Application | Comment | Organism |
---|---|---|
medicine | the enzyme is involved in biosynthesis of D-mycaminose. D-Mycaminose is an unusual dideoxy sugar found attached to the antibiotic tylosin, a commonly used veterinarian therapeutic | Streptomyces fradiae |
Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Streptomyces fradiae |
Crystallization (Comment) | Organism |
---|---|
hanging drop method of vapor diffusion method, high resolution X-ray structures of TylM1, one in which the enzyme contains bound SAM and dTDP-phenol and the second in which the protein is complexed with S-adenosyl-L-homocysteine and dTDP-3-amino-3,6-dideoxyglucose, its natural substrate. Combined, these two structures, solved to 1.35 A and 1.79 A resolution, respectively, show the orientations of SAM and the dTDP-linked sugar substrate within the active site region. Specifically, the C-30 amino group of the hexose is in the correct position for an in-line attack at the reactive methyl group of S-adenosyl-L-methionine. High-resolution X-ray models show that the observed perturbations in the kinetic constants of the mutant enzynes H123A and H123N are not due to major changes in their threedimensional folds. Most likely the proton on the C-3' amino group is transferred to one of the water molecules lining the active site pocket as catalysis proceeds | Streptomyces fradiae |
Protein Variants | Comment | Organism |
---|---|---|
H123A | mutant protein retains catalytic activity, the kcat/Km is reduced to 1.8% compared to the wild-type enzyme | Streptomyces fradiae |
H123N | mutant protein retains catalytic activity, the kcat/Km is reduced to 0.37% compared to the wild-type enzyme | Streptomyces fradiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.045 | - |
dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | pH 8.5, 37°C, wild-type enzyme | Streptomyces fradiae | |
0.67 | - |
dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | pH 8.5, 37°C, mutant enzyme H123A | Streptomyces fradiae | |
0.93 | - |
dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | pH 8.5, 37°C, mutant enzyme H123N | Streptomyces fradiae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 S-adenosyl-L-methionine + dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | Streptomyces fradiae | the enzyme is involved in biosynthesis of D-mycaminose. It is synthesized by the Gram-positive bacterium Streptomyces fradiae as a dTDP-linked sugar | 2 S-adenosyl-L-homocysteine + dTDP-3-dimethylamino-3,6-dideoxy-alpha-D-glucopyranose | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces fradiae | P95748 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Streptomyces fradiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 S-adenosyl-L-methionine + dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | - |
Streptomyces fradiae | 2 S-adenosyl-L-homocysteine + dTDP-3-dimethylamino-3,6-dideoxy-alpha-D-glucopyranose | - |
? | |
2 S-adenosyl-L-methionine + dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | the enzyme is involved in biosynthesis of D-mycaminose. It is synthesized by the Gram-positive bacterium Streptomyces fradiae as a dTDP-linked sugar | Streptomyces fradiae | 2 S-adenosyl-L-homocysteine + dTDP-3-dimethylamino-3,6-dideoxy-alpha-D-glucopyranose | - |
? |
Synonyms | Comment | Organism |
---|---|---|
TylM1 | - |
Streptomyces fradiae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Streptomyces fradiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0126 | - |
dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | pH 8.5, 37°C, mutant enzyme H123N | Streptomyces fradiae | |
0.046 | - |
dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | pH 8.5, 37°C, mutant enzyme H123A | Streptomyces fradiae | |
0.172 | - |
dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | pH 8.5, 37°C, wild-type enzyme | Streptomyces fradiae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Streptomyces fradiae |
General Information | Comment | Organism |
---|---|---|
physiological function | the enzyme is involved in biosynthesis of D-mycaminose. It is synthesized by the Gram-positive bacterium Streptomyces fradiae as a dTDP-linked sugar | Streptomyces fradiae |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0135 | - |
dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | pH 8.5, 37°C, mutant enzyme H123N | Streptomyces fradiae | |
0.069 | - |
dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | pH 8.5, 37°C, mutant enzyme H123A | Streptomyces fradiae | |
3.8 | - |
dTDP-3-amino-3,6-dideoxy-alpha-D-glucopyranose | pH 8.5, 37°C, wild-type enzyme | Streptomyces fradiae |