吡咯喹啉醌菌株复合诱变及其高通量筛选

Compound Mutagenesis and High⁃throughput Screening of Pyrroloquinoline Quinone Strains

以一株能产生吡咯喹啉醌(pyrroloquinoline quinone,PQQ)的细菌为原始菌株,首先通过紫外、微波和氯化锂单一诱变,然后再进行紫外⁃微波⁃氯化锂三者复合诱变处理。研究结果表明,紫外照射50 s、微波(350 W)照射60 s和氯化锂浓度为0.4%处理效果最佳,获得突变株C5Y⁃6⁃4,该菌株摇瓶产量达134.9 mg/L,是原始菌株的1.32倍。通过96孔板高通量筛选和高效液相色谱法检测,结果表明,采用复合诱变在一定程度上提高了吡咯喹啉醌的产量,且突变株C5Y⁃6⁃4遗传稳定性好。

Starting with a bacterium capable of producing pyrroloquinoline quinone(PQQ),single mutagen⁃esis was initially performed using ultraviolet,microwave,and lithium chloride treatments individually,fol⁃lowed by a combined treatment of ultraviolet⁃microwave⁃lithium chloride.Research results showed that the op⁃timal conditions were achieved with ultraviolet irradiation for 50 s,microwave exposure(at 350 W)for 60 s,and 0.4%lithium chloride concentration,leading to the isolation of mutant strain C5Y⁃6⁃4,which exhibited a shake flask yield of 134.9 mg/L,1.32 times higher than the starting strain.High⁃throughput screening using a 96⁃well plate and analysis via high⁃performance liquid chromatography revealed that the compound mutagen⁃esis approach significantly increased the production of PQQ.Mutant strain C5Y⁃6⁃4 showed good genetic sta⁃bility.