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Literature summary for 2.1.1.343 extracted from

  • Jankowitsch, F.; Kühm, C.; Kellner, R.; Kalinowski, J.; Pelzer, S.; Macheroux, P.; Mack, M.
    A novel N,N-8-amino-8-demethyl-D-riboflavin dimethyltransferase (RosA) catalyzing the two terminal steps of roseoflavin biosynthesis in Streptomyces davawensis (2011), J. Biol. Chem., 286, 38275-285.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Rosetta 2 (DE3), recombinant His6-tagged RosA purified, ORF sda77220 is named rosA is located in the middle of a putative gene cluster within the genome of Streptomyces davawensis Streptomyces davaonensis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0286
-
S-adenosyl-L-methionine pH 8.0, 37°C Streptomyces davaonensis
0.0577
-
8-amino-8-demethyl-D-riboflavin pH 8.0, 37°C Streptomyces davaonensis

Localization

Localization Comment Organism GeneOntology No. Textmining
additional information cell-free extract, roseoflavin synthesis is observed in the exponentially phase growth, under laboratory conditions using starch and yeast extract containing broth, roseoflavin synthesis is observed in the stationary phase of growth Streptomyces davaonensis
-
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
37940
-
calculated Streptomyces davaonensis
38000
-
1 * 38000, calculated, purified recombinant RosA Streptomyces davaonensis
40000
-
SDS-PAGE Streptomyces davaonensis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
8-amino-8-demethyl-D-riboflavin + S-adenosyl-L-methionine Streptomyces davaonensis
-
roseoflavin + S-adenosyl-L-homocysteine the products isolated by HPLC and identified by mass spectrometry ir
8-amino-8-demethyl-D-riboflavin + S-adenosyl-L-methionine Streptomyces davaonensis 768
-
roseoflavin + S-adenosyl-L-homocysteine the products isolated by HPLC and identified by mass spectrometry ir

Organism

Organism UniProt Comment Textmining
Streptomyces davaonensis E7BBN5
-
-
Streptomyces davaonensis 768 E7BBN5
-
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate precipitation, phenyl-Sepharose chromatography, anion exchange chromatography, gel filtration on a HiLoad 16/60 Superdex 200 column, analyzed using SDS-PAGE and MS/MS Streptomyces davaonensis

Reaction

Reaction Comment Organism Reaction ID
2 S-adenosyl-L-methionine + 8-amino-8-demethylriboflavin = 2 S-adenosyl-L-homocysteine + roseoflavin roseoflavin, i.e. 1-deoxy-1-[8-(dimethylamino)-7-methyl-2,4-dioxo-3,4-dihydrobenzo[g]pteridin-10(2H)-yl]pentitol, a two-step methylation reaction Streptomyces davaonensis
S-adenosyl-L-methionine + 8-amino-8-demethylriboflavin = S-adenosyl-L-homocysteine + 8-demethyl-8-(methylamino)riboflavin 1a Streptomyces davaonensis
S-adenosyl-L-methionine + 8-demethyl-8-(methylamino)riboflavin = S-adenosyl-L-homocysteine + roseoflavin 1b Streptomyces davaonensis

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.58
-
pH 8.0, 37°C Streptomyces davaonensis

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
8-amino-8-demethyl-D-riboflavin + S-adenosyl-L-methionine
-
Streptomyces davaonensis roseoflavin + S-adenosyl-L-homocysteine the products isolated by HPLC and identified by mass spectrometry ir
8-amino-8-demethyl-D-riboflavin + S-adenosyl-L-methionine
-
Streptomyces davaonensis roseoflavin + S-adenosyl-L-homocysteine products isolated by HPLC and identified by mass spectrometry ir
8-amino-8-demethyl-D-riboflavin + S-adenosyl-L-methionine
-
Streptomyces davaonensis 768 roseoflavin + S-adenosyl-L-homocysteine the products isolated by HPLC and identified by mass spectrometry ir
8-amino-8-demethyl-D-riboflavin + S-adenosyl-L-methionine
-
Streptomyces davaonensis 768 roseoflavin + S-adenosyl-L-homocysteine products isolated by HPLC and identified by mass spectrometry ir
additional information no activity with 8-hydroxy-8-demethyl-D-riboflavin Streptomyces davaonensis ?
-
?
additional information no activity with 8-hydroxy-8-demethyl-D-riboflavin Streptomyces davaonensis 768 ?
-
?

Subunits

Subunits Comment Organism
monomer 1 * 38000, calculated, purified recombinant RosA Streptomyces davaonensis

Synonyms

Synonyms Comment Organism
N,N-8-amino-8-demethyl-D-riboflavin dimethyltransferase
-
Streptomyces davaonensis
RosA rosA: gene name Streptomyces davaonensis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Streptomyces davaonensis
52
-
-
Streptomyces davaonensis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.24
-
S-adenosyl-L-methionine pH 8.0, 37°C Streptomyces davaonensis
0.37
-
8-amino-8-demethyl-D-riboflavin pH 8.0, 37°C Streptomyces davaonensis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Streptomyces davaonensis
8.8
-
-
Streptomyces davaonensis

Cofactor

Cofactor Comment Organism Structure
S-adenosyl-L-methionine
-
Streptomyces davaonensis

General Information

General Information Comment Organism
physiological function RosA catalyzes the two terminal steps in the biosynthesis of the antibiotic roseoflavin. RosA is the first enzyme of the biosynthesis to be identified Streptomyces davaonensis