Application | Comment | Organism |
---|---|---|
medicine | the requirement for an additional protein component, DesVIII, for activity must be taken into consideration in the design of combinatorial biosynthetic experiments with new glycosylated macrolides | Streptomyces venezuelae |
Cloned (Comment) | Organism |
---|---|
expression of desVII asaC-terminally His6-tagged protein in Escherichia coli. When affinity-tagged DesVII and DesVIII proteins are coproduced in Escherichia coli, they form a tight (alphabeta)3 complex that is at least 1000-fold more active than DesVII alone. The formation of the DesVII/DesVIII complex requires coexpression of both genes in vivo and cannot be fully achieved by mixing the individual protein components in vitro | Streptomyces venezuelae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.06 | - |
10-deoxymethynolide | pH 8.0, 29°C, addition of the DesIII protein, cosubstrate: 1.0 mM dTDP-3-amino-4,6-dideoxy-alpha-D-glucopyranose | Streptomyces venezuelae | |
0.16 | - |
TDP-3-dimethylamino-4,6-dideoxy-alpha-D-glucopyranose | pH 8.0, 29°C, addition of the DesIII protein, cosubstrate: 9.1 mM 10-deoxymethynolide | Streptomyces venezuelae | |
1.84 | - |
TDP-3-dimethylamino-4,6-dideoxy-alpha-D-glucopyranose | pH 8.0, 29°C, cosubstrate: 9.1 mM 10-deoxymethynolide | Streptomyces venezuelae | |
5.83 | - |
10-deoxymethynolide | pH 8.0, 29°C, cosubstrate: 10 mM dTDP-3-amino-4,6-dideoxy-alpha-D-glucopyranose | Streptomyces venezuelae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
additional information | - |
the calculated molecular mass of the heterodimer of the DesVII-C-His6/DesVIII-N-Stag complex is 92559 Da | Streptomyces venezuelae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
TDP-3-dimethylamino-4,6-dideoxy-alpha-D-glucopyranose + 10-deoxymethynolide | Streptomyces venezuelae | DesVII is the glycosyltransferase responsible for the attachment of TDP-D-desosamine to macrolactones of varied ring sizes. The enzyme is involved in the biosynthesis of methymycin, neomethymycin, narbomycin, and pikromycin in Streptomyces venezuelae | dTDP + 10-deoxymethymycin | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Streptomyces venezuelae | Q9ZGH7 | - |
- |
Purification (Comment) | Organism |
---|---|
- |
Streptomyces venezuelae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
TDP-3-dimethylamino-4,6-dideoxy-alpha-D-glucopyranose + 10-deoxymethynolide | DesVII is the glycosyltransferase responsible for the attachment of TDP-D-desosamine to macrolactones of varied ring sizes. The enzyme is involved in the biosynthesis of methymycin, neomethymycin, narbomycin, and pikromycin in Streptomyces venezuelae | Streptomyces venezuelae | dTDP + 10-deoxymethymycin | - |
? | |
TDP-3-dimethylamino-4,6-dideoxy-alpha-D-glucopyranose + 10-deoxymethynolide | dTDP-3-amino-4,6-dideoxy-alpha-D-glucopyranose i.e. dTDP-D-desosamine (a 14-membered macrolactone ring). The formation of aglycone is observed when 10-deoxymethymycin is incubated with an excess of TDP in the presence of DesVII-C-His6/DesVIII-N-Stag. The equilibrium of the reaction strongly favors the glycosylation direction because only less than 10% of 10-deoxymethylmycin is converted to 10-deoxymethynolide after a 24 h incubation period | Streptomyces venezuelae | dTDP + 10-deoxymethymycin | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | the DesVII-C-His6/DesVIII-N-Stag complex likely exists as a trimer of heterodimers | Streptomyces venezuelae |
Synonyms | Comment | Organism |
---|---|---|
DesVII | - |
Streptomyces venezuelae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
29 | - |
assay at | Streptomyces venezuelae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0062 | - |
TDP-3-dimethylamino-4,6-dideoxy-alpha-D-glucopyranose | pH 8.0, 29°C, cosubstrate: 9.1 mM 10-deoxymethynolide | Streptomyces venezuelae | |
0.0092 | - |
10-deoxymethynolide | pH 8.0, 29°C, cosubstrate: 10 mM dTDP-3-amino-4,6-dideoxy-alpha-D-glucopyranose | Streptomyces venezuelae | |
1 | - |
10-deoxymethynolide | pH 8.0, 29°C, addition of the DesIII protein, cosubstrate: 1.0 mM dTDP-3-amino-4,6-dideoxy-alpha-D-glucopyranose | Streptomyces venezuelae | |
1.33 | - |
TDP-3-dimethylamino-4,6-dideoxy-alpha-D-glucopyranose | pH 8.0, 29°C, addition of the DesIII protein, cosubstrate: 9.1 mM 10-deoxymethynolide | Streptomyces venezuelae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Streptomyces venezuelae |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
DesIII protein | glycosyltransferase DesVII is unique among glycosyltransferases in that it requires an additional protein component, DesVIII, for activity. desVIII is important for the biosynthesis of glycosylated macrolides but can be replaced by at least one of the homologous genes from other pathways. The addition of recombinant DesVIII protein significantly improves the glycosylation efficiency of DesVII in the in vitro assay. DesVIII assists the folding of DesVII during protein production and remains tightly bound during catalysis. Although the formation of the DesVII/Des-VIII complex is essential for the catalytic activity, DesVIII is unlikely involved in catalysis directly | Streptomyces venezuelae |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00158 | - |
10-deoxymethynolide | pH 8.0, 29°C, cosubstrate: 10 mM dTDP-3-amino-4,6-dideoxy-alpha-D-glucopyranose | Streptomyces venezuelae | |
0.0034 | - |
TDP-3-dimethylamino-4,6-dideoxy-alpha-D-glucopyranose | pH 8.0, 29°C, cosubstrate: 9.1 mM 10-deoxymethynolide | Streptomyces venezuelae | |
0.3125 | - |
TDP-3-dimethylamino-4,6-dideoxy-alpha-D-glucopyranose | pH 8.0, 29°C, addition of the DesIII protein, cosubstrate: 9.1 mM 10-deoxymethynolide | Streptomyces venezuelae | |
16.7 | - |
10-deoxymethynolide | pH 8.0, 29°C, addition of the DesIII protein, cosubstrate: 1.0 mM dTDP-3-amino-4,6-dideoxy-alpha-D-glucopyranose | Streptomyces venezuelae |