Abstract
Among eight components of avermectin, B1 fractions have the most effective antiparasitic activities and the lowest level of toxic side-effects and are used widely in veterinary and agricultural fields. Intraspecific protoplast fusion between two strains of Streptomyces avermitilis, one an avermectin high producer (strain 76-05) and the other a genetically engineered strain containing the mutations aveD − and olmA − (strain 73-12) was performed for enhancement and selective production of avermectin B in the absence of oligomycin. Two recombinant strains (F23 and F29) were isolated and characterized with regards to the parental merits. F23 and F29 produced only the four avermectin B components with high yield and produced no oligomycin. The avermectin production of F23 and F29 was about 84.20% and 103.45% of the parental strain 76-05, respectively, and increased about 2.66-fold and 3.50-fold, respectively, compared to that of parental strain 73-12. F23 and F29 were genetically stable prototrophic recombinants and F29 was quite tolerant of fermentation conditions compared to avermectin high producer parental strain 76-05. The ability to produce avermectin B with high yield without the production of other avermectin components and oligomycin will make F23 and F29 useful strains for avermectin production. Strain F29’s tolerance of fermentation conditions will also make it suitable for industrial applications.
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References
Burg R W, Miller B M, Baker E E, et al. Avermectins, new family of potent anthelmintic agents: Producing organisms and fermentation. Antimicrob Agents Chemother, 1979, 15: 361–367
Egerton J R, Ostling D A, Blair L S, et al. Avermectins, new family of potent anthelmintic agents: Efficacy of the B1a component. Antimicrob Agents Chemother, 1979, 15: 372–378
Pinna L A, Lorini M, Moret V, et al. Effect of oligomycin and succinate on mitochondrial metabolism of adenine nucleotides. Biochim Biophys Acta, 1967, 143: 18–25
Ikeda H, Ōmura S. Control of avermectin biosynthesis in Streptomyces avermitilis for the selective production of a useful component. J Antibiot, 1995, 48(7): 549–562
Ikeda H, Nonomiya T, Usami M, et al. Organization of the biosynthetic gene cluster for the polyketide anthelmintic macrolide avermectin in Streptomyces avermitilis. Proc Natl Acad Sci USA, 1999, 96(17): 9509–9514
Ikeda H, Ishikawa J, Hanamoto A, et al. Complete genome sequence and comparative analysis of the industrial microorganism Streptomyces avermitilis. Nat Biotechnol, 2003, 21: 526–531
Ōmura S, Ikeda H, Ishikawa J, et al. Genome sequence of an industrial microorganism Streptomyces avermitilis: Deducing the ability of producing secondary metabolites. Proc Natl Acad Sci USA, 2001, 98(21): 12215–12220
Chen Z, Wen Y, Song Y, et al. Effect of gene deletion of aveD on avermectins production in Streptomyces avermitilis. Acta Microbiol Sinica (in Chinese), 2002, 42(5): 534–538
Zhang X L, Chen Z, Zhao J L, et al. Deletion analysis of oligomycin PKS genes (olmA) in Streptomyces avermitilis. Chin Sci Bull, 2004, 49: 350–354
Zhou X F, Zhou Q. Interspecific protoplast fusion of Streptomyces hygroscopicus var. yingchengenisis with Streptomyces qingfengmyceticus and biological characterization of their recombinants. Chin J Biotechnol, 1989, 5:161–166
Baltz R H. Genetic recombination by protoplast fusion in Streptomyces. J Ind Microbiol Biot, 1999, 22: 460–471
Hopwood D A, Wright H M. Bacterial protoplast fusion: recombination in fused protoplasts of Streptomyces coelicolor. J Gen Microbiol, 1978, 162: 307–317
Zhang Y X, Perry K, Vinci V A, et al. Genome shuffling leads to rapid phenotypic improvement in bacteria. Nature, 2002, 415: 644–646
Patnaik R, Louie S, Gavrilovic V, et al. Genome shuffling of Lactobacillus for improved acid tolerance. Nat Biotechnol, 2002, 20: 707–712
Ikeda H, Kotaki H, Tanaka H, et al. Involvement of glucose catabolism in avermectin production by Streptomyces avermitilis. Antimicrob Agents Chemother, 1988, 32: 282–284
MacNeil D J, Klapko L M. Transformation of Streptomyces avermitilis by plasmid DNA. J Ind Microbiol, 1987, 2: 209–218
Song Y, Cao G M, Chen Z, et al. Selection of high avermectins producing strain and identification of avermectin B1. Chin J Biotechnol (in Chinese), 2000, 16(1): 31–35
Kieser T, Bibb M J, Buttner M J, et al. Practical Streptomyces Genetics. Norwich: The John Innes Foundation, 2000
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Supported by National Basic Research Project (Grant No. 2003CB114205) and Key Technologies R&D Programme (Grant No. 2004BA713B02-03)
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Chen, Z., Wen, J., Song, Y. et al. Enhancement and selective production of avermectin B by recombinants of Streptomyces avermitilis via intraspecific protoplast fusion. CHINESE SCI BULL 52, 616–622 (2007). https://doi.org/10.1007/s11434-007-0119-y
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DOI: https://doi.org/10.1007/s11434-007-0119-y