Studies on a second and third ring cyclization in anthracycline biosynthesis.
This paper focuses on study of second and third ring cyclization in anthracycline biosynthesis by a heterologous gene expression. Firstly, anthracycline non-producing Streptomyces peucetius mutant, D2 was heterologously complemented to produce daunomycins with plasmids pSgs44 and pSYE66, which contain putative cyclase genes of S. galilaeus and S. nogalater, respectively. A point mutation in the cyclase gene dpsY of D2 has changed glycine to serine resulting inactivation of the enzyme. Secondly, the putative cyclase gene snoaM from S. nogalater, was expressed in a gene cassette in S. lividans TK24 and S. coelicolor CH999 to study the influence of the cyclase gene on auramycinone production and the impact of endogenous genes on production profiles. The results obtained confirms that a cyclase closing the second and third ring of a polyketide is essential in anthracycline biosynthesis.
Hautala
Anne
A
Galilaeus Oy, P.O. Box 113, FIN-20781, Kaarina, Finland. anne.hautala@galilaeus.fi
Torkkell
Sirke
S
Räty
Kaj
K
Kunnari
Tero
T
Kantola
Jaana
J
Mantsälä
Pekka
P
Hakala
Juha
J
Ylihonko
Kristiina
K
D018943:Anthracyclines / Q000378:metabolism*; D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D003500:Cyclization; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D016254:Mutagenesis, Insertional; D019906:Nuclear Magnetic Resonance, Biomolecular; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*
D018943:Anthracyclines; D000903:Antibiotics, Antineoplastic; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C112173:DpsY protein, Streptomyces peucetius; D011994:Recombinant Proteins