title: Streptomycin resistance-aided genome shuffling to improve doramectin productivity of Streptomyces avermitilis NEAU1069.
abstract: Genome shuffling is an efficient approach for the rapid engineering of microbial strains with desirable industrial phenotypes. In this study, a strategy of incorporating streptomycin resistance screening into genome shuffling (GS-SR) was applied for rapid improvement of doramectin production by Streptomyces avermitilis NEAU1069. The starting mutant population was generated through treatment of the spores with N-methyl-N'-nitro-N-nitrosoguanidine and ultraviolet (UV) irradiation, respectively, and five mutants with higher productivity of doramectin were selected as starting strains for GS-SR. Finally, a genetically stable strain F4-137 was obtained and characterized to be able to yield 992 ± 4.4 mg/l doramectin in a shake flask, which was 7.3-fold and 11.2-fold higher than that of the starting strain UV-45 and initial strain NEAU1069, respectively. The doramectin yield by F4-137 in a 50-l fermentor reached 930.3 ± 3.8 mg/l. Furthermore, the factors associated with the improved doramectin yield were investigated and the results suggested that mutations in ribosomal protein S12 and the enhanced production of cyclohexanecarboxylic coenzyme A may contribute to the improved performance of the shuffled strains. The random amplified polymorphic DNA analysis showed a genetic diversity among the shuffled strains, which confirmed the occurrence of genome shuffling. In conclusion, our results demonstrated that GS-SR is a powerful method for enhancing the production of secondary metabolites in Streptomyces.
authors: College of Life Science, Northeast Agricultural University, Harbin, 150030, People's Republic of China.
authors: Ji
authors: J
authors: Zhang
authors: Xiangjing
authors: X
authors: Wang
authors: Jinna
authors: J
authors: Diao
authors: Hairong
authors: H
authors: He
authors: Yuejing
authors: Y
authors: Zhang
authors: Wensheng
authors: W
authors: Xiang
mesh_terms: D000871:Anthelmintics / Q000737:chemistry / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D019149:Bioreactors; D032621:DNA Shuffling / Q000379:methods*; D024881:Drug Resistance, Bacterial; D005285:Fermentation; D014644:Genetic Variation; D016680:Genome, Bacterial; D007559:Ivermectin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D009154:Mutation; D013302:Streptomyces / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013307:Streptomycin / Q000494:pharmacology*
chemical_list: D000871:Anthelmintics; D000900:Anti-Bacterial Agents; D007559:Ivermectin; C084101:doramectin; D013307:Streptomycin
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