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Literature summary extracted from

  • Kudo, F.; Fujii, T.; Kinoshita, S.; Eguchi, T.
    Unique O-ribosylation in the biosynthesis of butirosin (2007), Bioorg. Med. Chem., 15, 4360-4368.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.2.49 gene btrL, soluble BtrL is only obtained by co-expression with the GroESL chaperones8 at low temperature in Escherichia coli BL21(DE3) Niallia circulans
3.1.3.88 expressed in Escherichia coli BL21(DE3) cells Niallia circulans

General Stability

EC Number General Stability Organism
2.4.2.49 the recombinant BtrL protein is largely unstable and loses activity within hours, addition of 10% glycerol in assay mixtures Niallia circulans

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.4.2.49 Cu2+ high inhibition at 1 mM Niallia circulans
2.4.2.49 EDTA complete inhibition at 1 mM Niallia circulans
2.4.2.49 Mn2+
-
 Niallia circulans
2.4.2.49 Zn2+ high inhibition at 1 mM Niallia circulans
3.1.3.88 additional information EDTA does not affect the enzyme activity Niallia circulans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.4.2.49 Co2+ activates Niallia circulans
2.4.2.49 Mg2+ activates Niallia circulans
2.4.2.49 additional information BtrL requires essentially a divalent metal ion, optimally Mg2+, Ni2+, or Co2+ Niallia circulans
2.4.2.49 Ni2+ activates Niallia circulans
3.1.3.88 additional information the enzyme does not need a (divalent) metal ion for activity Niallia circulans

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.2.49 neamine + 5-phospho-alpha-D-ribose 1-diphosphate Niallia circulans
-
 5''-phosphoribostamycin + diphosphate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.2.49 Niallia circulans
-
-
-
2.4.2.49 Niallia circulans
-
 gene btrL
-
2.4.2.49 Niallia circulans SANK72073
-
-
-
2.4.2.49 Niallia circulans SANK72073
-
 gene btrL
-
3.1.3.88 Niallia circulans
-
-
-
3.1.3.88 Niallia circulans SANK72073
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.3.88 DEAE-Sephadex gel filtration and Superdex 200 gel filtration Niallia circulans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.2.49 additional information Btr enzymes substrate specificities, mass spectrometric analysis, overview Niallia circulans ?
-
 ?
2.4.2.49 additional information Btr enzymes substrate specificities, mass spectrometric analysis, overview Niallia circulans SANK72073 ?
-
 ?
2.4.2.49 neamine + 5-phospho-alpha-D-ribose 1-diphosphate
-
 Niallia circulans 5''-phosphoribostamycin + diphosphate
-
 ?
3.1.3.88 4-nitrophenyl phosphate + H2O
-
 Niallia circulans 4-nitrophenol + phosphate
-
 ?
3.1.3.88 4-nitrophenyl phosphate + H2O
-
 Niallia circulans SANK72073 4-nitrophenol + phosphate
-
 ?
3.1.3.88 5''-phosphoribostamycin + H2O
-
 Niallia circulans ribostamycin + phosphate
-
 ?
3.1.3.88 5''-phosphoribostamycin + H2O
-
 Niallia circulans SANK72073 ribostamycin + phosphate
-
 ?
3.1.3.88 D-fructose-1,6-bisphosphate + H2O rapid hydrolysis Niallia circulans ?
-
 ?
3.1.3.88 D-fructose-1,6-bisphosphate + H2O rapid hydrolysis Niallia circulans SANK72073 ?
-
 ?
3.1.3.88 additional information PRPP, fructose-6-phosphate, ribose-5-phosphate, and nucleotides (AMP, CMP, GMP, UMP, and dTMP) are not hydrolyzed Niallia circulans ?
-
 ?
3.1.3.88 additional information PRPP, fructose-6-phosphate, ribose-5-phosphate, and nucleotides (AMP, CMP, GMP, UMP, and dTMP) are not hydrolyzed Niallia circulans SANK72073 ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.2.49 btrL
-
 Niallia circulans
2.4.2.49 neoM
-
 Niallia circulans
3.1.3.88 btrP
-
 Niallia circulans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.2.49 27 32
-
 Niallia circulans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.2.49 7.5
-
-
 Niallia circulans

General Information

EC Number General Information Comment Organism
2.4.2.49 metabolism BtrA, BtrL, BtrP, and BtrV proteins encoded in the butirosin biosynthetic gene cluster (btr) from Bacillus circulans SANK72073 are responsible for the O-ribosylation process leading to the formation of ribostamycin, a key intermediate in this, and related antibiotic biosynthetic pathways. Both BtrL and BtrP are responsible for the ribosylation of neamine, using 5-phosphoribosyl 1-diphosphate as the ribosyl donor via two discrete steps: with BtrL first catalyzes the phosphoribosylaion of neamine to form 5''-phosphoribostamycin, followed by a BtrP catalyzing the dephosphorylation to generate ribostamycin Niallia circulans
2.4.2.49 additional information PRPP binding motif in BtrL Niallia circulans
3.1.3.88 metabolism the enzyme is responsible for the formation of the key ribostamycin intermediate in the biosynthesis of butirosin Niallia circulans