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Literature summary extracted from

  • Annamalai, T.; Dani, N.; Cheng, B.; Tse-Dinh, Y.C.
    Analysis of DNA relaxation and cleavage activities of recombinant Mycobacterium tuberculosis DNA topoisomerase I from a new expression and purification protocol (2009), BMC Biochem., 10, 18.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
5.6.2.1 drug development the enzyme is an attractive target for discovery of novel tuberculosis drugs that act by enhancing the accumulation of the topoisomerase-DNA cleavage product Mycobacterium tuberculosis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.6.2.1 expression of the soluble His6-tagged enzyme in Escherichia coli strain BL21 (DE3) Mycobacterium tuberculosis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
5.6.2.1 Mg2+ required for DNA substrate relaxation, optimal at 2.5-7.5 mM Escherichia coli
5.6.2.1 Mg2+ required for DNA substrate relaxation, optimal at 5-12.5 mM Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
5.6.2.1 Escherichia coli
-
-
-
5.6.2.1 Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.6.2.1 His6-tagged recombinant enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography, cleavage and elimination of the His-tag Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.6.2.1 additional information mapping cleavage sites on single-stranded DNA substrate Mtop with length of 216 bases amplified from topA gene of Mycobacterium tuberculosis, substrate is negatively supercoiled DNA Mycobacterium tuberculosis ?
-
 ?
5.6.2.1 additional information substrate is negatively supercoiled DNA Escherichia coli ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
5.6.2.1 DNA topoisomerase I
-
 Escherichia coli
5.6.2.1 DNA topoisomerase I
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.6.2.1 37
-
 DNA relaxation assay at Escherichia coli
5.6.2.1 37
-
 DNA relaxation assay at Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.6.2.1 8
-
 DNA relaxation assay at Escherichia coli
5.6.2.1 8
-
 DNA relaxation assay at Mycobacterium tuberculosis

Cofactor

EC Number Cofactor Comment Organism Structure
5.6.2.1 additional information no ATP required Mycobacterium tuberculosis