Scholarlink

<p>Carrimycin, a new drug approved in recent years, is produced by recombinant <i>Streptomyces spiramyceticus</i> harboring a heterogeneous 4″-<i>O</i>-isovaleryltransferase gene (<i>ist</i>). The regulatory gene <i>bsm42</i> plays a crucial role in activation of carrimycin biosynthesis, its high expression can improve the carrimycin production. The <i>xylE</i>-<i>neo</i> reporter cassette under control of the the two different lengths of <i>bsm42</i> promoters, F1R and F2R, can be used to monitor the expression of <i>bsm42</i> to screen carrimycin high-yield mutants treated with the composite mutagenesis of plasma and ultraviolet radiation. We found that 47% of the mutants selected based on the higher expression of reporter genes driven by <i>bsm42</i> promoters showed improved carrimycin production, and the shorter promoter F2R was more effective than promoter F1R. The high-yield mutant F2R-15 was about 13 times higher than that of 54-IA strain, and its titer reached 1010 ± 30&#xa0;µg/mL. A comparative analysis of the transcriptome profiles between the F2R-15 mutant and 54-IA strains revealed 158 significantly differentially expressed genes. The upregulated genes were associated with macrolide precursor biosynthesis, macrolide inactivation, antibiotic transporters, and oxidative phosphorylation, while the most downregulated genes were related to primary biosynthetic metabolites and other secondary metabolites. These results demonstrate that the reporter-guided selection method combined with traditional mutagenesis is an effective strategy for obtaining stable carrimycin high-yield mutants.</p> Graphical Abstract <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Enhancement of carrimycin production in Streptomyces spiramyceticus 54-IA by the reporter-guided selection method with traditional mutagenesis

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年8月6日

查看全文
  • Kemeng Li,
  • Jianlu Dai,
  • Yue Chang,
  • Tianyi Hao,
  • Weiqing He

摘要

Carrimycin, a new drug approved in recent years, is produced by recombinant Streptomyces spiramyceticus harboring a heterogeneous 4″-O-isovaleryltransferase gene (ist). The regulatory gene bsm42 plays a crucial role in activation of carrimycin biosynthesis, its high expression can improve the carrimycin production. The xylE-neo reporter cassette under control of the the two different lengths of bsm42 promoters, F1R and F2R, can be used to monitor the expression of bsm42 to screen carrimycin high-yield mutants treated with the composite mutagenesis of plasma and ultraviolet radiation. We found that 47% of the mutants selected based on the higher expression of reporter genes driven by bsm42 promoters showed improved carrimycin production, and the shorter promoter F2R was more effective than promoter F1R. The high-yield mutant F2R-15 was about 13 times higher than that of 54-IA strain, and its titer reached 1010 ± 30 µg/mL. A comparative analysis of the transcriptome profiles between the F2R-15 mutant and 54-IA strains revealed 158 significantly differentially expressed genes. The upregulated genes were associated with macrolide precursor biosynthesis, macrolide inactivation, antibiotic transporters, and oxidative phosphorylation, while the most downregulated genes were related to primary biosynthetic metabolites and other secondary metabolites. These results demonstrate that the reporter-guided selection method combined with traditional mutagenesis is an effective strategy for obtaining stable carrimycin high-yield mutants.

Graphical Abstract