INVESTIGADORES
MENZELLA Hugo Gabriel
artículos
Título:
Redesign, synthesis and functional expression of the 6-deoxyerythronolide B polyketide synthase gene cluster
Autor/es:
MENZELLA HG; REISINGER S.; WELCH M.; KEALEY J.T.; KENNEDY J.; REID R.; TRAN C.Q.; SANTI D.
Revista:
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
Editorial:
SPRINGER HEIDELBERG
Referencias:
Año: 2006 vol. 33 p. 22 - 28
ISSN:
1367-5435
Resumen:
A generic design of Type I polyketide synthase genes has been reported
in which modules, and domains within modules, are flanked by sets of
unique restriction sites that are repeated in every module [1]. Using
the universal design, we synthesized the six-module DEBS gene cluster
optimized for codon usage in E. coli, and cloned the three open reading
frames into three compatible expression vectors. With one correctable
exception, the amino acid substitutions required for restriction site
placements were compatible with polyketide production. When expressed in
E. coli the codon-optimized synthetic gene cluster produced
significantly more protein than did the wild-type sequence. Indeed, for
optimal polyketide production, PKS expression had to be down-regulated
by promoter attenuation to achieve balance with expression of the
accessory proteins needed to support polyketide biosynthesis.