Knockout of N-hydroxybenzoate hydroxylase functional gene orf54 in Streptomyces sp. 13580 gene cluster and fermentation detection of its mutant strain

Knockout of the N-hydroxybenzoate hydroxylase functional gene orf54 in the Streptomyces sp. 13580 gene cluster and its fermentation HPLC detection of its mutant strain. We identified the biosynthetic gene cluster from the strain Streptomyces sp. 13580 by genome sequencing, and finding genes for hydroxylase function through bioinformatics analysis. The cosmid Δorf54 deleting the N-hydroxybenzoate hydroxylase functional gene orf54 was constructed in Escherichia coli by PCR-targeting. Transfer of the mutant cosmid into Streptomyces sp. 13580. The orf54 gene sequence was knocked out by homologous recombination, and the double-crossover insertion-inactivated mutant 13580 Δorf54 was screened. The wild type strain Streptomyces sp. 13580 and its mutant 13580 Δorf54 were subjected to shake flask fermentation and then analyzed by high performance liquid chromatography (HPLC).