摘要
为了提高两性霉素B(amphotericin B,AmB)的产量,降低生产成本,满足市场需求,采用紫外诱变和常压室温等离子体(atmospheric and room temperature plasma,ARTP)诱变的复合诱变方法进行菌株选育。首先,对结节链霉菌出发菌株A318-10进行复合诱变;然后,筛选高产菌株,并利用软件Design-Expert 12对显著效应因子进行Box-Behnken Design试验设计,得到最优发酵配方;最后,在1 L摇瓶及50 L发酵罐中进行发酵优化配方验证。结果表明,在紫外诱变60 s并ARTP诱变35 s后,筛选得到了高产菌株20-2-15,其最优发酵配方为葡萄糖94.6 g/L、中温豆粉27.7 g/L、淀粉24.1 g/L;在最优配方下,结节链霉菌1 L摇瓶和50 L发酵罐的发酵最高水平分别为18968μg/mL和21257μg/mL,比原始配方分别提高了32.6%和29.4%。研究结果应用于规模化生产后,有利于降低成本、提高企业竞争力。
In order to increase the yield of amphotericin B(AmB),reduce production costs and meet market demand,the combination of ultraviolet mutagenesis and atmospheric and room temperature plasma(ARTP)mutagenesis was used to select the strains.First,complex mutagenesis was performed on the starting strain A318-10 of Streptomyces nodosus.Then,the high-yield strains were screened and the optimal fermentation formula was obtained by Box-Behnken Design test on the significant effect factors using Design-Expert 12.Finally,fermentation optimization formula was verified in 1 L shaker and 50 L fermenter.The results show that the high-yielding strain 20-2-15 is obtained after ultraviolet mutagenesis for 60 s and ARTP mutagenesis for 35 s.The optimal fermentation formula is 94 . 6 g/L glucose,27 . 7 g/L medium temperature soybean powder and 24 . 1 g/L starch.Under the optimal formula,the maximum fermentation levels of 1 L shaker and 50 L fermenter are 18968μg/mL and 21257μg/mL,respectively,which are increased by 32 . 6%and 29 . 4%compared with that of the original formula.The results of the study are helpful to reduce the cost and improve the competitiveness of enterprises after being applied to large-scale production.
作者
随子华
孙术超
鞠加学
葛鹍鹏
赵国忠
陈瑶瑶
SUI Zihua;SUN Shuchao;JU Jiaxue;GE Kunpeng;ZHAO Guozhong;CHEN Yaoyao(Huasheng Company Limited of NCPC,Shijiazhuang,Hebei 052160,China;National Engineering Research Center of Microbial Medicine,Shijiazhuang,Hebei 052165,China;Hebei Industrial Microbial Metabolic Technology Innovation Center,Shijiazhuang,Hebei 052165,China;New Medicine Research and Development Company of NCPC,Shijiazhuang,Hebei 052165,China)
出处
《河北工业科技》
CAS
2023年第6期436-444,共9页
Hebei Journal of Industrial Science and Technology
关键词
发酵工程
两性霉素B
常压室温等离子体
紫外诱变
响应面筛选
fermentation engineering
amphotericin B
atmospheric and room temperature plasma(ARTP)
ultraviolet induction
response surface screening
