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Literature summary for 4.1.99.14 extracted from

  • Silver, S.C.; Chandra, T.; Zilinskas, E.; Ghose, S.; Broderick, W.E.; Broderick, J.B.
    Complete stereospecific repair of a synthetic dinucleotide spore photoproduct by spore photoproduct lyase (2010), J. Biol. Inorg. Chem., 15, 943-955.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.1.99.14

Activating Compound

Activating Compound Comment Organism Structure
DTT
-
 Clostridium acetobutylicum

Cloned(Commentary)

Cloned (Comment) Organism
gene splB, functional expression of His6-tagged enzyme in Escherichia coli Tuner(DE3)pLysS Clostridium acetobutylicum
heterologously expressed in Escherichia coli as a His-tagged fusion protein Clostridium acetobutylicum

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ the purified enzyme contains between 2.3 and 3.1 iron atoms per protein molecule. A [3Fe-4S]+ cluster accounts for 3-4% of the iron, and a [4Fe-4S]+ cluster accounts for 34-45% of total iron. The SP lyase can be photoreduced under anaerobic conditions in the presence of DTT, tris(hydroxymethyl)aminomethane, and 5-deazariboflavin Clostridium acetobutylicum
iron-sulfur centre the UV-vis spectrum of the purified enzyme is characteristic of the presence of an iron-sulfur cluster. Purified enzyme contains between 2.3 and 3.1 iron atoms per protein. Electron paramagnetic resonance (EPR) spectroscopy reveals a [3Fe-4S]+ cluster accounting for 3-4% of the iron in the sample. Upon reduction, a signal characteristic of a [4Fe-4S]+ cluster is observed that accounts for 34-45% of total iron in the sample Clostridium acetobutylicum

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
41000
-
 SDS-PAGE Clostridium acetobutylicum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) + S-adenosyl-L-methionine + 2 H+ Clostridium acetobutylicum
-
 thymidylyl-(3'-5')-thymidylate (in DNA) + 5'-deoxyadenosine + L-methionine
-
 ?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) + S-adenosyl-L-methionine + 2 H+ Clostridium acetobutylicum ATCC 824D
-
 thymidylyl-(3'-5')-thymidylate (in DNA) + 5'-deoxyadenosine + L-methionine
-
 ?

Organism

Organism UniProt Comment Textmining
Clostridium acetobutylicum
-
-
-
Clostridium acetobutylicum
-
 gene splB
-
Clostridium acetobutylicum ATCC 824D
-
 gene splB
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli Tuner(DE3)pLysS by nickel affinity chromatography to homogeneity Clostridium acetobutylicum
using a HisTrap HP 5-mL column Clostridium acetobutylicum

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0071
-
 pH 7.5, 30°C Clostridium acetobutylicum
0.0071
-
 reduced purified recombinant enzyme, pH 7.5, temperature not specified in the publication Clostridium acetobutylicum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) + S-adenosyl-L-methionine + 2 H+
-
 Clostridium acetobutylicum thymidylyl-(3'-5')-thymidylate (in DNA) + 5'-deoxyadenosine + L-methionine
-
 ?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) + S-adenosyl-L-methionine + 2 H+ the reduced enzyme rapidly and completely repairs the 5R-diastereomer of a synthetic dinucleotide SP, whereas no repair occurs with the 5S-diastereomer Clostridium acetobutylicum thymidylyl-(3'-5')-thymidylate (in DNA) + 5'-deoxyadenosine + L-methionine
-
 ?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) + S-adenosyl-L-methionine + 2 H+
-
 Clostridium acetobutylicum ATCC 824D thymidylyl-(3'-5')-thymidylate (in DNA) + 5'-deoxyadenosine + L-methionine
-
 ?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double helical DNA) + S-adenosyl-L-methionine + 2 H+ the reduced enzyme rapidly and completely repairs the 5R-diastereomer of a synthetic dinucleotide SP, whereas no repair occurs with the 5S-diastereomer Clostridium acetobutylicum ATCC 824D thymidylyl-(3'-5')-thymidylate (in DNA) + 5'-deoxyadenosine + L-methionine
-
 ?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine + S-adenosyl-L-methionine no repair is observed for the (5S) diasteroisomer Clostridium acetobutylicum thymidylyl-(3'->5')-thymidylate + 5'-deoxyadenosine + L-methionine
-
 ?
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine + S-adenosyl-L-methionine no repair is observed for the (5S) diasteroisomer Clostridium acetobutylicum ATCC 824D thymidylyl-(3'->5')-thymidylate + 5'-deoxyadenosine + L-methionine
-
 ?

Subunits

Subunits Comment Organism
? x * 41000, recombinant His-tagged enzyme, SDS-PAGE Clostridium acetobutylicum

Synonyms

Synonyms Comment Organism
More SP lyase is a member of the radical S-adenosyl-L-methionine superfamily of enzymes Clostridium acetobutylicum
SP lyase
-
 Clostridium acetobutylicum
spore photoproduct lyase
-
 Clostridium acetobutylicum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
 assay at Clostridium acetobutylicum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Clostridium acetobutylicum

Cofactor

Cofactor Comment Organism Structure
S-adenosyl-L-methionine
-
 Clostridium acetobutylicum

General Information

General Information Comment Organism
additional information spectral analysis of the purified recombinant enzyme, overview Clostridium acetobutylicum