棘白霉素B液体发酵培养的菌丝形态研究及发酵工艺优化  

Study on mycelia morphology of echinocandin B liquid fermentation and optimization of fermentation process development

作　　者：郝惠云 王耀耀[1] 朱研研 王亚莉[1] 李红珍 Hao Hui-yun;Wang Yao-yao;Zhu Yan-yan;Wang Ya-li;Li Hong-zhen(New Drug Research&Development Company of NCPC,National Engineering Research Center of Microbial Drugs,Shijiazhuang 052165)

机构地区：[1]华北制药集团新药研究开发有限责任公司,微生物药物国家工程研究中心,石家庄052165

出　　处：《中国抗生素杂志》2021年第12期1109-1114,共6页Chinese Journal of Antibiotics

摘　　要：目的明确棘白霉素B的菌丝形态特点,建立新的棘白霉素B发酵工艺。方法以构巢曲霉(Aspergillus nidulans)NCPC1246为出发菌株,发酵产生新型芬净类抗真菌药物阿尼芬净(anidulafungin)先导性化合物棘白霉素B(echinocandin B,ECB)。通过激光诱变和显微镜检,对菌球直径、菌丝形态变化规律和ECB产量进行相关性分析。依据ECB菌丝形态特点,优化发酵工艺。结果通过激光诱变筛选得到突变株NCPC1246-36,其菌球直径约为1.0~1.2mm,具备球形紧实内核。依据菌丝形态特点,调整摇瓶通气量参数和培养周期,确定八层纱布、转速220r/min和周期为168~192h。在50L发酵罐上尝试新的通气量和周期工艺,放罐单位达到(3354±47)μg/mL,较原工艺水平提高37.6%±0.9%。结论棘白霉素B发酵中,遵循菌丝形态特点,调整通气条件和培养周期,可以获得高产量。Objective The mycelium characteristics of echinomycin B were determined,and a new fermentation process of echinomycin B was established.Methods A starting strain of Aspergillus nidulans NCPC1246 was fermented to produce echinocandin B,which is the lead compound of new antifungal drug-anidulafungin.By laser mutagenesis and microscopic examination,correlation analysis was carried out on the hypha diameter,rule of hypha morphological change,and ECB yield.According to the morphological characteristics of mycelium,the fermentation process was optimized.Results The mutant strain NCPC1246-36 was obtained by laser mutagenesis,and the diameter of the fungus ball was 1.0~1.2mm,with a spherical compact core.According to the morphological characteristics of mycelium,the ventilation parameters and the culture period of shaking flask were adjusted,which were eight layers of gauze,the rotational speed of 220r/min,and the period of 168~192h.A new aeration process and culture cycle was amplified by secondary fermentation in 50L fermenters,and the titer of the releasing tank reached(3354±47)μg/mL,which was 37.6%±0.9%higher than the original level.Conclusion Following the morphological characteristics of hyphae,by adjusting ventilation conditions and culture cycle,the high yield of echinocandin B can be obtained.

关 键 词：棘白霉素B 构巢曲霉 激光诱变 菌丝球 二级发酵 动力条件 

分 类 号：R978.5[医药卫生—药品]