INVESTIGADORES
MENZELLA Hugo Gabriel
artículos
Título:
Production of the potent antibacterial polyketide erythromycin C in Escherichia coli
Autor/es:
PEIRÚ S.; MENZELLA H.G.; RODRÍGUEZ E.; CARNEY J.; GRAMAJO H
Revista:
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Editorial:
AMER SOC MICROBIOLOGY
Referencias:
Año: 2005 vol. 71 p. 2539 - 2547
ISSN:
0099-2240
Resumen:
An Escherichia coli strain capable of producing the potent antibiotic
erythromycin C (Ery C) was developed by expressing 17 new heterologous
genes in a 6-deoxyerythronolide B (6dEB) producer strain. The
megalomicin gene cluster was used as the source for the construction of
two artificial operons that contained the genes encoding the deoxysugar
biosynthetic and tailoring enzymes necessary to convert 6dEB to Ery C.
The reconstructed mycarose operon contained the seven genes coding for
the enzymes that convert glucose-1-phosphate (G-1-P) to TDP-L-mycarose, a
6dEB mycarosyl transferase, and a 6dEB 6-hydroxylase. The activity of
the pathway was confirmed by demonstrating conversion of exogenous 6dEB
to 3-O-alpha-mycarosylerythronolide B (MEB). The reconstructed
desosamine operon contained the six genes necessary to convert
TDP-4-keto-6-deoxyglucose, an intermediate formed in the mycarose
pathway, to TDP-D-desosamine, a desosamine transferase, a 6dEB
12-hydroxylase, and the rRNA methyltransferase ErmE; the last was
required to confer resistance to the host cell upon production of mature
macrolide antibiotics. The activity of this pathway was demonstrated by
conversion of MEB to Ery C. When the mycarose and desosamine operons
were expressed in an E. coli strain engineered to synthesize 6dEB, Ery C
and Ery D were produced. The successful production of Ery C in E. coli
shows the potentiality of this model microorganism to synthesize novel
6-deoxysugars and to produce bioactive glycosylated compounds and also
establishes the basis for the future use of E. coli both in the
production of new glycosylated polyketides and for the generation of
novel bioactive compounds through combinatorial biosynthesis.