Application | Comment | Organism |
---|---|---|
synthesis | the enzyme is used for the preparation of flavin mononucleotide (FMN) and FMN analogues from their corresponding riboflavin precursors, which is performed in a two-step procedure. After initial enzymatic conversion of riboflavin to FAD by the bifunctional FAD synthetase, the adenyl moiety of FAD is hydrolyzed with snake venom phosphodiesterase to yield FMN. The engineered FAD synthetase from Corynebacterium ammoniagenes with deleted N-terminal adenylation domain is a biocatalyst that is stable and efficient for direct and quantitative phosphorylation of riboflavin and riboflavin analogues to their corresponding FMN cofactors at preparative-scale | Corynebacterium ammoniagenes |
Cloned (Comment) | Organism |
---|---|
recombinant expression of C-terminally poly-His-tagged N-terminally truncated mutant in Escherichia coli | Corynebacterium ammoniagenes |
Protein Variants | Comment | Organism |
---|---|---|
additional information | engineering of the FAD synthetase from Corynebacterium ammoniagenes by deleting its N-terminal adenylation domain leads to a biocatalyst that is stable and efficient for direct and quantitative phosphorylation of riboflavin and riboflavin analogues to their corresponding FMN cofactors at preparative-scale. Deletion of the N-terminal adenosyl transfer domain in the truncated C-terminal RF kinase domain, tcRFK, variants results in a drop in the TM value from 40°C (parental CaFADS) to 35°C for tcRFK. Addition of the C-terminal poly-His tag further reduces the TM to 30°C, presumably due to the conformationally flexible tail formed by the extra amino acids | Corynebacterium ammoniagenes |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Corynebacterium ammoniagenes |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
additional information | - |
the elution profile of full-length FADS shows two characteristic peaks at molecular weights corresponding to its monomeric and trimeric forms, while the tcRFK elutes as a single peak at an estimated molecular weight of 16.6 kDa, consistent with monomeric enzyme | Corynebacterium ammoniagenes |
16600 | - |
recombinant truncated C-terminal RF kinase domain, gel filtration | Corynebacterium ammoniagenes |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + FMN | Corynebacterium ammoniagenes | engineered mutant | diphosphate + FAD | - |
? | |
additional information | Corynebacterium ammoniagenes | bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26 | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Corynebacterium ammoniagenes | Q59263 | gene ribF | - |
Purification (Comment) | Organism |
---|---|
recombinant C-terminally poly-His-tagged N-terminally truncated mutant_187-338 from Escherichia coli by ion exchange chromatography and gel filtration | Corynebacterium ammoniagenes |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + FMN | engineered mutant | Corynebacterium ammoniagenes | diphosphate + FAD | - |
? | |
additional information | bifunctional FAD synthetase exhibiting both the activities of FAD synthetase, EC 2.7.7.2, and riboflavin kinase, EC 2.7.1.26 | Corynebacterium ammoniagenes | ? | - |
? | |
additional information | the engineered FAD synthetase from Corynebacterium ammoniagenes with deleted N-terminal adenylation domain is a biocatalyst that is stable and efficient for direct and quantitative phosphorylation of riboflavin and riboflavin analogues to their corresponding FMN cofactors at preparative-scale, method evaluation, overview | Corynebacterium ammoniagenes | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
monomer | x * 16600, recombinant truncated C-terminal RF kinase domain, SDS-PAGE | Corynebacterium ammoniagenes |
Synonyms | Comment | Organism |
---|---|---|
FADS | - |
Corynebacterium ammoniagenes |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Corynebacterium ammoniagenes |