Abstract
The epothilones are highly promising prospective anticancer agents that are produced by the myxobacterium Sorangium cellulosum. We mutated the epothilone producing S. cellulosum strain So0157-2 to improve the production of epothilones. For evaluation in high-throughput of a large number of mutants, we developed a simple microtiter method for primary screening. Using the classical UV-mutation method plus selection pressures, the production capacity was increased about 0.5∼2.5 times the starting strain. The mutants with higher production and different phenotypes were further subjected to recursive protoplast fusions and the fusants products were screened under multi-selection pressure. Furthermore, the production was greatly increased by the genome shuffling. For epothilone B, the production of one fusant was increased about 130 times compared to the starting strain, increasing from 0.8 mg l−1 to 104 mg l−1.
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Acknowledgments
This work was financially supported by grants 30370792, 30400009, 30671192 of Chinese National Natural Science Foundation. The authors thank Dr. Roberta Greenwood for her help in editing this manuscript.
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Guo-li Gong and Xin Sun contributed equally to this work.
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Gong, Gl., Sun, X., Liu, Xl. et al. Mutation and a high-throughput screening method for improving the production of Epothilones of Sorangium . J Ind Microbiol Biotechnol 34, 615–623 (2007). https://doi.org/10.1007/s10295-007-0236-2
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DOI: https://doi.org/10.1007/s10295-007-0236-2