Biosynthesis of complex polyketides in a metabolically engineered strain of E. coli.

PFEIFER, B A; ADMIRAL, S J; GRAMAJO, HUGO CÉSAR; CANE, D; KHOSLA, C

SCIENCE

Año: 2001 vol. 291 p. 1790 - 1792

0036-8075

The macrocyclic core of the antibiotic erythromycin, 6-deoxyerythronolide B (6dEB), is a complex natural product synthesized by the soil bacterium Saccharopolyspora erythraea through the action of a multifunctional polyketide synthase (PKS). The engineering potential of modular PKSs is hampered by the limited capabilities for molecular biological manipulation of organisms ( principally actinomycetes) in which complex polyketides have thus far been produced. To address this problem, a derivative of Escherichia coli has been genetically engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. synthase (PKS). The engineering potential of modular PKSs is hampered by the limited capabilities for molecular biological manipulation of organisms ( principally actinomycetes) in which complex polyketides have thus far been produced. To address this problem, a derivative of Escherichia coli has been genetically engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. erythraea through the action of a multifunctional polyketide synthase (PKS). The engineering potential of modular PKSs is hampered by the limited capabilities for molecular biological manipulation of organisms ( principally actinomycetes) in which complex polyketides have thus far been produced. To address this problem, a derivative of Escherichia coli has been genetically engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. synthase (PKS). The engineering potential of modular PKSs is hampered by the limited capabilities for molecular biological manipulation of organisms ( principally actinomycetes) in which complex polyketides have thus far been produced. To address this problem, a derivative of Escherichia coli has been genetically engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. Saccharopolyspora erythraea through the action of a multifunctional polyketide synthase (PKS). The engineering potential of modular PKSs is hampered by the limited capabilities for molecular biological manipulation of organisms ( principally actinomycetes) in which complex polyketides have thus far been produced. To address this problem, a derivative of Escherichia coli has been genetically engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. synthase (PKS). The engineering potential of modular PKSs is hampered by the limited capabilities for molecular biological manipulation of organisms ( principally actinomycetes) in which complex polyketides have thus far been produced. To address this problem, a derivative of Escherichia coli has been genetically engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. through the action of a multifunctional polyketide synthase (PKS). The engineering potential of modular PKSs is hampered by the limited capabilities for molecular biological manipulation of organisms ( principally actinomycetes) in which complex polyketides have thus far been produced. To address this problem, a derivative of Escherichia coli has been genetically engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. Escherichia coli has been genetically engineered. The resulting cellular catalyst converts exogenous propionate into 6dEB with a specific productivity that compares well with a high-producing mutant of S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. over decades for the industrial production of erythromycin. S. erythraea that has been incrementally enhanced over decades for the industrial production of erythromycin. but neither DEBS11TE nor DEBS2 could be purified in detectable quantities. The key parameter that facilitated detection of in vitro activity and subsequent purification of DEBS11TE and DEBS2 was the incubation temperature after IPTG (isopropyl-b-D-thiogalactopyranoside) induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli temperature after IPTG (isopropyl-b-D-thiogalactopyranoside) induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli purified in detectable quantities. The key parameter that facilitated detection of in vitro activity and subsequent purification of DEBS11TE and DEBS2 was the incubation temperature after IPTG (isopropyl-b-D-thiogalactopyranoside) induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli temperature after IPTG (isopropyl-b-D-thiogalactopyranoside) induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli 1TE nor DEBS2 could be purified in detectable quantities. The key parameter that facilitated detection of in vitro activity and subsequent purification of DEBS11TE and DEBS2 was the incubation temperature after IPTG (isopropyl-b-D-thiogalactopyranoside) induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli temperature after IPTG (isopropyl-b-D-thiogalactopyranoside) induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli 1TE and DEBS2 was the incubation temperature after IPTG (isopropyl-b-D-thiogalactopyranoside) induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli b-D-thiogalactopyranoside) induction. Upon lowering the expression temperature from 30° to 22°C, active DEBS11TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coli could be detected in recombinant E. coli 1TE, DEBS2, and DEBS3 proteins could be detected in recombinant E. coliE. coli