Vol 49 No. 12: 1129-1134 [PDF] [Full Text]
TAR cloning and integrated overexpression of 6-demethylchlortetracycline biosynthetic gene cluster in Streptomyces aureofaciens
Naxin Wu1,2, He Huang3, Taoling Min2, and Haifeng Hu2,*

1School of Pharmacy, Department of Pharmacology, Fudan University, Shanghai 201203, China,
2Shanghai Institute of Pharmaceutical Industry, Department of Biopharmceutical, Shanghai 201203, China, and
3Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China

Abstract  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%.


Keywords   6-demethylchlortetracycline, Streptomyces aureofaciens, biosynthetic gene cluster, transformation-associated recombination


Received   2017-5-24  


Funding  This work was supported by the grants from the Natural Science Foundation of Shanghai (No. 15ZR1440300) and the National Science and Technology Major Project (No. 2014ZX09201-001-05).


* Correspondence address  Tel/Fax: +86-21-62892873; E-mail: haifenghu88@163.com

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