TAR cloning and integrated overexpression of 6-demethylchlortetracycline biosynthetic gene cluster in Streptomyces aureofaciens

Naxin Wu, He Huang, Taoling Min, and Haifeng Hu

School of Pharmacy, Department of Pharmacology, Fudan University, Shanghai 201203, China

Acta Biochim Biophys Sin 2017, 49: 1129–1134; doi: 10.1093/abbs/gmx110

6-Demethylchlortetracycline (6-DCT), a tetracycline antibiotic produced by Streptomyces aureofaciens, is a crucial precursor employed for the semi-synthesis of tigecycline, minocycline, and amadacyclin (PTK 0796). In this study, the 6-DCT biosynthetic gene cluster (BGC) was cloned from genomic DNA of a high 6-DCT-producing strain, S. aureofaciens DM-1, using the transformation-associated recombination method. An extra copy of the 6-DCT BGC was introduced and integrated into the chromosome of S. aureofaciens DM-1. Duplication of the 6-DCT BGC resulted in a maximum increase of the 6-DCT titer by 34%.

 Direct cloning of the 6-DCT BGC using the TAR method

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