Abstract
Toyocamycin exhibits effective biological activities for use against plant pathogenic fungi thanks to its structural similarity to nucleoside. It has been recognized as a promising agricultural antibiotic utilized in controlling the occurrence of plant diseases. In our previous study, a strain that was isolated was identified and designated as Streptomyces diastatochromogenes whose major secondary metabolite was toyocamycin, but the production was largely limited. Protoplast transformation is a useful technique in the improvement of streptomycete. In this study, we optimized some key factors necessary for protoplast formation, regeneration, and transformation of S. diastatochromogenes. When mycelium was cultivated in CP medium with 1 % glycine, harvested at 48 h old, and then treated with 3 mg lysozyme/mL in P buffer for 1 h, the greatest regeneration frequency (42.5 %) of protoplasts was obtained. By using 1 × 109/mL protoplasts with polyethylene glycol 1000 at a concentration of 30 % (w/v), the best performance of protoplast transformation efficiency was 4.8 × 103/μg DNA transformants.
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References
Anne J, Van Mellaert L, Eyssen H (1990) Optimum conditions for efficient transformation of Streptomyces venezuelae protoplasts. Appl Microbiol Biotechnol 32(4):43–435
Apichaisataienchote B, Altenbuchner J, Buchenauer H (2005) Isolation and identification of Streptomyces fradiae SU-1 from Thailand and protoplast transformation with the chitinase B gene from Nocardiopsis prasina OPC-131. Curr Microbiol 51(2):116–121. doi:10.1007/s00284-005-4402-3
Battaglia U, Long JE, Searle MS, Moody CJ (2011) 7-Deazapurine biosynthesis: NMR study of toyocamycin biosynthesis in Streptomyces rimosus using 2-(13)C-7-(15)N-adenine. Org Biomol Chem 9(7):2227–2232. doi:10.1039/c0ob01054e
Bibb MJ, Ward JM, Hopwood DA (1978) Transformation of plasmid DNA into Streptomyces at high frequency. Nature 274(5669):398–400
Furs AR, Orekhov AV (1988) The methods of protoplast formation and transformation of Streptomyces strains. Antibiot Khimioter 33(3):180–185
Hopwood D, Bibb M, Chater K, Kieser T (1985) Genetic manipulation of Streptomyces: a laboratory manual. John Innes Foundation Press, England
Illing GT, Normansell ID, Peberdy JF (1989) Protoplast isolation and regeneration in Streptomyces clavuligerus. J Gen Microbiol 135(8):2289–2297
Iocheva LD, Antonova-Nikolova SK (2000) The effect of protoplast formation on the antibiotic activity and composition of the actinomycin complex in a strain of Streptomyces galbus (F) subsp. achromogenes 695 and in its active variants. Antibiot Khimioter 45(4):10–13
Isono K (1988) Nucleoside antibiotics: structure, biological activity, and biosynthesis. J Antibiot (Tokyo) 41(12):1711–1739
Kieser TBM, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces genetics (2nd ed). John Innes Foundation, England
Lyutzkanova D, Nikolova B, Stoilova-Discheva M, Todorov T (1993) Protoplast formation and regeneration in Streptomyces flavopersicus. Lett Appl Microbiol 16(4):217–219
Malanicheva IA, Koz'mian LI (1990) Effect of protoplast formation and regeneration on the production of nebramycin complex in Streptomyces cremeus subsp. tobramycini. Antibiot Khimioter 35(7):3–5
Matsushima P, Baltz RH (1985) Efficient plasmid transformation of Streptomyces ambofaciens and Streptomyces fradiae protoplasts. J Bacteriol 163(1):180–185
Mellouli L, Karray-Rebai I, Sioud S, Ben Ameur-Mehdi R, Naili B, Bejar S (2004) Efficient transformation procedure of a newly isolated Streptomyces sp. TN58 strain producing antibacterial activities. Curr Microbiol 49(6):400–406
Olano C, Lombo F, Mendez C, Salas JA (2008) Improving production of bioactive secondary metabolites in actinomycetes by metabolic engineering. Metab Eng 10(5):281–292. doi:10.1016/j.ymben.2008.07.001
Shentu XP, Yu B, Bian YL, Hu J, Ma Z, Tong CM, Yu XP (2012) Biocontrol effect and taxonomy of antagonistic Streptomyces strain B28. Acta Phytopathologica Sinica (In Chinese) 42(1):105–109
Tang Z, Xiao C, Zhuang Y, Chu J, Zhang S, Herron PR, Hunter IS, Guo M (2011) Improved oxytetracycline production in Streptomyces rimosus M4018 by metabolic engineering of the G6PDH gene in the pentose phosphate pathway. Enzyme Microb Technol 49(1):17–24. doi:10.1016/j.enzmictec.2011.04.002
Yu B, Shentu X, Yu X (2011) Antifungal activity of toyocamycin on Rhizoctonia solani Kühn. Chinese Journal of Biological Control (In Chinese) 27(3):373–377
Acknowledgments
This work was supported by the Zhejiang key project of innovative team (2009R500273), Zhejiang S&T project (2010C32063), Zhejiang Province Natural Science Foundation (LQ12C14004, LY12C140046), Research on the Public Welfare Technology Application Projects of Zhejiang Province (no. 2013C31028), and National Basic Research Program of China (973 Program) (no. 2011CB111602, 2012CB114100).
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Zheng Ma and Jinxiu Liu contribute equally to this article
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Ma, Z., Liu, J., Lin, X. et al. Formation, regeneration, and transformation of protoplasts of Streptomyces diastatochromogenes 1628. Folia Microbiol 59, 93–97 (2014). https://doi.org/10.1007/s12223-013-0271-5
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DOI: https://doi.org/10.1007/s12223-013-0271-5