纳他霉素高产突变菌株高通量筛选方法的建立

为了提高纳他霉素（natamycin）高产菌株诱变后的筛选效率，建立一种24孔深孔板/酶标仪发酵初筛和摇瓶发酵复筛的高通量筛选检测方法，得到在24孔深孔板发酵结果与摇瓶发酵有很好的一致性，证实了酶标仪检测可以替代高效液相色谱法检测用于突变菌株的快速高通量筛选。以褐黄孢链霉菌（Streptomyces gilvosporeus TUST01）作为出发菌株，通过多轮的紫外诱变、常压室温等离子体（atmospheric and room temperature plasma,ARTP）以及硫酸二乙酯复合诱变、孔板初筛与摇瓶复筛，从887株突变株中筛选出遗传性稳定的高产菌株S. gilvosporeus Y-4-75，其摇瓶纳他霉素产量（5.95 g/L）与生物量（29.19 g/L）较出发菌株分别提高了31.93%和15.19%。

Establishment of a High-throughput Screening Method for High-yielding Mutant Strains of Natamycin

To improve the screening efficiency of the high - yielding mutant strains of natamycin，a high -throughput screening strategy was developed for the initial screening of 24-well deep-well plate/enzymatic assay（UV spectrophotometry）fermentation and the re-screening of shake flask（HPLC method）fermentation.The results obtained in the 24-well deep-well plate fermentation were in good agreement with that in the shake flask fermentation，confirming that the enzymatic assays can replace the HPLC assay for high - throughput screening of the mutant strains. With Streptomyces gilvosporeus TUST01 as the starting strain，high-yielding strain S. gilvosporeus Y-4-75 was selected from 887 mutant strains by multiple rounds of ultraviolet（UV），atmospheric and room temperature plasma（ARTP），diethyl sulfate（DES）mutagenesis，initial screening and rescreening of shake flask，and its natamycin yield in shake flasks（5.95 g/L）and biomass（29.19 g/L）were increased by 31.93% and 15.19%，respectively compared with those of the starting strain.