Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 83, Issue 3, Pages 507-512Publisher
SPRINGER
DOI: 10.1007/s00253-009-1918-7
Keywords
Protoplast fusion; Streptomyces hygroscopicus; Rapamycin; Genome shuffling
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Funding
- National High Technology Research and Development Program of China [2006AA02Z239]
- National Basic Research Program of China [2007CB707805]
- Ministry of Science and Technology of China
- National Science Foundation of China [20736008, 20676115]
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Rapamycin is a 31-member ring macrolide produced by Streptomyces hygroscopicus and has many applications in clinical medicine. In the present work, several protoplasts-related techniques including protoplasts mutation, intraspecies and interspecies protoplasts fusion were tried to improve the rapamycin productivity in S. hygroscopicus. Although mutation and fusion of different protoplasts of S. hygroscopicus did not improve the productivity of rapamycin significantly, the interspecies fusion of protoplasts of S. hygroscopicus D7-804 and Streptomyces erythreus ZJU325 could have brought about one high-yield (345 mg/L) rapamycin producer with 23.6% higher than that of the parental strain. Then, with seven mutants of S. hygroscopicus with different features and rapamycin productivities as the parental strains, only one-round genome shuffling has generated a high-yield rapamycin-producing strain with an outstanding yield of 445 mg/L. The systematic research of protoplast-related techniques has established an applicable way to generate high-yield strains from original microorganisms which can only produce low amount of expected natural products, without information of target gene clusters and gene sequences.
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