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Journal of Bioprocessing & Biotechniques

ISSN: 2155-9821

Open Access

Introduction of Extra Copy of Oxytetracycline Resistance Gene otrB Enhances the Biosynthesis of Oxytetracycline in Streptomyces rimosus

Abstract

Xiaohe Chu, Zijing Zhen, Zhenyu Tang, Yingping Zhuang, Ju Chu, Siliang Zhang and Meijin Guo

The aromatic polyketide antibiotic oxytetracycline (OTC) is produced by Streptomyces rimosus as an important secondary metabolite. Enhancement of self-resistance is one effective way to improve antibiotic production in Streptomyces spp. In the present study, we aimed to improve the production of OTC by introducing extra copies of the OTC resistance genes, otrA and otrB, into the chromosome of the industrial strain of S. rimosus (SRI). First, otrA and otrB were cloned and ligated with pSET152 to generate the recombinant plasmids pSET152-otrA/otrB, the demethylated pSET152-otrA/otrB by Escherichia coli ET12567(pUZ8002) were then introduced into SRI to yield otrA/ otrB knock-in mutants: SRI-A (otrA) and SRI-B (otrB). Ten selected mutants and the parent SRI strain were cultured in shake flasks. Production of OTC was increased by 67% in one SRI-B mutant compared with the parent strain, suggesting that the enhancement of resistance gene otrB in the antibiotic producer is an effective way to improve OTC biosynthesis. However, introduction of extra copy of otrB could retard growth of mutant cells.

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