Cloned (Comment) | Organism |
---|---|
gene splB, overexpression of His6-tagged wild-type and mutant enzymes and co-expression with plasmid pDB1282, in Escherichia coli strain BL21(DE3) | Bacillus subtilis |
Protein Variants | Comment | Organism |
---|---|---|
C141A | site-directed mutagenesis, the mutant repairs spore photoproduct at a rate which is about 3fold slower than the wild-type enzyme, the mutant shows altered competitive kinetic isotope effects compared to the wild-type enzyme. S-adenosyl-L-methionine plays a non-catalytical role in the mutant in contrast to the wild-type enzyme | Bacillus subtilis |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Dithionite | - |
Bacillus subtilis | |
DTT | - |
Bacillus subtilis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetics and competitive kinetic isotope effects of recombinant wild-type and mutant enzymes, overview | Bacillus subtilis |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | metalloenzyme, the enzyme uses a [4Fe-4S]1+ cluster to reduce the S-adenosyl-L-methionine generating a catalytic 5'-deoxyadenosyl radical | Bacillus subtilis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | Bacillus subtilis | - |
thymidylyl-(3'->5')-thymidylate | - |
? | |
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | Bacillus subtilis 168 | - |
thymidylyl-(3'->5')-thymidylate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Bacillus subtilis | - |
gene splB | - |
Bacillus subtilis 168 | - |
gene splB | - |
Purification (Comment) | Organism |
---|---|
recombinant His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity and cation exchange chromatography | Bacillus subtilis |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine (in double-helical DNA) = thymidylyl-(3'->5')-thymidylate (in double-helical DNA) | reaction mechanism, overview | Bacillus subtilis |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | - |
Bacillus subtilis | thymidylyl-(3'->5')-thymidylate | - |
? | |
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | the reaction is highly stereo-selective | Bacillus subtilis | thymidylyl-(3'->5')-thymidylate | - |
? | |
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | - |
Bacillus subtilis 168 | thymidylyl-(3'->5')-thymidylate | - |
? | |
(5R)-5,6-dihydro-5-(thymidin-7-yl)thymidine | the reaction is highly stereo-selective | Bacillus subtilis 168 | thymidylyl-(3'->5')-thymidylate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
SPL | - |
Bacillus subtilis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
22 | - |
assay at room temperature | Bacillus subtilis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Bacillus subtilis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
S-adenosyl-L-methionine | a radical S-adenosyl-L-methionine enzyme that uses a [4Fe-4S]1+ cluster to reduce the S-adenosyl-L-methionine generating a catalytic 5'-deoxyadenosyl radical | Bacillus subtilis |
Organism | Comment | pI Value Maximum | pI Value |
---|---|---|---|
Bacillus subtilis | recombinant His6-tagged mutant C141A, isoelectric focussing | - |
8.5 |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme is a member of the radical SAM superfamily | Bacillus subtilis |
malfunction | the enzyme C141A mutant produces thymidylyl-(3'->5')-thymidylate likely via an unprecedented thymine radical cation reduction (proton coupled electron transfer) mechanism | Bacillus subtilis |
additional information | residue C141 is solvent exposable and no other protein residue locates between the thymidylyl-(3'->5')-thymidylate radical and the C141 in the wild--type enzyme reaction pathway | Bacillus subtilis |
physiological function | spore photoproduct lyase repairs a covalent UV-induced thymine dimer, spore photoproduct, in germinating endospores and is responsible for endospores' strong UV resistance. The spore photoproduct is rapidly repaired by the metalloenzyme spore photoproduct lyase when spores start germinating | Bacillus subtilis |