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show all sequences of 2.1.1.156

Isolation and functional characterization of N-methyltransferases that catalyze betaine synthesis from glycine in a halotolerant photosynthetic organism Aphanothece halophytica

Waditee, R.; Tanaka, Y.; Aoki, K.; Hibino, T.; Jikuya, H.; Takano, J.; Takabe, T.; J. Biol. Chem. 278, 4932-4942 (2003)

Data extracted from this reference:

Activating Compound
Activating Compound
Commentary
Organism
Structure
additional information
abiotic stess-inducible enzyme, e.g. high salt content
Aphanothece halophytica
Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli BL21(DE3); orf1, DNA and amino acid sequence determination and analysis, phylogenetic analysis, overexpression of wild-type and mutant enzymes in Escherichia coli, functional coexpression of the soluble enzyme with the sarcosine dimethylglycine methyltransferase, EC 2.1.1.157, increases the intracellular betaine level and the growth rate of Escherichia coli strain BL21(DE3)
Aphanothece halophytica
Engineering
Amino acid exchange
Commentary
Organism
R169E
inactive; site-directed mutagenesis, completely inactive mutant
Aphanothece halophytica
R169K
drastic increase of Km of the methyl-acceptor substrates, 2-fold increase in Km for S-adenosyl-L-methionine; site-directed mutagenesis, reduced activity and highly increased Km for ATP with both glycine and sarcosine compared to the wild-type enzyme
Aphanothece halophytica
Inhibitors
Inhibitors
Commentary
Organism
Structure
acetate
; 80% inhibition at 250 mM, with glycine
Aphanothece halophytica
dimethylglycine
versus glycine, 65% inhibition at 250 mM
Aphanothece halophytica
ethanolamine
; 45% inhibition at 250 mM, with glycine
Aphanothece halophytica
KCl
-
Aphanothece halophytica
monomethyl ethanolamine
; 35% inhibition at 250 mM, with glycine
Aphanothece halophytica
additional information
inhibition rates with sarcosine, overview, no feedback inhibition by betaine up to 2 M, no inhibition with glycine by serine, methionine, asparagine, glutamate, cysteine, and alanine; no inhibition with betaine
Aphanothece halophytica
N,N-dimethylglycine
-
Aphanothece halophytica
NaCl
-
Aphanothece halophytica
phenylalanine
10% inhibition at 250 mM, with glycine
Aphanothece halophytica
proline
; 30% inhibition at 250 mM, with glycine
Aphanothece halophytica
S-adenosyl-L-homocysteine
; product inhibition
Aphanothece halophytica
sarcosine
competitive inhibition, versus glycine, 70% inhibition at 250 mM
Aphanothece halophytica
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.6
-
S-adenosyl-L-methionine
pH 8.8, 37°C, recombinant wild-type enzyme, with sarcosine; with N-methylglycine as methyl group acceptor, pH 8.8, 37°C
Aphanothece halophytica
0.7
-
S-adenosyl-L-methionine
pH 8.8, 37°C, recombinant wild-type enzyme, with glycine; with glycine as methyl group acceptor, pH 8.8, 37°C
Aphanothece halophytica
0.8
-
N-methylglycine
pH 8.8, 37°C
Aphanothece halophytica
0.8
-
sarcosine
pH 8.8, 37°C, recombinant wild-type enzyme
Aphanothece halophytica
1
-
glycine
pH 8.8, 37°C, recombinant mutant R169K
Aphanothece halophytica
1.1
-
sarcosine
pH 8.8, 37°C, recombinant mutant R169K
Aphanothece halophytica
1.5
-
glycine
pH 8.8, 37°C; pH 8.8, 37°C, recombinant wild-type enzyme
Aphanothece halophytica
50
-
S-adenosyl-L-methionine
pH 8.8, 37°C, recombinant mutant R169K, with sarcosine
Aphanothece halophytica
75
-
S-adenosyl-L-methionine
pH 8.8, 37°C, recombinant mutant R169K, with glycine
Aphanothece halophytica
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
31210
-
calculated from the deduced amino acid sequence
Aphanothece halophytica
32000
-
gel filtration; recombinant enzyme, gel filtration
Aphanothece halophytica
33000
-
1 * 33000, recombinant enzyme, SDS-PAGE; 1 * 33000, SDS-PAGE, 1 * 32000, gel filtration
Aphanothece halophytica
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
additional information
Aphanothece halophytica
the enzyme is involved in the alternate biosynthesis of betaine, which is an important osmoprotectant and is synthesized in response to abiotic stress
?
-
-
-
S-adenosyl-L-methionine + glycine
Aphanothece halophytica
-
S-adenosyl-L-homocysteine + sarcosine
-
-
?
S-adenosyl-L-methionine + glycine
Aphanothece halophytica
synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine
S-adenosyl-L-homocysteine + N-methylglycine
-
-
?
S-adenosyl-L-methionine + N-methylglycine
Aphanothece halophytica
synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
?
S-adenosyl-L-methionine + sarcosine
Aphanothece halophytica
-
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Aphanothece halophytica
-
-
-
Aphanothece halophytica
Q83WC4
inducible enzyme
-
Purification (Commentary)
Commentary
Organism
; recombinant enzyme from Escherichia coli by ammonium sulfate fractionation and ion exchange chromatography to homogeneity
Aphanothece halophytica
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
the enzyme is involved in the alternate biosynthesis of betaine, which is an important osmoprotectant and is synthesized in response to abiotic stress
659286
Aphanothece halophytica
?
-
-
-
-
additional information
enzyme shows strict substrate specificity
659286
Aphanothece halophytica
?
-
-
-
-
S-adenosyl-L-methionine + glycine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + sarcosine
-
-
-
?
S-adenosyl-L-methionine + glycine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + sarcosine
i.e. 2-methylglycine
-
-
?
S-adenosyl-L-methionine + glycine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N-methylglycine
-
-
-
?
S-adenosyl-L-methionine + glycine
synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N-methylglycine
-
-
-
?
S-adenosyl-L-methionine + N-methylglycine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
-
?
S-adenosyl-L-methionine + N-methylglycine
synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
-
?
S-adenosyl-L-methionine + sarcosine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
-
?
Subunits
Subunits
Commentary
Organism
monomer
1 * 33000, recombinant enzyme, SDS-PAGE; 1 * 33000, SDS-PAGE, 1 * 32000, gel filtration
Aphanothece halophytica
More
enzyme aggregated to tetramers is inactive
Aphanothece halophytica
Temperature Optimum [°C]
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Aphanothece halophytica
pH Optimum
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.8
-
assay at
Aphanothece halophytica
9
-
glycine or N-methylglycine
Aphanothece halophytica
pH Range
pH Minimum
pH Maximum
Commentary
Organism
8
11
glycine or N-methylglycine, very low activity below pH 8
Aphanothece halophytica
Activating Compound (protein specific)
Activating Compound
Commentary
Organism
Structure
additional information
abiotic stess-inducible enzyme, e.g. high salt content
Aphanothece halophytica
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli BL21(DE3)
Aphanothece halophytica
orf1, DNA and amino acid sequence determination and analysis, phylogenetic analysis, overexpression of wild-type and mutant enzymes in Escherichia coli, functional coexpression of the soluble enzyme with the sarcosine dimethylglycine methyltransferase, EC 2.1.1.157, increases the intracellular betaine level and the growth rate of Escherichia coli strain BL21(DE3)
Aphanothece halophytica
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
R169E
site-directed mutagenesis, completely inactive mutant
Aphanothece halophytica
R169E
inactive
Aphanothece halophytica
R169K
site-directed mutagenesis, reduced activity and highly increased Km for ATP with both glycine and sarcosine compared to the wild-type enzyme
Aphanothece halophytica
R169K
drastic increase of Km of the methyl-acceptor substrates, 2-fold increase in Km for S-adenosyl-L-methionine
Aphanothece halophytica
Inhibitors (protein specific)
Inhibitors
Commentary
Organism
Structure
acetate
80% inhibition at 250 mM, with glycine
Aphanothece halophytica
acetate
-
Aphanothece halophytica
dimethylglycine
versus glycine, 65% inhibition at 250 mM
Aphanothece halophytica
ethanolamine
45% inhibition at 250 mM, with glycine
Aphanothece halophytica
ethanolamine
-
Aphanothece halophytica
KCl
-
Aphanothece halophytica
monomethyl ethanolamine
35% inhibition at 250 mM, with glycine
Aphanothece halophytica
monomethyl ethanolamine
-
Aphanothece halophytica
additional information
inhibition rates with sarcosine, overview, no feedback inhibition by betaine up to 2 M, no inhibition with glycine by serine, methionine, asparagine, glutamate, cysteine, and alanine
Aphanothece halophytica
additional information
no inhibition with betaine
Aphanothece halophytica
N,N-dimethylglycine
-
Aphanothece halophytica
NaCl
-
Aphanothece halophytica
phenylalanine
10% inhibition at 250 mM, with glycine
Aphanothece halophytica
proline
30% inhibition at 250 mM, with glycine
Aphanothece halophytica
proline
-
Aphanothece halophytica
S-adenosyl-L-homocysteine
product inhibition
Aphanothece halophytica
S-adenosyl-L-homocysteine
-
Aphanothece halophytica
sarcosine
competitive inhibition, versus glycine, 70% inhibition at 250 mM
Aphanothece halophytica
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.6
-
S-adenosyl-L-methionine
pH 8.8, 37°C, recombinant wild-type enzyme, with sarcosine
Aphanothece halophytica
0.6
-
S-adenosyl-L-methionine
with N-methylglycine as methyl group acceptor, pH 8.8, 37°C
Aphanothece halophytica
0.7
-
S-adenosyl-L-methionine
pH 8.8, 37°C, recombinant wild-type enzyme, with glycine
Aphanothece halophytica
0.7
-
S-adenosyl-L-methionine
with glycine as methyl group acceptor, pH 8.8, 37°C
Aphanothece halophytica
0.8
-
N-methylglycine
pH 8.8, 37°C
Aphanothece halophytica
0.8
-
sarcosine
pH 8.8, 37°C, recombinant wild-type enzyme
Aphanothece halophytica
1
-
glycine
pH 8.8, 37°C, recombinant mutant R169K
Aphanothece halophytica
1.1
-
sarcosine
pH 8.8, 37°C, recombinant mutant R169K
Aphanothece halophytica
1.5
-
glycine
pH 8.8, 37°C, recombinant wild-type enzyme
Aphanothece halophytica
1.5
-
glycine
pH 8.8, 37°C
Aphanothece halophytica
50
-
S-adenosyl-L-methionine
pH 8.8, 37°C, recombinant mutant R169K, with sarcosine
Aphanothece halophytica
75
-
S-adenosyl-L-methionine
pH 8.8, 37°C, recombinant mutant R169K, with glycine
Aphanothece halophytica
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
31210
-
calculated from the deduced amino acid sequence
Aphanothece halophytica
32000
-
recombinant enzyme, gel filtration
Aphanothece halophytica
32000
-
gel filtration
Aphanothece halophytica
33000
-
1 * 33000, recombinant enzyme, SDS-PAGE
Aphanothece halophytica
33000
-
1 * 33000, SDS-PAGE, 1 * 32000, gel filtration
Aphanothece halophytica
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
additional information
Aphanothece halophytica
the enzyme is involved in the alternate biosynthesis of betaine, which is an important osmoprotectant and is synthesized in response to abiotic stress
?
-
-
-
S-adenosyl-L-methionine + glycine
Aphanothece halophytica
-
S-adenosyl-L-homocysteine + sarcosine
-
-
?
S-adenosyl-L-methionine + glycine
Aphanothece halophytica
synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine
S-adenosyl-L-homocysteine + N-methylglycine
-
-
?
S-adenosyl-L-methionine + N-methylglycine
Aphanothece halophytica
synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
?
S-adenosyl-L-methionine + sarcosine
Aphanothece halophytica
-
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
-
Aphanothece halophytica
recombinant enzyme from Escherichia coli by ammonium sulfate fractionation and ion exchange chromatography to homogeneity
Aphanothece halophytica
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
additional information
the enzyme is involved in the alternate biosynthesis of betaine, which is an important osmoprotectant and is synthesized in response to abiotic stress
659286
Aphanothece halophytica
?
-
-
-
-
additional information
enzyme shows strict substrate specificity
659286
Aphanothece halophytica
?
-
-
-
-
S-adenosyl-L-methionine + glycine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + sarcosine
-
-
-
?
S-adenosyl-L-methionine + glycine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + sarcosine
i.e. 2-methylglycine
-
-
?
S-adenosyl-L-methionine + glycine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N-methylglycine
-
-
-
?
S-adenosyl-L-methionine + glycine
synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N-methylglycine
-
-
-
?
S-adenosyl-L-methionine + N-methylglycine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
-
?
S-adenosyl-L-methionine + N-methylglycine
synthesis of the compatible solute betaine de novo through the methylation of glycine, sarcosine and dimethylglycine with the methyl group from S-adenosylmethionine
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
-
?
S-adenosyl-L-methionine + sarcosine
-
659286
Aphanothece halophytica
S-adenosyl-L-homocysteine + N,N-dimethylglycine
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
monomer
1 * 33000, recombinant enzyme, SDS-PAGE
Aphanothece halophytica
monomer
1 * 33000, SDS-PAGE, 1 * 32000, gel filtration
Aphanothece halophytica
More
enzyme aggregated to tetramers is inactive
Aphanothece halophytica
Temperature Optimum [°C] (protein specific)
Temperature Optimum [°C]
Temperature Optimum Maximum [°C]
Commentary
Organism
37
-
assay at
Aphanothece halophytica
pH Optimum (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.8
-
assay at
Aphanothece halophytica
9
-
glycine or N-methylglycine
Aphanothece halophytica
pH Range (protein specific)
pH Minimum
pH Maximum
Commentary
Organism
8
11
glycine or N-methylglycine, very low activity below pH 8
Aphanothece halophytica
Other publictions for EC 2.1.1.156
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
733736
Niu
-
Co-expression of ApGSMT and Ap ...
Aphanothece halophytica
Environ. Exp. Bot.
104
16-25
2014
-
-
1
-
-
-
-
-
-
-
-
2
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
734932
Lai
Transgenic Arabidopsis express ...
Methanohalophilus portucalensis, Methanohalophilus portucalensis FDF-1
Plant Mol. Biol.
85
429-441
2014
-
-
1
-
-
-
-
-
-
-
-
4
-
5
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
-
-
1
1
-
-
-
734671
He
-
Co-expression of genes ApGSMT2 ...
Aphanothece halophytica
Mol. Breed.
31
559-573
2013
-
-
1
-
-
-
-
-
-
-
-
2
-
1
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
1
1
1
1
-
-
720836
Lai
Characterization and regulatio ...
Methanohalophilus portucalensis, Methanohalophilus portucalensis FDF1
PLoS ONE
6
e25090
2011
-
-
1
-
-
-
2
4
-
3
2
6
-
2
-
-
1
-
-
-
-
-
8
1
1
-
-
-
1
-
-
1
-
1
2
-
-
1
1
-
-
-
2
2
-
4
-
3
2
6
-
-
-
1
-
-
-
-
8
1
1
-
-
-
1
-
-
1
1
2
2
1
-
-
719715
Kimura
Glycine betaine biosynthesized ...
Myxococcus xanthus
J. Bacteriol.
192
1467-1470
2010
-
-
1
-
1
-
-
-
-
-
1
2
-
1
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
1
-
-
-
-
-
1
1
-
1
-
-
-
-
-
-
-
1
2
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
2
2
-
-
-
671647
Lu
Identification of glycine beta ...
Synechococcus sp. WH 8102
Arch. Microbiol.
186
495-506
2006
-
-
1
-
-
-
-
4
-
-
3
2
-
6
-
-
1
-
-
-
-
-
5
1
-
-
-
-
2
1
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
4
-
-
3
2
-
-
-
1
-
-
-
-
5
1
-
-
-
-
2
1
-
-
-
-
-
-
-
-
659286
Waditee
Isolation and functional chara ...
Aphanothece halophytica
J. Biol. Chem.
278
4932-4942
2003
1
-
1
-
2
-
12
9
-
-
3
5
-
2
-
-
1
-
-
-
-
-
9
2
1
-
-
-
2
1
-
-
-
-
-
1
-
2
-
-
4
-
-
18
-
12
-
-
5
5
-
-
-
2
-
-
-
-
9
3
1
-
-
-
2
1
-
-
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-
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2001
1
3
1
-
-
-
5
4
-
1
3
3
-
3
-
-
1
-
-
-
3
-
6
1
1
-
-
-
2
-
-
-
-
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1
3
1
-
-
-
-
-
5
-
4
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1
3
3
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1
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3
-
6
1
1
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-
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2
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659145
Nyyssola
Extreme halophiles synthesize ...
Actinopolyspora halophila, Halorhodospira halochloris
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2
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1
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1
6
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3
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1
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6
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13
1
2
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-
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2
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-
-
-
-
-
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2
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-
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1
-
-
-
-
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1
6
-
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1
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6
-
13
1
2
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2
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