红曲色素高产菌株的高通量选育

High-throughput screening for high-yield strains of Monascus pigment

该文结合24深孔板微型化培养和酶标仪检测,建立了红曲色素高产菌株的高通量筛选方法,此法简单、快速、准确且自动化水平较高。确定最佳培养条件为装液量1.2mL/孔(每孔体积为10mL),接种量9%;找出红曲色素最大吸收峰值,确定酶标仪检测波长为530nm;通过紫外-氯化锂复合诱变处理,经过高通量筛选获得9株正突变株,其中菌株D39微孔板发酵色价达42.7U/mL,比出发菌株提高了46.2%;将D39分离纯化并用摇瓶验证,得到纯菌株D39-4的摇瓶发酵色价为206.5U/mL,比出发菌株提高了51.1%,具有很好的工业应用前景。

An economical and rapid high-throughput primary screening strategy was developed for high-aerobic Monascus purpureus cultivation.The good correlation of fermentation results with differing-scale cultivations confirmed the feasibility of utilizing the 24-deep microtiter plates（MTPs） as a scale-down tool instead of shake flasks for culturing Monascus pigment producing microbes.Supernatant obtained by a rapid centrifugation step after fermentation broth was extracted with ethanol,then it was evaluated by 96 deep-well and microplate reader at 530nm,which is more rapid and sensitive.Nine high mixture strains in the plate wells were obtained after compound mutagenesis using the high throughput screening method.The traditional dilution-plate method was employed to further isolate the high-yield single colony from the selected high-yield mixture strains.The pigment yield of D39-4 increased by 51.5%,and had stable inheritance after verification in the shake flask experiments.