Dur3 and nrt2 genes in the bloom-forming dinoflagellate Prorocentrum minimum: Transcriptional responses to available nitrogen sources

Chemosphere ◽  
2020 ◽  
Vol 241 ◽  
pp. 125083
Author(s):  
S.A. Pechkovskaya ◽  
N.A. Knyazev ◽  
O.V. Matantseva ◽  
A.K. Emelyanov ◽  
I.V. Telesh ◽  
...  
2015 ◽  
Vol 112 (45) ◽  
pp. E6243-E6252 ◽  
Author(s):  
Stephan Klähn ◽  
Christoph Schaal ◽  
Jens Georg ◽  
Desirée Baumgartner ◽  
Gernot Knippen ◽  
...  

Glutamine synthetase (GS), a key enzyme in biological nitrogen assimilation, is regulated in multiple ways in response to varying nitrogen sources and levels. Here we show a small regulatory RNA, NsiR4 (nitrogen stress-induced RNA 4), which plays an important role in the regulation of GS in cyanobacteria. NsiR4 expression in the unicellularSynechocystissp. PCC 6803 and in the filamentous, nitrogen-fixingAnabaenasp. PCC 7120 is stimulated through nitrogen limitation via NtcA, the global transcriptional regulator of genes involved in nitrogen metabolism. NsiR4 is widely conserved throughout the cyanobacterial phylum, suggesting a conserved function. In silico target prediction, transcriptome profiling on pulse overexpression, and site-directed mutagenesis experiments using a heterologous reporter system showed that NsiR4 interacts with the 5′UTR ofgifAmRNA, which encodes glutamine synthetase inactivating factor (IF)7. InSynechocystis, we observed an inverse relationship between the levels of NsiR4 and the accumulation of IF7 in vivo. This NsiR4-dependent modulation ofgifA(IF7) mRNA accumulation influenced the glutamine pool and thusNH4+assimilation via GS. As a second target, we identifiedssr1528, a hitherto uncharacterized nitrogen-regulated gene. Competition experiments between WT and an ΔnsiR4KO mutant showed that the lack of NsiR4 led to decreased acclimation capabilities ofSynechocystistoward oscillating nitrogen levels. These results suggest a role for NsiR4 in the regulation of nitrogen metabolism in cyanobacteria, especially for the adaptation to rapid changes in available nitrogen sources and concentrations. NsiR4 is, to our knowledge, the first identified bacterial sRNA regulating the primary assimilation of a macronutrient.


2009 ◽  
Vol 100 (13) ◽  
pp. 3358-3365 ◽  
Author(s):  
Xiang Zou ◽  
Hai-feng Hang ◽  
Ju Chu ◽  
Ying-ping Zhuang ◽  
Si-liang Zhang

2017 ◽  
Vol 83 (5) ◽  
Author(s):  
Lucie Crépin ◽  
Nhat My Truong ◽  
Audrey Bloem ◽  
Isabelle Sanchez ◽  
Sylvie Dequin ◽  
...  

ABSTRACT During fermentative growth in natural and industrial environments, Saccharomyces cerevisiae must redistribute the available nitrogen from multiple exogenous sources to amino acids in order to suitably fulfill anabolic requirements. To exhaustively explore the management of this complex resource, we developed an advanced strategy based on the reconciliation of data from a set of stable isotope tracer experiments with labeled nitrogen sources. Thus, quantifying the partitioning of the N compounds through the metabolism network during fermentation, we demonstrated that, contrary to the generally accepted view, only a limited fraction of most of the consumed amino acids is directly incorporated into proteins. Moreover, substantial catabolism of these molecules allows for efficient redistribution of nitrogen, supporting the operative de novo synthesis of proteinogenic amino acids. In contrast, catabolism of consumed amino acids plays a minor role in the formation of volatile compounds. Another important feature is that the α-keto acid precursors required for the de novo syntheses originate mainly from the catabolism of sugars, with a limited contribution from the anabolism of consumed amino acids. This work provides a comprehensive view of the intracellular fate of consumed nitrogen sources and the metabolic origin of proteinogenic amino acids, highlighting a strategy of distribution of metabolic fluxes implemented by yeast as a means of adapting to environments with changing and scarce nitrogen resources. IMPORTANCE A current challenge for the wine industry, in view of the extensive competition in the worldwide market, is to meet consumer expectations regarding the sensory profile of the product while ensuring an efficient fermentation process. Understanding the intracellular fate of the nitrogen sources available in grape juice is essential to the achievement of these objectives, since nitrogen utilization affects both the fermentative activity of yeasts and the formation of flavor compounds. However, little is known about how the metabolism operates when nitrogen is provided as a composite mixture, as in grape must. Here we quantitatively describe the distribution through the yeast metabolic network of the N moieties and C backbones of these nitrogen sources. Knowledge about the management of a complex resource, which is devoted to improvement of the use of the scarce N nutrient for growth, will be useful for better control of the fermentation process and the sensory quality of wines.


2014 ◽  
Vol 2 (1) ◽  
pp. 41-44 ◽  
Author(s):  
Jarina Joshi

Ammonium sulphate, ammonium phosphate, sodium nitrate, urea and glycine were the five different commonly available nitrogen sources used at different concentration ranging from 0.5 to 4% w/v to produce ethanol in batch culture. Potato paste made from red potatoes grown in hilly regions of Nepal was used as carbon source. Prior to fermentation all carbon sources were saccharified enzymatically using α- amylase at pH 5 and temperature 55oC. Maximum yield of ethanol 5.2% was obtained at a temperature of 30oC and pH 5.0 without exogeneous supply of nitrogen. There is slight decrease in concentration when temperature is decreased to 25oC but a drastic decrease in concentration when temperature is increased beyond optimum. All the exogeneously supplied nitrogen sources found to enhance ethanol production and cell viability when yeast strain Saccharomyces cerevisiae isolated from brewer’s yeast was used. Ammonium sulphate was found as best nitrogen supplement among them. Maximum ethanol percentage of 8.3 was observed at pH 5.0 and temperature 30oC with Ammonium sulphate concentration of 2%.DOI: http://dx.doi.org/10.3126/ijasbt.v2i1.9191Int J Appl Sci Biotechnol, Vol. 2(1): 41-44


2012 ◽  
Vol 2012 ◽  
pp. 1-7 ◽  
Author(s):  
Haytham M. El-Sharkawi

Knowledge to increase the microbial biomass nitrogen (MBN) as a bulk of free-living microbes in paddy soil is limited. The potential benefit of these microorganisms was evaluated, in this study, under different nitrogen sources and two paddy soils. The results revealed that pots treated with organic matter recorded the maximum value of the total N uptake and MBN, followed by the Urea treated pots. Pots amended with sludge exhibited a higher microbial N forming ability than those amended with straw compost under both soils. But ammonium concentration in soil increased with straw compost application. Under fresh soil treatment, microbial N uptake rate and proportion of plant nitrogen derived from microbial nitrogen sources () were higher than autoclaved soil. A positive correlation was found between the and the total N in rice shoot in both soils. Finally, we can say that MBN was governed not only by the soil nitrogen content but also by the type of the nitrogen source. The addition of sludge to fresh soil increased total MBN and consequently could be indirectly beneficial to rice production especially in poor soils. Thus, soil microbes contribute to plant growth by serving the available nitrogen during the season.


2012 ◽  
Vol 78 (19) ◽  
pp. 6838-6849 ◽  
Author(s):  
Vernita J. Reid ◽  
Louwrens W. Theron ◽  
Maret du Toit ◽  
Benoit Divol

ABSTRACTThe extracellular acid proteases of non-Saccharomyceswine yeasts may fulfill a number of roles in winemaking, which include increasing the available nitrogen sources for the growth of fermentative microbes, affecting the aroma profile of the wine, and potentially reducing protein haze formation. These proteases, however, remain poorly characterized, especially at genetic level. In this study, two extracellular aspartic protease-encoding genes were identified and sequenced, from two yeast species of enological origin: one gene fromMetschnikowia pulcherrimaIWBT Y1123, namedMpAPr1, and the other gene fromCandida apicolaIWBT Y1384, namedCaAPr1.In silicoanalysis of these two genes revealed a number of features peculiar to aspartic protease genes, and both the MpAPr1 and CaAPr1 putative proteins showed homology to proteases of yeast genera. Heterologous expression ofMpAPr1inSaccharomyces cerevisiaeYHUM272 confirmed that it encodes an aspartic protease. MpAPr1 production, which was shown to be constitutive, and secretion were confirmed in the presence of bovine serum albumin (BSA), casein, and grape juice proteins. TheMpAPr1gene was found to be present in 12 otherM. pulcherrimastrains; however, plate assays revealed that the intensity of protease activity was strain dependent and unrelated to the gene sequence.


2007 ◽  
Vol 7 (4) ◽  
pp. 604-620 ◽  
Author(s):  
Viktor M. Boer ◽  
Siew Leng Tai ◽  
Zeynep Vuralhan ◽  
Yalun Arifin ◽  
Michael C. Walsh ◽  
...  

2020 ◽  
Author(s):  
Yu Chen ◽  
Eunice van Pelt-KleinJan ◽  
Berdien van Olst ◽  
Sieze Douwenga ◽  
Sjef Boeren ◽  
...  

Cells adapt to different conditions via gene expression that tunes metabolism and stress resistance for maximal fitness. Constraints on cellular proteome may limit such expression strategies and introduce trade-offs1; Resource allocation under proteome constraints has emerged as a powerful paradigm to explain regulatory strategies in bacteria2. It is unclear, however, to what extent these constraints can predict evolutionary changes, especially for microorganisms that evolved under nutrient-rich conditions, i.e., multiple available nitrogen sources, such as the lactic acid bacterium Lactococcus lactis. Here we present an approach to identify preferred nutrients from integration of experimental data with a proteome-constrained genome-scale metabolic model of L. lactis (pcLactis), which explicitly accounts for gene expression processes and associated constraints. Using glucose-limited chemostat data3, we identified the uptake of glucose and arginine as dominant constraints, whose pathway proteins were indeed upregulated in evolved mutants. However, above a growth rate of 0.5 h-1, pcLactis suggests that available enzymes function at their maximum capacity, which allows an increase in growth rate only by altering gene expression to change metabolic fluxes, as was mainly observed for arginine metabolism. Thus, our integrative analysis of flux and proteomics data with a proteome-constrained model is able to identify and explain the constraints that form targets of regulation and fitness improvement in nutrient-rich growth environments.


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