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Literature summary for 5.6.2.1 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
Search for more literature for 5.6.2.1

Application

Application Comment Organism
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)

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

Metals/Ions

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

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Mycobacterium tuberculosis
-
-
-

Purification (Commentary)

Purification (Comment) Organism
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)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
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 ?
-
 ?
additional information substrate is negatively supercoiled DNA Escherichia coli ?
-
 ?

Synonyms

Synonyms Comment Organism
DNA topoisomerase I
-
 Escherichia coli
DNA topoisomerase I
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

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

pH Optimum

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

Cofactor

Cofactor Comment Organism Structure
additional information no ATP required Mycobacterium tuberculosis