Literature summary for 5.1.1.7 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.

Application

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

Cloned(Commentary)

Cloned (Comment)	Organism
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

Cloned (Comment)

Organism

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

Protein Variants	Comment	Organism
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]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
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]

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

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	-	H37Rv	-
Mycobacterium tuberculosis H37Rv	-	H37Rv	-

Purification (Commentary)

Purification (Comment)	Organism
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)

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

Synonyms

Synonyms	Comment	Organism
DapF	-	Mycobacterium tuberculosis
Diaminopimelic acid epimerase	-	Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
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

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
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

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