Nitrogen metabolism and chloramphenicol production in Streptomyces venezuelae

1983 ◽  
Vol 29 (12) ◽  
pp. 1706-1714 ◽  
Author(s):  
S. Shapiro ◽  
L. C. Vining
Keyword(s):  
Growth Rate ◽  
Glutamate Dehydrogenase ◽  
Nitrogen Metabolism ◽  
Nitrogen Assimilation ◽  
Glutamate Synthase ◽  
Biomass Accumulation ◽  
Nitrogen Sources ◽  
Ph Control ◽  
Ammonium Nitrogen ◽  
Streptomyces Venezuelae

The relationship between chloramphenicol production and nitrogen metabolism in Streptomyces venezuelae was examined in stirred jar cultures under pH control. Nitrogen sources that supported rapid biomass accumulation gave low rates of antibiotic synthesis during growth. This was consistent with a general incompatibility between fast growth and high yields of chloramphenicol. In media where the growth rate was reduced below the attainable maximum by the rate at which nitrogen could be assimilated, chloramphenicol production was associated with biomass accumulation. Enzymes that are potentially associated with nitrogen assimilation pathways were assayed in cultures supplied with nitrogen sources supporting markedly different growth rates. The results indicated mat glutamine synthetase and alanine dehydrogenase levels were relatively insensitive to changes in growth rate and nitrogen source depletion. Glutamate dehydrogenase and glutamate synthase, on the other hand, showed high activity in cultures assimilating ammonium nitrogen and markedly decreased activity with poorer nitrogen sources or when ammonium was depleted. If chloramphenicol biosynthesis is coordinately controlled by mechanisms that regulate nitrogen assimilation, glutamate synthase and glutamate dehydrogenase are the most likely enzymes that manifest the regulatory linkage.

scholarly journals Expanded Role for the Nitrogen Assimilation Control Protein in the Response of Klebsiella pneumoniae to Nitrogen Stress

2010 ◽  
Vol 192 (19) ◽  
pp. 4812-4820 ◽  
Author(s):  
Ryan L. Frisch ◽  
Robert A. Bender
Keyword(s):  
Growth Rate ◽  
Stress Response ◽  
Klebsiella Pneumoniae ◽  
Nitrogen Metabolism ◽  
Nitrogen Limitation ◽  
Nitrogen Assimilation ◽  
Nitrogen Sources ◽  
Nitrogen Stress ◽  
A Genome ◽  
Regulated Promoters

ABSTRACT Klebsiella pneumoniae is able to utilize many nitrogen sources, and the utilization of some of these nitrogen sources is dependent on the nitrogen assimilation control (NAC) protein. Seven NAC-regulated promoters have been characterized in K. pneumoniae, and nine NAC-regulated promoters have been found by microarray analysis in Escherichia coli. So far, all characterized NAC-regulated promoters have been directly related to nitrogen metabolism. We have used a genome-wide analysis of NAC binding under nitrogen limitation to identify the regions of the chromosome associated with NAC in K. pneumoniae. We found NAC associated with 99 unique regions of the chromosome under nitrogen limitation. In vitro, 84 of the 99 regions associate strongly enough with purified NAC to produce a shifted band by electrophoretic mobility shift assay. Primer extension analysis of the mRNA from genes associated with 17 of the fragments demonstrated that at least one gene associated with each fragment was NAC regulated under nitrogen limitation. The large size of the NAC regulon in K. pneumoniae indicates that NAC plays a larger role in the nitrogen stress response than it does in E. coli. Although a majority of the genes with identifiable functions that associated with NAC under nitrogen limitation are involved in nitrogen metabolism, smaller subsets are associated with carbon and energy acquisition (18 genes), and growth rate control (10 genes). This suggests an expanded role for NAC regulation during the nitrogen stress response, where NAC not only regulates genes involved in nitrogen metabolism but also regulates genes involved in balancing carbon and nitrogen pools and growth rate.

Glutamate synthase levels in Neurospora crassa mutants altered with respect to nitrogen metabolism

1981 ◽  
Vol 1 (2) ◽  
pp. 158-164
Author(s):  
N S Dunn-Coleman ◽  
E A Robey ◽  
A B Tomsett ◽  
R H Garrett
Keyword(s):  
Neurospora Crassa ◽  
Glutamate Dehydrogenase ◽  
Nitrogen Metabolism ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Glutamate Synthase ◽  
Wild Type ◽  
Mutant Strains ◽  
Synthase Regulation ◽  
Glutamate Biosynthesis ◽  
Glutamine Limitation

Glutamate synthase catalyzes glutamate formation from 2-oxoglutarate plus glutamine and plays an essential role when glutamate biosynthesis by glutamate dehydrogenase is not possible. Glutamate synthase activity has been determined in a number of Neurospora crassa mutant strains with various defects in nitrogen metabolism. Of particular interest were two mutants phenotypically mute except in an am (biosynthetic nicotinamide adenine dinucleotide phosphate-glutamate dehydrogenase deficient, glutamate requiring) background. These mutants, i and en-am, are so-called enhancers of am; they have been redesignated herein as en(am)-1 and en(am)-2, respectively. Although glutamate synthase levels in en(am)-1 were essentially wild type, the en(am)-2 strain was devoid of glutamate synthase activity under all conditions examined, suggesting that en(am)-2 may be the structural locus for glutamate synthase. Regulation of glutamate synthase occurred to some extent, presumably in response to glutamate requirements. Glutamate starvation, as in am mutants, led to enhanced activity. In contrast, glutamine limitation, as in gln-1 mutants, depressed glutamate synthase levels.

scholarly journals The sRNA NsiR4 is involved in nitrogen assimilation control in cyanobacteria by targeting glutamine synthetase inactivating factor IF7

2015 ◽  
Vol 112 (45) ◽  
pp. E6243-E6252 ◽  
Author(s):  
Stephan Klähn ◽  
Christoph Schaal ◽  
Jens Georg ◽  
Desirée Baumgartner ◽  
Gernot Knippen ◽  
...  
Keyword(s):  
Glutamine Synthetase ◽  
Nitrogen Metabolism ◽  
Nitrogen Assimilation ◽  
Nitrogen Sources ◽  
Transcriptome Profiling ◽  
Site Directed Mutagenesis ◽  
Reporter System ◽  
Available Nitrogen ◽  
Mrna Accumulation

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.

scholarly journals Glutamate synthase levels in Neurospora crassa mutants altered with respect to nitrogen metabolism.

1981 ◽  
Vol 1 (2) ◽  
pp. 158-164 ◽  
Author(s):  
N S Dunn-Coleman ◽  
E A Robey ◽  
A B Tomsett ◽  
R H Garrett
Keyword(s):  
Neurospora Crassa ◽  
Glutamate Dehydrogenase ◽  
Nitrogen Metabolism ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Glutamate Synthase ◽  
Wild Type ◽  
Mutant Strains ◽  
Synthase Regulation ◽  
Glutamate Biosynthesis ◽  
Glutamine Limitation

Glutamate synthase catalyzes glutamate formation from 2-oxoglutarate plus glutamine and plays an essential role when glutamate biosynthesis by glutamate dehydrogenase is not possible. Glutamate synthase activity has been determined in a number of Neurospora crassa mutant strains with various defects in nitrogen metabolism. Of particular interest were two mutants phenotypically mute except in an am (biosynthetic nicotinamide adenine dinucleotide phosphate-glutamate dehydrogenase deficient, glutamate requiring) background. These mutants, i and en-am, are so-called enhancers of am; they have been redesignated herein as en(am)-1 and en(am)-2, respectively. Although glutamate synthase levels in en(am)-1 were essentially wild type, the en(am)-2 strain was devoid of glutamate synthase activity under all conditions examined, suggesting that en(am)-2 may be the structural locus for glutamate synthase. Regulation of glutamate synthase occurred to some extent, presumably in response to glutamate requirements. Glutamate starvation, as in am mutants, led to enhanced activity. In contrast, glutamine limitation, as in gln-1 mutants, depressed glutamate synthase levels.

scholarly journals Roles of Glutamate Synthase, gltBD, and gltF in Nitrogen Metabolism of Escherichia coli and Klebsiella aerogenes

2001 ◽  
Vol 183 (22) ◽  
pp. 6607-6619 ◽  
Author(s):  
Thomas J. Goss ◽  
Ana Perez-Matos ◽  
Robert A. Bender
Keyword(s):  
Escherichia Coli ◽  
Nitrogen Metabolism ◽  
Glutamate Synthase ◽  
Nitrogen Sources ◽  
Enteric Bacteria ◽  
The Other ◽  
Klebsiella Aerogenes ◽  
E Coli ◽  
Glutamine Synthesis ◽  
Subsequent Failure

ABSTRACT Mutants of Escherichia coli and Klebsiella aerogenes that are deficient in glutamate synthase (glutamate-oxoglutarate amidotransferase [GOGAT]) activity have difficulty growing with nitrogen sources other than ammonia. Two models have been proposed to account for this inability to grow. One model postulated an imbalance between glutamine synthesis and glutamine degradation that led to a repression of the Ntr system and the subsequent failure to activate transcription of genes required for the use of alternative nitrogen sources. The other model postulated that mutations in gltB or gltD (which encode the subunits of GOGAT) were polar on a downstream gene,gltF, which is necessary for proper activation of gene expression by the Ntr system. The data reported here show that thegltF model is incorrect for three reasons: first, a nonpolar gltB and a polar gltD mutation of K. aerogenes both show the same phenotype; second,K. aerogenes and several other enteric bacteria lack a gene homologous to gltF; and third, mutants of E. coli whose gltF gene has been deleted show no defect in nitrogen metabolism. The argument that accumulated glutamine represses the Ntr system in gltB or gltDmutants is also incorrect, because these mutants can derepress the Ntr system normally so long as sufficient glutamate is supplied. Thus, we conclude that gltB or gltD mutants grow slowly on many poor nitrogen sources because they are starved for glutamate. Much of the glutamate formed by catabolism of alternative nitrogen sources is converted to glutamine, which cannot be efficiently converted to glutamate in the absence of GOGAT activity. Finally, GOGAT-deficient E. coli cells growing with glutamine as the sole nitrogen source increase their synthesis of the other glutamate-forming enzyme, glutamate dehydrogenase, severalfold, but this is still insufficient to allow rapid growth under these conditions.

Control of growth, secondary metabolism and sporulation in Streptomyces venezuelae ISP5230 by jadW 1, a member of the afsA family of γ-butyrolactone regulatory genes

Microbiology ◽  
2003 ◽  
Vol 149 (8) ◽  
pp. 1991-2004 ◽  
Author(s):  
Liru Wang ◽  
Leo C. Vining
Keyword(s):  
Growth Rate ◽  
Secondary Metabolism ◽  
Morphological Differentiation ◽  
Nitrogen Sources ◽  
Repeat Sequence ◽  
Antibiotic Biosynthesis ◽  
Wild Type ◽  
Streptomyces Venezuelae ◽  
New Genes ◽  
Insertional Inactivation

Three new genes (jadW 1, jadW 2 and jadW 3) were isolated from a region of the Streptomyces venezuelae ISP5230 chromosome at the left-hand end of the jad cluster for jadomycin B (JdB) biosynthesis. The deduced amino acid sequence of jadW 1 showed strong similarity to gene products associated in several streptomycetes with γ-butyrolactone autoregulators controlling morphological differentiation and secondary metabolism. Examination of JadW1 for conserved domains detected a repeat sequence characteristic of proteins in the AfsA regulatory family. Insertional inactivation of jadW 1 reduced the growth rate of S. venezuelae cultures in aerated liquid media containing complex nitrogen sources and altered growth morphology in minimal medium. It also affected sporulation on agar media. Cultures of jadW 1-disrupted mutants grown under conditions supporting biosynthesis of JdB or chloramphenicol by the wild-type strain failed to produce either of the antibiotics. Complementing the disrupted strain by transformation with pJV435, containing a cloned copy of the gene, improved sporulation and restored antibiotic biosynthesis in transformants to titres close to those of the wild-type similarly transformed with pJV435 as a control. The results are consistent with a role for jadW 1 in regulating morphological and metabolic differentiation. Further sequence analysis of jadR 2, which functions with jadR 1 in stress-induced activation of JdB biosynthesis, indicated that this gene encodes a γ-butyrolactone receptor homologue. The growth-rate-sensitive phenotype of the jadW 1-disrupted mutant, and the proximity of jadW 1 to jadR 2 indicate that this region of the jad gene cluster contains a regulatory mechanism incorporating γ-butyrolactone signalling and sensitivity to environmental stress.

scholarly journals Post-Silking Shading Stress Affects Leaf Nitrogen Metabolism of Spring Maize in Southern China

Plants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 210 ◽  
Author(s):  
Jue Wang ◽  
Kai Shi ◽  
Weiping Lu ◽  
Dalei Lu
Keyword(s):  
Grain Yield ◽  
Nitrogen Metabolism ◽  
Southern China ◽  
Glutamate Synthase ◽  
Biomass Accumulation ◽  
Leaf Nitrogen ◽  
Dry Matter Accumulation ◽  
Light Deprivation ◽  
Spad Value ◽  
Spring Maize

Lower sunlight caused by overcast skies from June to July in Southern China is one of the main environmental stresses that frequently occur and affect the post-silking growth and grain development of spring maize. In this study, a field trial involving four maize hybrids as materials was conducted to investigate the effects of post-silking shading stress (30% and 50% light deprivation) on leaf nitrogen metabolism and biomass accumulation during maize growing seasons in 2016 and 2017. Results indicated that 30% and 50% shading stress caused the grain yield to decrease by 47.3% and 69.6%, respectively. Plant post-silking biomass accumulation was decreased by shading, whereas the translocation from pre-silking assimilates in the vegetative organs was increased by shading. This change was sharply observed when the plants were deprived of more sunlight intensity. The leaf relative chlorophyll (soil and plant analyzer development (SPAD) value) and soluble protein contents were considerably decreased by shading under 50% light deprivation condition. The activities of nitrate reductase, glutamine synthetase and glutamate synthase that are involved in nitrogen metabolism were downregulated by shading stresses. In conclusion, nitrogen metabolism was disturbed by shading, which induced the decrease in post-silking dry matter accumulation, ultimately resulting in grain yield loss.

scholarly journals Effects of salinity on photosynthetic traits, ion homeostasis and nitrogen metabolism in wild and cultivated soybean

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e8191 ◽  
Author(s):  
Abd Ullah ◽  
Mingxia Li ◽  
Javaria Noor ◽  
Akash Tariq ◽  
Yuan Liu ◽  
...  
Keyword(s):  
Glutamate Dehydrogenase ◽  
Nitrogen Metabolism ◽  
Nitrogen Assimilation ◽  
Wild Soybean ◽  
Nitrate Assimilation ◽  
Saline Stress ◽  
Alkaline Stress ◽  
Ion Regulation ◽  
Cultivated Soybean ◽  
The Impact

Background Carbon and nitrogen metabolism need to be highly regulated to achieve cell acclimation to changing environmental conditions. The understanding of physio-biochemical responses of crops to salinity stress could help to stabilize their performance and yield. In this study we have analyzed the roles of photosynthesis, ion physiology and nitrate assimilation toward saline/alkaline stress acclimation in wild and cultivated soybean seedlings. Methods Growth and photosynthetic parameters, ion concentrations and the activity of enzymes involved in nitrogen assimilation were determined in seedlings of one wild and one cultivated soybean accession subjected to saline or alkaline stresses. Results Both saline and alkaline stresses had a negative impact on the growth and metabolism of both wild and cultivated soybean.The growth, photosynthesis, and gas exchange parameters showed a significant decrease in response to increasing salt concentration. Additionally, a significant increase in root Na+ and Cl– concentration was observed. However, photosynthetic performance and ion regulation were higher in wild than in cultivated soybean under saline and alkaline stresses. Nitrate reductase (NR) and the glutamine synthetase/glutamate synthase (GS/GOGAT) cycle showed a significant decrease in leaves of both genotypes. The reduction in the GS/GOGAT cycle was accompanied by high aminating glutamate dehydrogenase (NADH-glutamate dehydrogenase) activity, indicating the assimilation of high levels of NH4+. A significant increase in the activities of aminating and deaminating enzymes, including glutamate dehydrogenase (GDH), alanine aminotransferase (AlaAT) and aspartate aminotransferase (AspAT), was observed, probably due to the high glutamate demand and maintenance of the Krebs cycle to correct the C: N status. Conclusions Cultivated soybean was much more stress sensitive than was the wild soybean. The decrease in growth, photosynthesis, ion regulation and nitrogen assimilation enzymes was greater in cultivated soybean than in wild soybean. The impact of alkaline stress was more pronounced than that of saline stress. Wild soybean regulated the physiological mechanisms of photosynthesis and nitrate assimilation more effectively than did cultivated soybean. The present findings provide a theoretical basis with which to screen and utilize wild and cultivated soybean germplasm for breeding new stress-tolerant soybean.

scholarly journals The roles of glutamate dehydrogenase, glutamine synthetase and three forms of glutamate synthase on nitrogen assimilation in various organs of Pisum arvense L.

2014 ◽  
Vol 60 (3-4) ◽  
pp. 295-302
Author(s):  
Genowefa Kubik-Dobosz
Keyword(s):  
Glutamine Synthetase ◽  
Glutamate Dehydrogenase ◽  
Nitrogen Assimilation ◽  
Decisive Role ◽  
Glutamate Synthase ◽  
Pisum Arvense ◽  
Age Dependent ◽  
The Individual

The activities of GDH, GS and three forms of GOGAT (NADH, NADPH or ferredoxin-dependent) were studied in the leaves, stems and roots of the <i>Pisum arvense</i>. GS and the individual forms of GOGAT dominated in the leaves of 7 day-old plants which were taking up NO<sub>3</sub><sup>-</sup> or NH<sub>4</sub><sup>+</sup> ions, while NADH-GDH dominated in the roots of these plants. In comparison with HNO<sub>3</sub><sup>-</sup> , NH<sub>4</sub><sup>+</sup> ions stimulated the activity of most of the enzymes of the GS/GOGAT and GDH pathways in stems and roots, while in and leaves this effect was age-dependent. The Fd-GOGAT located in leaves and stems was not regulated by NH<sub>4</sub><sup>+</sup> , which indicates that this enzyme is not likely to be directly involved in the assimilation of NH<sub>4</sub><sup>+</sup> ions that have been taken up. The obtained data indicate that at lower tissue NH<sub>4</sub><sup>+</sup> concentration a decisive role in nitrogen assimilation in leaves and stems is played by the GS/GOGAT pathway, while in the roots-by GDH and in less degree by GS, GOGAT. High amounts of accumulated NH<sub>4</sub><sup>+</sup> ions set off a detoxication mechanism which includes NADH-GDH, common to all tissues. Only in 7 day-old leaves did the detoxication of NH<sub>4</sub><sup>+</sup> take place with the involvement of NADH-GOGAT and NADPH-GOGAT.

Effect of ammonium on chloramphenicol production by Streptomyces venezuelae in batch and continuous cultures

1985 ◽  
Vol 31 (2) ◽  
pp. 119-123 ◽  
Author(s):  
S. Shapiro ◽  
L. C. Vining
Keyword(s):  
Nitrogen Assimilation ◽  
Specific Growth ◽  
Antibiotic Production ◽  
Nitrogen Sources ◽  
Streptomyces Venezuelae ◽  
Continuous Cultures ◽  
Limiting Nutrient ◽  
Nitrogen Repression ◽  
Specific Growth Rates ◽  
Temporary Inhibition

Cultures of Streptomyces venezuelae presented with a mixture of ammonium and an amino acid as nitrogen sources used both compounds together. Absence of ammonium repression of alternative nitrogen assimilation pathways was also observed when ammonium was added to cultures already growing on proline. The presence of ammonium in the medium ab initio depressed the yield of chloramphenicol. However, its addition to a culture growing on proline caused only a temporary inhibition of antibiotic synthesis, even when sufficient ammonium was added to create an excess. Continuous cultures supplied with ammonium as the growth-limiting nutrient showed no significant change in specific antibiotic production at different specific growth rates. The overall results indicate that in S. venezuelae neither nitrogen utilization pathways nor chloramphenicol biosynthesis is controlled by nitrogen repression.

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