DNA shuffling of ptr resistance gene leads to improved pristinamycin production in Streptomyces pristinaespiralis

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• 作者：Q. -C. Jin ; N. Shen ; H. Yin ; Y. Yang ; Z. -H. Jin
• 关键词：Streptomyces pristinaespiralis ; pristinamycin biosynthesis ; DNA shuffling ; ptr gene
• 刊名：Molecular Biology
• 出版年：2015
• 出版时间：March 2015
• 年：2015
• 卷：49
• 期：2
• 页码：253-259
• 全文大小：1,846 KB
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• 作者单位：Q. -C. Jin (1)
  N. Shen (1)
  H. Yin (1) (2)
  Y. Yang (1)
  Z. -H. Jin (1) (2)
  
  1. School of Biological and Chemical Engineering, Ningbo Institute of Technology, Zhejiang University, Ningbo, 315100, China
  2. Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou, 310027, China
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Life Sciences
  Biochemistry
  Human Genetics
  Russian Library of Science
  
• 出版者：MAIK Nauka/Interperiodica distributed exclusively by Springer Science+Business Media LLC.
• ISSN：1608-3245

文摘

In order to enhance pristinamycin production, six homologous ptr genes from high pristinamycin-producing strains of Streptomyces pristinaespiralis were selected for DNA shuffling, and the reason for the altered activities of the shuffled ptr gene was speculated by sequence alignment. The highest pristinamycin yield of 0.12 g/L was achieved with a sixfold increase in strain sps16 obtained by DNA shuffling when compared to ancestral strain ATCC 25486. Sequence analysis of the ptr gene variant from the sps16 strain indicated that five mutations (H16P, N63D, T75P, Q107R, and P435A) were introduced into the gene, two of them (N63D and T75P) located in the second of the 14 transmembrane segments (TMS). Prediction of the secondary structure of the gene product indicated that mutations at the N-terminus resulted in the shortening of the corresponding α-helix, while the mutation at the C-terminus lengthened the helix. In conclusion, combination of DNA shuffling with genome shuffling is an effective breeding strategy for increasing the antibiotic yield by directed evolution of target genes.