Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 2.4.1.277 extracted from

  • Borisova, S.A.; Liu, H.W.
    Characterization of glycosyltransferase DesVII and its auxiliary partner protein DesVIII in the methymycin/picromycin biosynthetic pathway (2010), Biochemistry, 49, 8071-8084.
    View publication on PubMedView publication on EuropePMC

Application

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(Commentary)

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 [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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Streptomyces venezuelae Q9ZGH7
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Streptomyces venezuelae

Substrates and Products (Substrate)

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

Subunits Comment Organism
More the DesVII-C-His6/DesVIII-N-Stag complex likely exists as a trimer of heterodimers Streptomyces venezuelae

Synonyms

Synonyms Comment Organism
DesVII
-
Streptomyces venezuelae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
29
-
assay at Streptomyces venezuelae

Turnover Number [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
assay at Streptomyces venezuelae

Cofactor

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 [mM/s]

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