Engineered Cyanophycin Synthetase (CphA) from
            <i>Nostoc ellipsosporum</i>
            Confers Enhanced CphA Activity and Cyanophycin Accumulation to
            <i>Escherichia coli</i>

Hải, Trần Ngọc; Frey, Kay; Steinbüchel, Alexander

doi:10.1128/aem.01132-06

Cited by 24 publications

(18 citation statements)

References 46 publications

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“…Reduced ppGpp levels would result in lower levels of expression of FevRdependent genes. The cphA gene is implicated in creating amino acid storage molecules (equal parts aspartic acid and arginine) in cyanobacteria (26). While there is no evidence that cyanophycin is produced by Francisella strains, one hypothesis for the explanation of the phenotypes of this F. tularensis mutant, without more specific information, is that deletion of cphA increases the availability of amino acids for growth, reducing the production of ppGpp and genes activated by high levels of the alarmone.…”

Section: Discussionmentioning

confidence: 99%

“…The F. tularensis CphA protein is ϳ50% similar to the CphA protein of cyanobacteria, which is involved in making the nitrogen reserve molecule cyanophycin (i.e., granules of arginine, lysine, or asparagine) (26). It is unknown if Francisella produces cyanophycin; however, the genome of Francisella does not appear to contain cphB, which is a cyanophycinase (27).…”

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confidence: 99%

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The Francisella tularensis migR, trmE , and cphA Genes Contribute to F. tularensis Pathogenicity Island Gene Regulation and Intracellular Growth by Modulation of the Stress Alarmone ppGpp

et al. 2013

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fThe Francisella tularensis pathogenicity island (FPI) encodes many proteins that are required for virulence. Expression of these genes depends upon the FevR (PigR) regulator and its interactions with the MglA/SspA and RNA polymerase transcriptional complex. Experiments to identify how transcription of the FPI genes is activated have led to identification of mutations within the migR, trmE, and cphA genes that decrease FPI expression. Recent data demonstrated that the small alarmone ppGpp, produced by RelA and SpoT, is important for stabilizing MglA/SspA and FevR (PigR) interactions in Francisella. Production of ppGpp is commonly known to be activated by cellular and nutritional stress in bacteria, which indicates that cellular and nutritional stresses act as important signals for FPI activation. In this work, we demonstrate that mutations in migR, trmE, or cphA significantly reduce ppGpp accumulation. The reduction in ppGpp levels was similar for each of the mutants and correlated with a corresponding reduction in iglA reporter expression. In addition, we observed that there were differences in the ability of each of these mutants to replicate within various mammalian cells, indicating that the migR, trmE, and cphA genes are likely parts of different cellular stress response pathways in Francisella. These results also indicate that different nutritional and cellular stresses exist in different mammalian cells. This work provides new information to help understand how Francisella regulates its virulence genes in response to host cell environments, and it contributes to our growing knowledge of this highly successful bacterial pathogen.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

The Francisella tularensis migR, trmE , and cphA Genes Contribute to F. tularensis Pathogenicity Island Gene Regulation and Intracellular Growth by Modulation of the Stress Alarmone ppGpp

et al. 2013

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“…Of particular interest for quantitative proteomics are SI-labeled amino acids, especially SI-arginine, which are used for labeling of proteins in cultured cells, i.e., for the SILAC (stableisotope labeling with amino acids in cell culture) method (30,31). The beauty of this method is that cells growing on 13 C/ 15 N-labeled amino acids incorporate the label into their proteins without any perturbing effects on their metabolism. The production of 13 Clabeled substances using algae and other autotrophic organisms that utilize 13 CO 2 as the sole carbon source is far more cost-effective than using heterotrophic organisms.…”

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confidence: 99%

“…The beauty of this method is that cells growing on 13 C/ 15 N-labeled amino acids incorporate the label into their proteins without any perturbing effects on their metabolism. The production of 13 Clabeled substances using algae and other autotrophic organisms that utilize 13 CO 2 as the sole carbon source is far more cost-effective than using heterotrophic organisms. Not only is 13 CO 2 per se cheaper than 13 C-labeled sugars, but heterotrophs incorporate only a fraction of the label, the rest being lost due to respiration.…”

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Autotrophic Production of Stable-Isotope-Labeled Arginine in Ralstonia eutropha Strain H16

Lütte

Pohlmann

Zaychikov

et al. 2012

Appl Environ Microbiol

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cWith the aim of improving industrial-scale production of stable-isotope (SI)-labeled arginine, we have developed a system for the heterologous production of the arginine-containing polymer cyanophycin in recombinant strains of Ralstonia eutropha under lithoautotrophic growth conditions. We constructed an expression plasmid based on the cyanophycin synthetase gene (cphA) of Synechocystis sp. strain PCC6308 under the control of the strong P cbbL promoter of the R. eutropha H16 cbb c operon (coding for autotrophic CO 2 fixation). In batch cultures growing on H 2 and CO 2 as sole sources of energy and carbon, respectively, the cyanophycin content of cells reached 5.5% of cell dry weight (CDW). However, in the absence of selection (i.e., in antibiotic-free medium), plasmid loss led to a substantial reduction in yield. We therefore designed a novel addiction system suitable for use under lithoautotrophic conditions. Based on the hydrogenase transcription factor HoxA, this system mediated stabilized expression of cphA during lithoautotrophic cultivation without the need for antibiotics. The maximum yield of cyanophycin was 7.1% of CDW. To test the labeling efficiency of our expression system under actual production conditions, cells were grown in 10-liter-scale fermentations fed with 13

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Ubiquitous Acinetobacter species as beneficial commensals but gradually being emboldened with antibiotic resistance genes

Adegoke

Tom

Okoh

2012

J. Basic Microbiol.

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Acinetobacter spp. are ubiquitous obligate aerobic bacteria which occur mostly as commensals on the skin, and in soil, water and plants' rhizosphere. Though the species in this genus have been implicated as aetiologies in some nosocomial infections, their versatility covers biodegradation or dissolution leading to bioremediation; catalysis leading to synthesis of high molecular weight, life sustaining polymers; and as growth enhancers in agriculture. The challenge of antibiotic resistance and their mediatory genes is a cause for concern but should not deter the beneficial application of the bacteria especially in the synthesis of novel compounds that would be of relevance in overcoming some global ecological challenges. This review addresses important beneficial attributes of Acinetobacter species as well as gives some insight into emerging trends in their resistance to antibiotics.

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Engineered Cyanophycin Synthetase (CphA) from Nostoc ellipsosporum Confers Enhanced CphA Activity and Cyanophycin Accumulation to Escherichia coli

Cited by 24 publications

References 46 publications

The Francisella tularensis migR, trmE , and cphA Genes Contribute to F. tularensis Pathogenicity Island Gene Regulation and Intracellular Growth by Modulation of the Stress Alarmone ppGpp

The Francisella tularensis migR, trmE , and cphA Genes Contribute to F. tularensis Pathogenicity Island Gene Regulation and Intracellular Growth by Modulation of the Stress Alarmone ppGpp

Autotrophic Production of Stable-Isotope-Labeled Arginine in Ralstonia eutropha Strain H16

Ubiquitous Acinetobacter species as beneficial commensals but gradually being emboldened with antibiotic resistance genes

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