A sequential statistical approach was applied to optimizing the fermentation medium of epothilones（Epos） production by means of a mutant which was obtained by treating polyangium cellulosum ATCC 15384 with nitrite a...A sequential statistical approach was applied to optimizing the fermentation medium of epothilones（Epos） production by means of a mutant which was obtained by treating polyangium cellulosum ATCC 15384 with nitrite and ultraviolet. The effects of different carbon sources and nitrogen sources on the fermentation medium were tested, and the suitable ones were selected. Then a uniform design was employed to design the experiments. A linear model was developed for identifying the significant components in fermentation medium, while a third degree polynomial model was used for studying the relationship between the concentration of the components in fermentation medium and the yield of Epos（YEPs）. A pattern search method was used for searching the optimum fermentation medium in the test space, which was as follows（g/L）： potassium nitrate 8.00, soybean peptone 17.60, potassium hydrogen phos- phate 1.00, beef extraction 6.46, yeast extraction 1.00, calcium chloride 0.25, sodium chloride 1.00 and ferric chloride 0.02. The optimum fermentation medium was expected to result in a yield of Epos（YEPs） of 2.48 mg/L. The validation experiments with the optimum medium were performed in triplicate and the average yield of Epos was 2.45 mg/L which was 7.78 times higher than that of Epos prepared without optimization.展开更多
Endusamycin,a naturally occurring polyether ionophore antibiotic,exhibits extensive antitumor activities.Despite its promising potential,the titer of endusamycin is significantly lower compared to widely used polyethe...Endusamycin,a naturally occurring polyether ionophore antibiotic,exhibits extensive antitumor activities.Despite its promising potential,the titer of endusamycin is significantly lower compared to widely used polyether compounds,and no reports have been published regarding its overproduction.In this study,various metabolic engineering strategies were performed to enhance endusamycin production.Notably,the deletion of competing biosynthetic gene clusters(BGCs)responsible for the biosynthesis of spore pigment and meilingmycin-like compounds based on transcriptome analysis,as well as the doubling of the endusamycin BGC,proved to be effective.These interventions resulted in a 20%and 69%increase in the titer of endusamycin,respectively.Furthermore,systematic optimization of fermentation medium components,including carbon source,nitrogen source,phosphorus and potassium,contributed to a further 69%increase in the titer of endusamycin.Ultimately,the high-yielding strain YC1109 was developed through the integration of these strategies.The titer of endusamycin reached 5469 mg/L in shake-flask fermentation and 5011 mg/L in fed-batch fermentation,representing a 246%increase compared to the original strain.This research significantly facilitates the drug development and industrialization of endusamycin.It establishes a superior chassis strain for exploring endusamycin derivatives and provides valuable insights into improving the production of polyether compounds.展开更多
基金Supported by the Science Technology Development Project of Jilin Province,China(No.20020503-2)
文摘A sequential statistical approach was applied to optimizing the fermentation medium of epothilones（Epos） production by means of a mutant which was obtained by treating polyangium cellulosum ATCC 15384 with nitrite and ultraviolet. The effects of different carbon sources and nitrogen sources on the fermentation medium were tested, and the suitable ones were selected. Then a uniform design was employed to design the experiments. A linear model was developed for identifying the significant components in fermentation medium, while a third degree polynomial model was used for studying the relationship between the concentration of the components in fermentation medium and the yield of Epos（YEPs）. A pattern search method was used for searching the optimum fermentation medium in the test space, which was as follows（g/L）： potassium nitrate 8.00, soybean peptone 17.60, potassium hydrogen phos- phate 1.00, beef extraction 6.46, yeast extraction 1.00, calcium chloride 0.25, sodium chloride 1.00 and ferric chloride 0.02. The optimum fermentation medium was expected to result in a yield of Epos（YEPs） of 2.48 mg/L. The validation experiments with the optimum medium were performed in triplicate and the average yield of Epos was 2.45 mg/L which was 7.78 times higher than that of Epos prepared without optimization.
基金supported by Joint Fund of the National Natural Science Foundation of China[grant number U23A20527]Wuhan Hesheng Technology Co.,Ltd.and Key R&D Program of Hubei Jiangxia Laboratory[grant number E4JXBS0001].
文摘Endusamycin,a naturally occurring polyether ionophore antibiotic,exhibits extensive antitumor activities.Despite its promising potential,the titer of endusamycin is significantly lower compared to widely used polyether compounds,and no reports have been published regarding its overproduction.In this study,various metabolic engineering strategies were performed to enhance endusamycin production.Notably,the deletion of competing biosynthetic gene clusters(BGCs)responsible for the biosynthesis of spore pigment and meilingmycin-like compounds based on transcriptome analysis,as well as the doubling of the endusamycin BGC,proved to be effective.These interventions resulted in a 20%and 69%increase in the titer of endusamycin,respectively.Furthermore,systematic optimization of fermentation medium components,including carbon source,nitrogen source,phosphorus and potassium,contributed to a further 69%increase in the titer of endusamycin.Ultimately,the high-yielding strain YC1109 was developed through the integration of these strategies.The titer of endusamycin reached 5469 mg/L in shake-flask fermentation and 5011 mg/L in fed-batch fermentation,representing a 246%increase compared to the original strain.This research significantly facilitates the drug development and industrialization of endusamycin.It establishes a superior chassis strain for exploring endusamycin derivatives and provides valuable insights into improving the production of polyether compounds.
