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 (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 | 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 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 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 | UniProt | Comment | Textmining |
---|---|---|---|
Mycobacterium tuberculosis | - |
H37Rv | - |
Mycobacterium tuberculosis H37Rv | - |
H37Rv | - |
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 | 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 | Comment | Organism |
---|---|---|
DapF | - |
Mycobacterium tuberculosis |
Diaminopimelic acid epimerase | - |
Mycobacterium tuberculosis |
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 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 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 |