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ATP + [L-Asp(4-L-Arg)]n + alpha-L-Asp-L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n + beta-L-Asp-L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
[L-Asp(4-L-Arg)]n+1 + ADP + phosphate
L-Lys cannot replace L-Arg in reaction with CphANE1 or CphANE1del96
-
-
?
L-aspartic acid + ATP
poly-L-aspartic acid + ADP + phosphate
additional information
?
-
ATP + [L-Asp(4-L-Arg)]n + beta-L-Asp-L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n + beta-L-Asp-L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
study of the effect of different light and nutrition conditions on cyanophycin granule formation
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
role of the C-terminal region of CphANE1, Glu856 is critical for CphANE1 catalytic activity, overview
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
role of the C-terminal region of CphANE1, Glu856 is critical for CphANE1 catalytic activity, overview
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
study of expression profiles of the genes cphA1 and cphA2 and their dependence on the type of nitrogen supply in the medium
[L-Asp(4-L-Arg)]n-Asp is the substrate for the second reaction catalysed by cyanophycin synthase
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
a small amount of cyanophycin is required as a primer
no formation of AMP, [L-Asp(3-L-Arg)]n-Asp is the substrate for the second reaction catalysed by cyanophycin synthase
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
normal assay conditions
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
Thermosynechococcus vestitus
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
Thermosynechococcus vestitus
-
primer and product analysis using recombinantly expressed Tlr2170 protein, overview
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
a small amount of cyanophycin is required as a primer
no formation of AMP, [L-Asp(4-L-Arg)]n-Asp is the substrate for the second reaction catalysed by cyanophycin synthase
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
a small amount of cyanophycin is required as a primer
no formation of AMP, [L-Asp(4-L-Arg)]n-Asp is the substrate for the second reaction catalysed by cyanophycin synthase, EC 6.3.2.30
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
a short peptide is required as a primer, the primer peptide is elongated at its C-terminus
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
L-aspartic acid + ATP
poly-L-aspartic acid + ADP + phosphate
-
-
-
-
?
L-aspartic acid + ATP
poly-L-aspartic acid + ADP + phosphate
-
-
-
?
L-aspartic acid + ATP
poly-L-aspartic acid + ADP + phosphate
-
-
-
-
?
additional information
?
-
-
without L-aspartic acid 9.2% activity compared to the reaction mixture containing both substrates, no activity using L-glutamic acid instead of L-aspartic acid, no activity without addition of small amounts of cyanophycin as a primer for synthesis
-
-
?
additional information
?
-
cyanophycin accumulation is studied, comparison of nitrogen-fixing and non-nitrogen-fixing cyanobacteria
-
-
?
additional information
?
-
-
the enzyme catalyzes the two-step reaction performing reactions of EC 6.3.2.30 and EC 6.3.2.29
-
-
?
additional information
?
-
-
the enzyme catalyzes the two-step reaction performing reactions of EC 6.3.2.30 and EC 6.3.2.29
-
-
?
additional information
?
-
negligible activities when L-aspartic acid is omitted
-
-
?
additional information
?
-
no activity without L-aspartic acid, L-glutamic acid cannot substitute for L-aspartic acid
-
-
?
additional information
?
-
Thermosynechococcus vestitus
-
CphA requires L-Asp, L-Arg, ATP, Mg2+, and low-molecular mass cyanophycin as a primer for cyanophycin synthesis and catalyzes the elongation of a low-molecular mass cyanophycin, overview
-
-
?
additional information
?
-
Thermosynechococcus vestitus
-
the enzyme catalyzes the two-step reaction performing reactions of EC 6.3.2.30 and EC 6.3.2.29. Recombinant Tlr2170 protein catalyzes in vitro cyanophycin synthesis in the absence of a primer. The Tlr2170 protein shows strict substrate specificity toward L-Asp and L-Arg
-
-
?
additional information
?
-
-
no product is formed if the peptide primer is blocked at the C-terminus or if L-arginine is solely supplied as substrate
-
-
?
additional information
?
-
enzyme is specific for the dipeptide beta-aspartyl-arginine and ATP as substrates. To incorporate 1 mol beta-aspartyl-arginine into cyanophycin, the hydrolysis of about 1 mol ATP is required. No substrates: beta-aspartyl-lysine, alpha-aspartyl-glycine, beta-aspartyl-glycine, alpha-aspartyl-leucine, beta-aspartyl-leucine, beta-aspartyl-alanine, beta-aspartyl-phenylalanine, alpha-glutamyl-leucine, alpha-alanyl-glycine, L-aspartate, L-arginine
-
-
?
additional information
?
-
cyanophycin synthetase catalyzes the synthesis of cyanophycin by ATP-dependent polymerization of L-Asp and L-Arg. In vitro, the activity of CphA generally depends on the presence of L-Asp, L-Arg, ATP, Mg2+, K+, sulfhydryl compound, and cyanophycin as primers
-
-
?
additional information
?
-
the recombinant CphA49 exhibits strict primer dependency and broad substrate specificities. L-Lys and L-Glu can substitute for L-Arg, but with very low catalytic activity. No activity with L-Arg and L-citrulline, L-Arg and L-ornithine, L-Asp and L-citrulline, and L-Asp and L-ornithine
-
-
?
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ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
additional information
?
-
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
study of expression profiles of the genes cphA1 and cphA2 and their dependence on the type of nitrogen supply in the medium
[L-Asp(4-L-Arg)]n-Asp is the substrate for the second reaction catalysed by cyanophycin synthase
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
Thermosynechococcus vestitus
-
-
-
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
a small amount of cyanophycin is required as a primer
no formation of AMP, [L-Asp(4-L-Arg)]n-Asp is the substrate for the second reaction catalysed by cyanophycin synthase
-
?
ATP + [L-Asp(4-L-Arg)]n-L-Asp + L-Arg
ADP + phosphate + [L-Asp(4-L-Arg)]n+1
-
-
-
?
additional information
?
-
-
without L-aspartic acid 9.2% activity compared to the reaction mixture containing both substrates, no activity using L-glutamic acid instead of L-aspartic acid, no activity without addition of small amounts of cyanophycin as a primer for synthesis
-
-
?
additional information
?
-
Thermosynechococcus vestitus
-
CphA requires L-Asp, L-Arg, ATP, Mg2+, and low-molecular mass cyanophycin as a primer for cyanophycin synthesis and catalyzes the elongation of a low-molecular mass cyanophycin, overview
-
-
?
additional information
?
-
cyanophycin synthetase catalyzes the synthesis of cyanophycin by ATP-dependent polymerization of L-Asp and L-Arg. In vitro, the activity of CphA generally depends on the presence of L-Asp, L-Arg, ATP, Mg2+, K+, sulfhydryl compound, and cyanophycin as primers
-
-
?
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E856A
-
site-directed mutagenesis, inactive mutant
E856V
-
site-directed mutagenesis, inactive mutant
E856A
-
site-directed mutagenesis, inactive mutant
-
E856V
-
site-directed mutagenesis, inactive mutant
-
C595S
enhanced catalytic activity
C595S
-
site-directed mutagenesis, the mutant shows a 2.5fold increased catalytic activity compared to the wild-type enzyme
C595S
-
enhanced catalytic activity
-
additional information
two truncated CphAs, lacking 31 (CphANE1del96) or 59 (CphANE1del180) amino acids of the C-terminal region, are derived from cphANE1 by deleting 96 or 180 bp from its 3' region through the introduction of stop codons. In comparison to the wild-type gene, cphANE1del96 conferrs about 2.1-2.2fold higher enzyme activity on Escherichia coli. These cells accumulate about twofold more cyanophycin than cells expressing the wild-type gene
additional information
-
two truncated CphAs, lacking 31 (CphANE1del96) or 59 (CphANE1del180) amino acids of the C-terminal region, are derived from cphANE1 by deleting 96 or 180 bp from its 3' region through the introduction of stop codons. In comparison to the wild-type gene, cphANE1del96 conferrs about 2.1-2.2fold higher enzyme activity on Escherichia coli. These cells accumulate about twofold more cyanophycin than cells expressing the wild-type gene
additional information
-
construction of truncation mutant CphANE1-C45, which lacks up to 45 amino acids at its C-terminus, the mutant retains full enzymatic activity and produced polymers, however, the removal of one additional amino acid, Glu856, results in complete inactivation of CphANE1-C46, phenotype, overview. Removal of the sequence Leu867-Leu870 results in dramatically decreased thermostability
additional information
-
construction of truncation mutant CphANE1-C45, which lacks up to 45 amino acids at its C-terminus, the mutant retains full enzymatic activity and produced polymers, however, the removal of one additional amino acid, Glu856, results in complete inactivation of CphANE1-C46, phenotype, overview. Removal of the sequence Leu867-Leu870 results in dramatically decreased thermostability
-
additional information
-
construction of deletion mutants CphA6308DELTA1, CphA6308DELTA2, and CphA6308DELTA3, which are truncated by one, two, or three amino acid(s), respectively, at the C-terminus, and a combination of the deletion mutation with point mutation C595S, mutants CphA6308DELTA1 and CphA6308DELTA1/C595S show increased catalytic activity compared to the wild-type enzyme, while mutants CphA6308DELTA2 and CphA6308DELTA3 show similar or reduced CphA enzyme activity in comparison to wild-type CphA6308
additional information
Thermosynechococcus vestitus
-
generation of transgenic Solanum tuberosum plants that recombinantly produce the bacterial enzyme by tuber-specific expression. The mutant plants show increased cyanophycin contents, and the tuber-specific expression minimizes the phenotypic abnormalities of the sprout caused by cyanophycin accumulation, phenotypes,overview
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engineered cyanophycin synthetase (CphA) from Nostoc ellipsosporum confers enhanced CphA activity and cyanophycin accumulation to Escherichia coli
expression in Escherichai coli
expression in Escherichia coli
expression in Escherichia coli BL21(DE3)
-
expression in Escherichia coli DH1 cells
-
expression in Escherichia coli DH5alpha
expression in Escherichia coli TOP10 cells
expression in Pseudomonas putida ATCC 4359
expression in the wild-type Sinorhizobium meliloti 1021 and in a phbC-negative mutant
-
functional expression in Nicotiana tabacum var. Petit Havana SRI targeted to the chloroplasts using the CaMV 35S promoter and a translocation pathway signal sequence, the phenotypic abnormalities are reduced by this way, cyanophycin accumulation in chloroplasts, overview
Thermosynechococcus vestitus
-
gene cphA, DNA and amino acid sequence determination and analysis, phylogenetic analysis, functional overexpression of His-tagged Tlr2170 in Escherichia coli BL21(DE3)
Thermosynechococcus vestitus
-
gene cphA, expression in Solanum tuberosum tubers under control of the tuber-specific class 1 promoter B33 directed to tuber cytosol or to tuber plastids leading to cyanophycin accumulation in the tubers
Thermosynechococcus vestitus
-
gene cphA, subcloning and expression of His-tagged wild-type and mutant enzymes in Escherichia coli strains DH5alpha and BL21(DE3)
-
gene cphA49 cloned from deep-sea sediment metagenomic library with degenerated primers, amplified from the fss49 fosmid by PCR, DNA and amino acid sequence determination and analysis, functional expression of His-tagged cphA49 in Escherichia coli strain BL21(DE3), phylogenetic tree, subcloning in Escherichia coli strain DH5alpha
gene cphA6308, expression in Pichia pastoris and Escherichia coli from expression vectors pPICHOLI-C with the copper-inducible CUP1 promoter and pPICHOLI-3 with the methanol-inducible AOX1 promoter. Stabilization of the expression plasmid, the his4 gene from Saccharomyces cerevisiae is introduced and the expression is made in the His auxotrophic Pichia pastoris strain GS115. Recombinant polymer production by wild-type and mutant enzymes and product compositions, overview
-
gene cphA6308, functional expression in Saccharomyces cerevisiae strains G175 and BY4741, which is much more efficient with the copper ion-inducible CUP1 promoter instead of the GAL1 promoter, the yeast strains produce water-soluble and water-insoluble cyanophycin polymer. Growth of transgenic yeasts in the presence of 15 mM lysine results in an incorporation of up to 10 mol% of lysine into cyanophycin, overview. Subcloning in Escherichia coli strain XL1-Blue
-
gene cphA6803, enzyme expression in auxotrophic mutant Rhizopus oryzae strain M16 under control of the pyruvate decarboxylase promoter and terminator elements of Rhizopus oryzae by biolistic transformation method
gene cphA7120, enzyme expression in auxotrophic mutant Rhizopus oryzae strain M16 under control of the pyruvate decarboxylase promoter and terminator elements of Rhizopus oryzae by biolistic transformation method
expression in Escherichia coli
-
expression in Escherichia coli
-
expression in Escherichia coli TOP10 cells
expression in Escherichia coli TOP10 cells
-
expression in Escherichia coli TOP10 cells
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synthesis
-
establishment of cyanophycin, i.e. multi-L-arginylpoly-L-aspartic acid or CGP, synthesis and applicability of this industrially widely used microorganism for the production of this polyamide, overview
synthesis
Thermosynechococcus vestitus
-
production of biodegradable polymers that can be used to substitute petrochemical compounds in commercial products by the recombinant enzyme in transgenic plants
synthesis
-
expression of cyanophycin synthase in wild-type Sinorhizobium meliloti 1021 and in a phbC-negative mutant. Yeast mannitol broth yields the highest cyanophycin contents in both Sinorhizobium meliloti 1021 strains. Supplying the medium with isopropyl-beta-D-thiogalactopyranoside, aspartic acid, and arginine enhances cyanophycin contents about 2.5- and 2.8fold. Varying the nitrogen-to-carbon ratio in the medium enhanced the cyanophycin content further to 43.8% w/w of cell dry weight. Cyanophycin from the Sinorhizobium meliloti strains consists of equimolar amounts of aspartic acid and arginine and contains no other amino acids even if the medium is supplemented with glutamic acid, citrulline, ornithine, or lysine. Cyanophycin isolated from Sinorhizobium meliloti exhibits average molecular weights between 20 and 25 kDa. Cyanophycin isolated after expression in Escherichia coli S17-1 exhibits average molecular weight between 22 and 30 kDa. Co-expression of cyanophycinase from Anabaena sp. PCC7120 encoded by cphB17120 in cphA17120-positive Escherichia coli S17-1, Sinorhizobium meliloti 1021, and its phbC-negative mutant gives cyanophycinase activities in crude extracts, and no CGP granules occur
synthesis
expression of the cyanophycin synthetase of Synechocystis sp. PCC 6308 in Pseudomonas putida ATCC 4359 using an optimised medium for cultivation, results in synthesis of insoluble cyanophycin up to 14.7 w/w and soluble cyanophycin amounting up to 28.7 w/w of the cell dry matter. The soluble CGP is composed of 50.4 mol% aspartic acid, 32.7 mol% arginine, 8.7 mol% citrulline and 8.3 mol% lysine. The insoluble cyanophycin contains less than 1 mol% of citrulline. Using a mineral salt medium with 1.25 or 2% w/v sodium succinate, respectively, plus 23.7 mM L-arginine, the cells synthesise insoluble cyanophycin amounting up to 25% to 29% of the CDM with only a very low citrulline content
synthesis
Thermosynechococcus vestitus
-
restriction of cyanophycin accumulation to the potato tubers by using the cyanophycin synthetase gene from Thermosynechococcus elongatus BP-1, under the control of the tuber-specific class 1 promoter. Tuber-specific cytosolic expression by pB33-cphATe as well as tuber-specific plastidic expression by pB33-PsbYcphATe results in significant polymer accumulation solely in the tubers. In plants transformed with pB33-cphATe, both cyanophycin synthetase and cyanophycin are detected in the cytoplasm leading to an increase up to 2.3% cyanophycin of dry weight and resulting in small and deformed tubers. In B33-PsbY-cphATe tubers, cyanophycin synthetase and cyanophycin are exclusively found in amyloplasts leading to a cyanophycin accumulation up to 7.5% of dry weight. These tubers are normal in size, some clones show reduced tuber yield and sometimes exhibit brown sunken staining starting at tubers navel. During a storage period over of 32 weeks of one selected clone, the cyanophycin content was stable in B33-PsbYcphATe tubers but the stress symptoms increased. Nitrogen fertilization in the greenhouse does not lead not to an increased cyanophycin yield, slightly reduced protein content, decreased starch content, and changes in the amounts of bound and free arginine and aspartate
synthesis
synthesis of cyanophycin using an anabolism-based media-dependent plasmid addiction system to enhance plasmid stability, and a process based on a modified mineral salts medium yielding a cyanophycin content of 42% w/w at the maximum without the addition of amino acids to the medium. This plasmid addiction system is based on different lysine biosynthesis pathways and consists of a knockout of the chromosomal dapE that disrupts the native succinylase pathway in Escherichia coli and the complementation by the plasmid-encoded artificial aminotransferase pathway mediated by the dapL gene from Synechocystis sp. PCC 6308, which allows the synthesis of the essential lysine precursor L,L-2,6-diaminopimelate. This plasmid also harbors an engineered cyanophycin synthetase gene responsible for cyanophycin production.Cultivation experiments reveal an at least 4.5fold enhanced production of cyanophycin in comparison to control cultivations
synthesis
-
cyanophycin produced in Escherichia coli is composed of 50% of aspartic acid, 45% of arginine, and 3.5% of lysine, and exhibits a homogenous molecular mass of 35 kDa. Cultivation in presence of arginine, aspartic acid, lysine and glucose with the minimal resource leads to 1.72 g/l soluble cyanophycin
synthesis
-
expression of cyanophycin synthase in wild-type Sinorhizobium meliloti 1021 and in a phbC-negative mutant. Yeast mannitol broth yields the highest cyanophycin contents in both Sinorhizobium meliloti 1021 strains. Supplying the medium with isopropyl-beta-D-thiogalactopyranoside, aspartic acid, and arginine enhances cyanophycin contents about 2.5- and 2.8fold. Varying the nitrogen-to-carbon ratio in the medium enhanced the cyanophycin content further to 43.8% w/w of cell dry weight. Cyanophycin from the Sinorhizobium meliloti strains consists of equimolar amounts of aspartic acid and arginine and contains no other amino acids even if the medium is supplemented with glutamic acid, citrulline, ornithine, or lysine. Cyanophycin isolated from Sinorhizobium meliloti exhibits average molecular weights between 20 and 25 kDa. Cyanophycin isolated after expression in Escherichia coli S17-1 exhibits average molecular weight between 22 and 30 kDa. Co-expression of cyanophycinase from Anabaena sp. PCC7120 encoded by cphB17120 in cphA17120-positive Escherichia coli S17-1, Sinorhizobium meliloti 1021, and its phbC-negative mutant gives cyanophycinase activities in crude extracts, and no CGP granules occur
-
synthesis
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synthesis of cyanophycin using an anabolism-based media-dependent plasmid addiction system to enhance plasmid stability, and a process based on a modified mineral salts medium yielding a cyanophycin content of 42% w/w at the maximum without the addition of amino acids to the medium. This plasmid addiction system is based on different lysine biosynthesis pathways and consists of a knockout of the chromosomal dapE that disrupts the native succinylase pathway in Escherichia coli and the complementation by the plasmid-encoded artificial aminotransferase pathway mediated by the dapL gene from Synechocystis sp. PCC 6308, which allows the synthesis of the essential lysine precursor L,L-2,6-diaminopimelate. This plasmid also harbors an engineered cyanophycin synthetase gene responsible for cyanophycin production.Cultivation experiments reveal an at least 4.5fold enhanced production of cyanophycin in comparison to control cultivations
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synthesis
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expression of the cyanophycin synthetase of Synechocystis sp. PCC 6308 in Pseudomonas putida ATCC 4359 using an optimised medium for cultivation, results in synthesis of insoluble cyanophycin up to 14.7 w/w and soluble cyanophycin amounting up to 28.7 w/w of the cell dry matter. The soluble CGP is composed of 50.4 mol% aspartic acid, 32.7 mol% arginine, 8.7 mol% citrulline and 8.3 mol% lysine. The insoluble cyanophycin contains less than 1 mol% of citrulline. Using a mineral salt medium with 1.25 or 2% w/v sodium succinate, respectively, plus 23.7 mM L-arginine, the cells synthesise insoluble cyanophycin amounting up to 25% to 29% of the CDM with only a very low citrulline content
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uncultured bacterium (M9UYB0)
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Clostridium perfringens (Q0SQX0), Clostridium perfringens, Clostridium perfringens SM101 (Q0SQX0)
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unidentified microorganism
brenda
Klemke, F.; Nuernberg, D.J.; Ziegler, K.; Beyer, G.; Kahmann, U.; Lockau, W.; Volkmer, T.
CphA2 is a novel type of cyanophycin synthetase in N2-fixing cyanobacteria
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Cyanothece sp. PCC 7425, Trichormus variabilis (Q3MGC5)
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Thermosynechococcus vestitus
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