INVESTIGADORES
GRAMAJO Hugo Cesar
artículos
Título:
Metabolically engineered Escherichia coli for efficient production of glycosylated natural products
Autor/es:
SALVADOR PEIRÚ, HUGO MENZELLA, EDUARDO RODRÍGUEZ, JOHN CARNEY AND HUGO GRAMAJO
Revista:
Microbial Biotechnology
Editorial:
WILEY BACKWELL
Referencias:
Año: 2008 vol. 1 p. 476 - 486
ISSN:
1751-7915
Resumen:
Significant achievements in polyketide gene expression
have made Escherichia coli one of the most
promising hosts for the heterologous production of
pharmacologically important polyketides. However,
attempts to produce glycosylated polyketides, by the
expression of heterologous sugar pathways, have
been hampered until now by the low levels of glycosylated
compounds produced by the recombinant
hosts. By carrying out metabolic engineering of three
endogenous pathways that lead to the synthesis of
TDP sugars in E. coli, we have greatly improved
the intracellular levels of the common deoxysugar
intermediate TDP-4-keto-6-deoxyglucose resulting in
increased production of the heterologous sugars
TDP-L-mycarose and TDP-D-desosamine, both components
of medically important polyketides. Bioconversion
experiments carried out by feeding
6-deoxyerythronolide B (6-dEB) or 3-a-mycarosylerythronolide
B (MEB) demonstrated that the
genetically modified E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.Escherichia coli one of the most
promising hosts for the heterologous production of
pharmacologically important polyketides. However,
attempts to produce glycosylated polyketides, by the
expression of heterologous sugar pathways, have
been hampered until now by the low levels of glycosylated
compounds produced by the recombinant
hosts. By carrying out metabolic engineering of three
endogenous pathways that lead to the synthesis of
TDP sugars in E. coli, we have greatly improved
the intracellular levels of the common deoxysugar
intermediate TDP-4-keto-6-deoxyglucose resulting in
increased production of the heterologous sugars
TDP-L-mycarose and TDP-D-desosamine, both components
of medically important polyketides. Bioconversion
experiments carried out by feeding
6-deoxyerythronolide B (6-dEB) or 3-a-mycarosylerythronolide
B (MEB) demonstrated that the
genetically modified E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E. coli, we have greatly improved
the intracellular levels of the common deoxysugar
intermediate TDP-4-keto-6-deoxyglucose resulting in
increased production of the heterologous sugars
TDP-L-mycarose and TDP-D-desosamine, both components
of medically important polyketides. Bioconversion
experiments carried out by feeding
6-deoxyerythronolide B (6-dEB) or 3-a-mycarosylerythronolide
B (MEB) demonstrated that the
genetically modified E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.D-desosamine, both components
of medically important polyketides. Bioconversion
experiments carried out by feeding
6-deoxyerythronolide B (6-dEB) or 3-a-mycarosylerythronolide
B (MEB) demonstrated that the
genetically modified E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.a-mycarosylerythronolide
B (MEB) demonstrated that the
genetically modified E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E. coli B strain was able to
produce 60- and 25-fold more erythromycin D (EryD)
than the original strain K207-3, respectively. Moreover,
the additional knockout of the multidrug efflux
pump AcrAB further improved the ability of the engineered
strain to produce these glycosylated compounds.
These results open the possibility of using E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.E.
coli as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.as a generic host for the industrial scale production
of glycosylated polyketides, and to combine
the polyketide and deoxysugar combinatorial approaches
with suitable glycosyltransferases to yield
massive libraries of novel compounds with variations
in both the aglycone and the tailoring sugars.