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2025年5月7日 星期三
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提高苏云金素合成纯度的发酵工艺的优化
孙明 1,刘要2
1. 华中农业大学农业微生物学国家重点实验室
2. 华中农业大学生命科学技术学院,农业微生物学国家重点实验室,武汉 430070
The optimization of fermentation process to increase purity of thuringiensin in the extract from supernatant liquid
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摘要 摘要:苏云金芽胞杆菌中华亚种CT-43菌株(Bacillus thuringiensis subsp.chinensis)是高产苏云金素(简称Thu)的野生菌株。但在产生Thu时总伴随有其他代谢物(简称ZZ)的合成,而且ZZ很难与Thu分离开。本实验通过比较不同的培养基,得到G培养基能抑制菌株CT-43合成ZZ。然后利用分批补料发酵的方法,通过正交试验设计,进一步抑制ZZ的合成,并提高Thu在有机萃取物中的含量。得到优化后的发酵工艺:菌株CT-43在G培养基中培养至第20 h,同时补柠檬酸钠和葡萄糖至终浓度分别为2.0g/L和16g/L,再继续发酵至第52h。通过此发酵工艺制备的Thu峰面积占初提物总峰面积74.84,比用LB培养基发酵所得提取物的29.85%提高了一倍多。为得到高纯度的Thu和更深入的研究创造了条件。最后通过液质联用技术证实,发酵工艺优化后,菌株CT-43发酵上清液提取物中主要物质是Thu,几乎检测不到ZZ。
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孙明
刘要
关键词 苏云金芽胞杆菌苏云金素补料发酵正交试验设计纯化液质联用    
Abstract:Abstract: B. thuringiensis subsp.chinensis strain CT-43 highly produces thuringiensin (Thu). Meanwhile, other metabolizing product (ZZ) was synthesized, but was difficultly removed from Thu. In this study, various media were screened and the G medium was found to inhibite the ZZ synthesis. Based on the G medium, feed-batch fermentation and orthogonal arrays was combined to further inhibit ZZ synthesis and increase the yield of Thu.. The optimal fermentation process was that, After strain CT-43 was cultured in G medium for 20h, patching with 2.0 g/L sodium citrate and 16 g/L glucose and culturing for additional 32h, the peak area ratio of Thu could get 74.84 in the primary extract, which was 2.5 times for 29.85% of LB fermentation. These onditions could create a favorable fermention process to produce more pure Thu and it will enefit for further research. Lastly, HPLC-MS was used to verify the produced Thu. Thu became he major element during the feed-batch fermentation. No ZZ was detected and Thu was not modified after above fermentation process.
收稿日期: 2009-04-22     
通讯作者: 孙明    
引用本文:   
孙明 1,刘要2. 提高苏云金素合成纯度的发酵工艺的优化[J]. , 2010, 18(2): 406-412.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2010/V18/I2/406
 
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