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Literature summary extracted from

  • Usha, V.; Dover, L.G.; Roper, D.L.; Lloyd, A.J.; Besra, G.S.
    Use of a codon alteration strategy in a novel approach to cloning the Mycobacterium tuberculosis diaminopimelic acid epimerase (2006), FEMS Microbiol. Lett., 262, 39-47.
    View publication on PubMed

Application

EC Number Application Comment Organism
5.1.1.7 drug development the recombinant DapF produced is correctly folded and is a suitable tool for a drug development study Mycobacterium tuberculosis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.1.1.7 since previous attempts to express the diaminopimelate epimerase gene dapF of Mycobacterium tuberculosis in Escherichia coli results in undetectable enzyme yields a recombinant DapF protein is expressed in Escherichia coli consisting of silent mutation of the first 10 codons of the open reading frame in an attempt to reduce the formation of secondary structures that occur near the 5' end of the mRNA and inhibit translation. This significantly increases the yield of the enzyme Mycobacterium tuberculosis

Protein Variants

EC Number Protein Variants Comment Organism
5.1.1.7 additional information a recombinant DapF is generated consisting of silent mutation of the first 10 codons of the open reading frame. single nucleotide substitutions are incorporated without changing product composition in the first 30 nucleotides of the dapF open reading frame,in order to disrupt any secondary structure-promoting sequences present. this significantly increases the yield of the enzyme Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
5.1.1.7 12 17 meso-diaminoheptanedioate recombinant DapF consisting of silent mutation of the first 10 codons of the open reading frame Mycobacterium tuberculosis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
5.1.1.7 30000
-
 molecular weight of recombinant DapF consisting of silent mutation of the first 10 codons, determined by SDS-PAGE Mycobacterium tuberculosis

Organism

EC Number Organism UniProt Comment Textmining
5.1.1.7 Mycobacterium tuberculosis
-
 H37Rv
-
5.1.1.7 Mycobacterium tuberculosis H37Rv
-
 H37Rv
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.1.1.7 protein is applied to a Ni21-primed chelating sepharose column, and DapF-containing eluate, fractions are dialysed and further purified by anion exchange chromatography on a HiTrap Q Sepharose FF column, representing a yield of 1 mg/L culture Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.1.1.7 LL-2,6-Diaminoheptanedioate
-
 Mycobacterium tuberculosis meso-Diaminoheptanedioate
-
 ?
5.1.1.7 LL-2,6-Diaminoheptanedioate
-
 Mycobacterium tuberculosis H37Rv meso-Diaminoheptanedioate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
5.1.1.7 DapF
-
 Mycobacterium tuberculosis
5.1.1.7 Diaminopimelic acid epimerase
-
 Mycobacterium tuberculosis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
5.1.1.7 30
-
 recombinant DapF consisting of silent mutation of the first 10 codons of the open reading frame is almost 50% more active at 30°C than at 25°C Mycobacterium tuberculosis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
5.1.1.7 7.5
-
 maximum activity of recombinant DapF consisting of silent mutation of the first 10 codons of the open reading frame Mycobacterium tuberculosis

pH Range

EC Number pH Minimum pH Maximum Comment Organism
5.1.1.7 6.5 9 recombinant DapF consisting of silent mutation of the first 10 codons of the open reading frame Mycobacterium tuberculosis