypoxia.     

Phloem amino acids and the host plant range of the polyphagous aphid, Aphis fabae

This study investigated the relationship between the essential amino acid requirement of the aphid Aphis fabae Scop. and the phloem sap amino acid composition of its host plants. The dietary amino acid requirement of A. fabae varied between clones. One or more of the eight clones of A. fabae tested displayed depressed larval survival, larval growth rate, or r_m on diets lacking histidine, methionine, threonine, and valine, but none of the other five essential amino acids. The required amino acids corresponded closely to the essential amino acids that varied in relative concentrations among 16 plant species tested: histidine, threonine, tryptophan, and valine. It is suggested that the interclonal variation in the dietary requirements of an aphid species may contribute to the intraspecific variation in plant utilisation patterns. The phloem sap amino acid composition and sucrose : amino acid ratio did not differ consistently between host plant species of A. fabae and non‐host species, indicating that phloem amino acid composition is not an important factor in determining the host plant range of this aphid species.     

Short- and Long-term Effects of High Temperatures (47–49 °C) on Tobacco Leaves. I. Photosynthesis

Detached Nicotiana rustica leaves were exposed for 2 minutes to temperatures in the range 36° to 51°C. Above 45°C, ¹⁴CO₂ fixation was reduced by half as compared with controls. The fall in ¹⁴CO₂ fixation continued for 2 1/2 h. Recovery was completed 45 h after the treatment. Above 45°C there was an increase in the labeled cationic and anionic fractions and a decrease in the neutral fractions, both increase and decrease being associated with the impaired CO₂ fixation. However, these changes in products were also demonstrated when leaves were exposed to the high temperature after labeling.     

不同酸度酸雨对小麦花后氮硫代谢和籽粒蛋白组分的影响

酸雨是中国重要的环境问题,为研究酸雨对小麦籽粒品质的可能影响,以小麦品种扬麦15和汶农17为材料开展盆栽试验,研究了不同酸度(pH2.5、p H4.0和p H5.6)酸雨对小麦花后氮硫代谢关键酶活性和籽粒蛋白质含量及组分的影响。结果显示:酸雨处理抑制叶片硝酸还原酶(NR)活性,提高了扬麦15整个灌浆期及汶农17灌浆中后期叶片谷氨酰胺合成酶(GS)活性,促进了叶片蛋白的降解,降低了叶片可溶性蛋白含量。不同酸度酸雨提高了成熟期籽粒中蛋白质含量,酸度越强,增加幅度越大,籽粒中各蛋白组分含量和大部分氨基酸含量也有明显提高。酸雨提高了扬麦15叶片丝氨酸乙酰转移酶(SAT)和O-乙酰丝氨酸硫裂解酶(OAS-TL)活性,但对汶农17硫代谢关键酶活性影响较小,酸雨处理还提高了籽粒中二硫键和含硫氨基酸含量。可见酸雨对小麦氮硫代谢有不同程度影响,进而影响了小麦籽粒蛋白质含量和组成,酸度越强影响越大,但不同品种对酸雨响应有一定差异。     

RNA removal from Candida utilis by chemical and thermal treatments

The removal of the content of nucleic acids of fodder yeast (Candida utilis) by treatment with HCl or heat shock was investigated Acid concentrations between 5 and 15% (on dried matter basis) were used. A maximal removal of the content of nucleic acids of 88% was realized wheńn was worked with 15% HCl, 90 °C during 30 minutes. But under this conditions were observed high demanges of protein and also of some essential amino acids Good results for diminishing the content of nucleic acids were reached with the highest concentration of acid and a treatment during 20 minutes The experiments with heat shock were carried out at 68 °C, different times for heating and different contents of yeast In this way better results than for treatment with acid according to diminishing the content of nucleic acids and yield of protein and essential amino acids were reached A removal of over 80% of the content of nucleic acids was achieved in all cases.     

Role of carbon dioxide fixation,blood aspartate and glutamate in the adaptation of amphibian brain tissues to a hyperosmotic internal environment

Mechanisms have been examined by which hyperosmotic blood plasma might elevate the levels of aspartate and glutamate in the brain of the toadBufo boreas. CO₂ fixation was assessed by two in vivo methods using [2-¹⁴C]glucose injected intracisternally. Thirty minutes after injection, the¹⁴C labeling of glutamate and aspartate was more than 100 times greater in brain than in liver. In brain tissues, 40+% of¹⁴C atoms appeared to be incorporated into aspartate via the pyruvate carboxylase pathway. Brain tissues of control toads and toads adapting or adapted to hyperosmotic plasma osmolality revealed no differences in the rate of CO₂ fixation as related to glucose utilization or tissue pool sizes of glutamate and aspartate. Elevated levels of these amino acids in blood plasma preceded increases in brain tissues. Carbon atoms required during hyperosmotic adaptation for expansion of amino acid pools in brain tissues may, in part, originate from amino acids in blood but apparently not from CO₂ fixation in brain.     

Seasonal patterns of nucleic acid concentrations and amino acid profiles of Parapenaeus longirostris (Crustacea, Decapoda): relation to growth and nutritional condition

The present work describes the seasonal changes in nucleic acid concentrations and amino acid profiles in the muscle of juvenile Parapenaeus longirostris and their relation to growth and nutritional condition. RNA content varied significantly between seasons, being the highest values attained in spring and the lowest in winter (p < 0.05). Similar results were obtained with RNA:protein and RNA:DNA ratios. In respect to total amino acid content (TAA), a significant increase from winter to spring was observed (p < 0.05) and the major essential amino acids (EAA) were arginine, histidine and leucine. Within non-essential amino acids (NEAA) glutamic acid, aspartic acid, glycine and proline were dominant. From winter to spring, a significant variation in NEAA content occurred (26.8; p < 0.05), mainly due to the significant increase of glutamic acid (79.1) and serine (66.7) (p < 0.05). EAA content did not vary significantly between seasons (p > 0.05). In opposition, during this period a significant decrease in the free amino acid content (FAA) was observed (p < 0.05); a higher percentage of decrease was attained in free non-essential (FNEAA – 42.9) in comparison to free essential amino acids (FEAA – 40.2). The significant increase in RNA and TAA contents from winter to spring may be related with protein synthesis. On the other hand, the lowest values obtained in winter may be due to a reduction in feeding activity; in this period the muscle protein must be progressively hydrolysed, which is evident with the higher FAA content. The liberated amino acids enter FAA pool and become available for energy production. In conclusion, it was evident that the seasonal cycle in activities such as feeding and growth with nucleic acids and amino acid analyses was noticed.     

Changes in phosphometabolites and intracellular pH in the tail muscle of the prawnPalaemon serratus as shown by in vivo31P-NMR

Summary ³¹P NMR spectra were recorded from tail muscles of the prawnPalaemon serratus, at rest, after exhaustive work and during subsequent recovery. At rest, the spectra indicated concentrations of phosphoarginine and ATP in good agreement with those obtained from resting fast skeletal muscles in mammals, which are characterized by a high phosphocreatine/P_i ratio. Following exhaustive work, phosphoarginine dropped by ca. 60% and ATP by 20%, while inorganic phosphate increased by 160%. The increase in inorganic phosphate immediately after contractions and in the first minutes of recovery corresponded partially to the changes in phosphoarginine and ATP. During recovery, the decrease of inorganic phosphate balanced the resynthesized phosphoarginine which was fully replenished within 30–40 min. The position of the inorganic phosphate resonance peak was used to monitor changes in intracellular pH (pH_i). The average pH_i in resting tail muscles was 7.20. After stimulation it was observed to decrease by 0.22 units. The return to pre-stimulation value was not achieved within 45 min. A NMR index (ATP+Arg-P)/(ATP+Arg-P+P_i) was calculated to characterize the extent of energetic changes caused by exercise.