Abstract: To improve the microbial fermentation yield of ansamitocinP-3 (AP-3), 96-well plate-microplate high-throughput screening method was established by UV-spectrophotometry. UV mutation breeding was carried out using the original strain (Actinosynnema pretiosum ssp. auranticum). A mutant strain B24-13 was obtained, and the higher production of AP-3 was 2.03-fold (reaching 112.5mg/L) compared with that original strain after 7 days fermentation, and the fermentation medium was optimized by culture medium optimization. After the analysis of the experimental results, the optimal medium for the mutant strain B24-13 was sucrose 25g/L, glycerol 1.5g/L, corn steep liquor 25g/L, CaCO₃ 7g/L, isobutanol 2g/L, valine 0.5g/L, MgSO₄ ·7H₂O, 0.5g/L, and FeSO₄·7H₂O 0.01g/L. Under those conditions, the yield of AP-3 was (127.5±6.3)mg/L after 7 days fermentation. The yield of AP-3 can be further improved by the mutagenesis breeding and the optimization of the culture medium. It is proved that the UV-spectrophotometry method is feasible for the rapid screening AP-3 high production strain.

Key words: ansamitocinP-3, UV mutagenesis, UV spectrophotometry, culture medium optimization, orthogonal design

摘要： 为了提高安丝菌素P-3（AP-3）的发酵产量，通过紫外诱变并运用紫外-分光光度法建立96孔板高通量快速筛选体系对原始菌株（Actinosynnema pretiosum ssp.auranticum）进行育种，筛选获得了一株产量较高的突变菌株B24-13。发酵7天后，其发酵液中AP-3的含量为112.5mg/L，是原始菌株的2.03倍。随后通过单因素优化与正交实验设计对突变菌株B24-13进行发酵培养基优化，得到最优发酵培养基组分：蔗糖25g/L，甘油15g/L，玉米浆25g/L，CaCO₃ 7g/L，异丁醇2g/L，缬氨酸0.5g/L，MgSO₄·7H₂O 0.5g/L，FeSO₄·7H₂O 0.01g/L。对优化结果进行验证实验，发酵7天后AP-3的产量为（127.5±6.3） mg/L。实验结果表明：通过对生产菌株的诱变育种与发酵培养基优化可有效的提高AP-3的发酵产量，也从侧面验证了该研究所建立的96孔板高通量快速筛选体系用于AP-3高产菌株的初筛是可行的。

关键词: 安丝菌素P-3, 紫外诱变, 紫外-分光光度法, 培养基优化, 正交设计