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Microbiology 145 (1999), 855-868; DOI  10.1099/13500872-145-4-855
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The tylosin biosynthetic cluster from Streptomyces fradiae: genetic organization of the left region

Roberto Fouces, Encarnación Mellado, Bruno Díez and José Luis Barredo

Laboratorio de Ingeniería Genética, Antibióticos SA, Avenida de Antibióticos 59–61, 24080 León, Spain

Author for correspondence: José Luis Barredo. Tel: +34 987 895826. Fax: +34 987 895810. e-mail: jbarredo@antibioticos.it

ABSTRACT

Summary: The genetic organization of the left edge (tylEDHFJ region) of the tylosin biosynthetic gene cluster from Streptomyces fradiae has been determined. Sequence analysis of a 12·9 kb region has revealed the presence of 11 ORFs, 10 of them belonging to the biosynthetic cluster. The putative functions of the proteins encoded by these genes are as follows: peptidase (ORF1, ddcA), tylosin resistance determinant (ORF2, tlrB), glycosyltransferase (ORF3, tylN), methyltransferase (ORF4, tylE), ketoreductase (ORF5, tylD), ferredoxin (ORF6, tylH2), cytochrome P450 (ORF7, tylH1), methyltransferase (ORF8, tylF), epimerase (ORF9, tylJ), acyl-CoA oxidase (ORF10, tylP) and receptor of regulatory factors (ORF11, tylQ). The functional identification of the genes in the proposed tylosin biosynthetic pathway has been deduced by database searches and previous genetic complementation studies performed with tylosin idiotrophic mutants blocked at various stages in tylosin biosynthesis. The tlrB gene has been shown to be useful as a tylosin resistance marker in Streptomyces lividans, Streptomyces parvulus and Streptomyces coelicolor and the effect of tylF on macrocin depletion has been confirmed. A pathway for the biosynthesis of 6-deoxy-D-allose, the unmethylated mycinose precursor, involving the genes tylD, tylJ and tylN is proposed.

Keywords: glycosyltransferase, ketoreductase, cytochrome P450, methyltransferase, mycinose




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