Engineered Biosynthesis of a Complete Macrolactone in a Heterologous Host

• Published in: Science (American Association for the Advancement of Science) Vol. 265; no. 5171; pp. 509 - 512
• Main Authors: Kao, Camilla M., Katz, Leonard, Khosla, Chaitan
• Format: Journal Article
• Language: English
• Published: WASHINGTON American Society for the Advancement of Science 22.07.1994; Amer Assoc Advancement Science; American Association for the Advancement of Science
• Subjects: Acetates; Active sites; Acyl Coenzyme A - metabolism; Base Sequence; Binding Sites; Biochemistry; Bioconversions. Hemisynthesis; Biological and medical sciences; Biosynthesis; Biotechnology; Chemical engineering; Cloning, Molecular; Drug Design; Enzymes; Erythromycin - analogs & derivatives; Erythromycin - biosynthesis; Erythromycin - isolation & purification; Escherichia coli; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Genetic Engineering; Genetic Vectors; Mass spectroscopy; Methods. Procedures. Technologies; Molecular Sequence Data; Multidisciplinary Sciences; Multienzyme Complexes - chemistry; Multienzyme Complexes - genetics; Multienzyme Complexes - metabolism; Multigene Family; Mutation; Oleandomycin - analogs & derivatives; Oleandomycin - biosynthesis; Oleandomycin - isolation & purification; Plasmids; Polyketides; Recombinant Proteins - metabolism; Saccharopolyspora erythraea; Science & Technology; Science & Technology - Other Topics; Streptomyces - enzymology; Streptomyces - genetics; Streptomyces coelicolor; Structure-Activity Relationship; PHYSICOCHEMICAL PROPERTIES; PROPIONIBACTERIUM-SHERMANII; POLYKETIDE SYNTHASE; ESCHERICHIA-COLI; COENZYME-A MUTASE; SACCHAROPOLYSPORA-ERYTHRAEA; MACROLIDE ANTIBIOTICS; METHYLMALONYL-COA EPIMERASE; STREPTOMYCES-ERYTHREUS; NUCLEOTIDE-SEQUENCE; Actinomycetales; Heterologous system; Streptomycetaceae; Genetic transformation; Streptomyces coelicolor; Aglycone; Bacteria; Actinomycetes; Biosynthesis; Recombinant DNA; Erythromycin; Macrolide
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.8036492

Macrocyclic polyketides have been subjects of great interest in synthetic and biosynthetic chemistry because of their structural complexity and medicinal activities. With expression of the entire 6-deoxyerythronolide B synthase (DEBS) (10,283 amino acids) in a heterologous host, substantial quantities of 6-deoxyerythronolide B (6dEB), the aglycone of the macrolide antibiotic erythromycin, and 8,8a-deoxyoleandolide, a 14-membered lactone ring identical to 6dEB except for a methyl group side chain in place of an ethyl unit, were synthesized in Streptomyces coelicolor. The biosynthetic strategy utilizes a genetic approach that facilitates rapid structural manipulation of DEBS or other modular polyketide synthases (PKSs), including those found in actinomycetes with poorly developed genetic methods. From a technological viewpoint, this approach should allow the rational design of biosynthetic products and may eventually lead to the generation of diverse polyketide libraries by means of combinatorial cloning of naturally occurring and mutant PKS modules.