Application | Comment | Organism |
---|---|---|
synthesis | biosynthesis of tetracycline from anhydrotetracycline in Saccharomyces cerevisiae heterologously expressing the anhydrotetracycline hydroxylase OxyS, the dehydrotetracycline reductase CtcM, and the F420 reductase FNO from three bacterial hosts. This biosynthesis of tetracycline is enabled by OxyS performing just one hydroxylation step in S. cerevisiae | Streptomyces rimosus subsp. rimosus |
Cloned (Comment) | Organism |
---|---|
gene oxyS, functional recombinant overexpression of C-terminally FLAG-tagged enzyme in Saccharomyces cerevisiae from plasmid pSP-G1 resulting in biosynthesis of dehydrotetracycline from anhydrotetracycline, functional coexpression with heterologous dehydrotetracycline reductase CtcM, and the F420 reductase FNO | Streptomyces sp. |
Protein Variants | Comment | Organism |
---|---|---|
additional information | recombinant overexpression of gene oxySin Saccharomyces cerevisiae, functional coexpression with heterologous dehydrotetracycline reductase CtcM, and the F420 reductase FNO. Biosynthesis of tetracycline is enabled by OxyS performing just one hydroxylation step in Saccharomyces cerevisiae despite its previous characterization as a double hydroxylase. This single hydroxylation enables the purification and structural characterization of a hypothetical intermediate in oxytetracycline biosynthesis that can explain structural differences between oxytetracycline and chlortetracycline. Using the alternative enzyme CtcM from the chlortetracycline pathway instead of OxyR yields in vitro an increased ratio of tetracycline to oxytetracycline. A unique cofactor to the last steps of the tetracyclines' biosynthesis that is not native to Saccharomyces cerevisiae is cofactor F420, a lactyl oligoglutamate phosphodiester derivative of 7,8-didemethyl-8-hydroxy-5-deazariboflavin (Fo). Fo is much more synthetically accessible than F420. Fo can replace F420 in tetracycline biosynthesis. Three F420 reductase candidates from Mycobacterium tuberculosis, Archaeoglobus fulgidus, and Streptomyces griseus are explored, and the enzyme from Archaeoglobus fulgidus is chosen. Method development and evaluation, method optimization, overview | Streptomyces sp. |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
anhydrotetracycline + NADPH + H+ + O2 | Streptomyces sp. | - |
12-dehydrotetracycline + NADP+ + H2O | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces rimosus subsp. rimosus | L8EUQ6 | cf. EC 1.14.13.234 | - |
Streptomyces rimosus subsp. rimosus DSM 40260 | L8EUQ6 | cf. EC 1.14.13.234 | - |
Streptomyces sp. | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
anhydrotetracycline + NADPH + H+ + O2 | - |
Streptomyces sp. | 12-dehydrotetracycline + NADP+ + H2O | - |
? | |
anhydrotetracycline + NADPH + H+ + O2 | - |
Streptomyces rimosus subsp. rimosus | 5a,11a-dehydrotetracycline + NADP+ + H2O | - |
? | |
anhydrotetracycline + NADPH + H+ + O2 | - |
Streptomyces rimosus subsp. rimosus DSM 40260 | 5a,11a-dehydrotetracycline + NADP+ + H2O | - |
? |
Synonyms | Comment | Organism |
---|---|---|
anhydrotetracycline hydroxylase | - |
Streptomyces sp. |
oxyS | - |
Streptomyces sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NADPH | - |
Streptomyces sp. |