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红曲色素高产菌株的高通量选育
引用本文:石文娟,谭俊,储炬,郭元昕,庄英萍,张嗣良.红曲色素高产菌株的高通量选育[J].中国酿造,2012,31(7):25-28.
作者姓名:石文娟  谭俊  储炬  郭元昕  庄英萍  张嗣良
作者单位:华东理工大学生物反应器工程国家重点实验室,上海,200237
基金项目:国家重点基础研究发展计划‘973计划’(2012CB721006);国家高技术研究发展计划‘863计划’(2012AA021201)
摘    要: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.

关 键 词:红曲色素  微孔板  高通量  复合诱变  混合培养

High-throughput screening for high-yield strains of Monascus pigment
SHI Wenjuan , TAN Jun , CHU Ju , GUO Yuanxin , ZHUANG Yingping , ZHANG Siliang.High-throughput screening for high-yield strains of Monascus pigment[J].China Brewing,2012,31(7):25-28.
Authors:SHI Wenjuan  TAN Jun  CHU Ju  GUO Yuanxin  ZHUANG Yingping  ZHANG Siliang
Affiliation:(State Key Laboratory of Bioreactor Engineering,East China University of Science & Technology,Shanghai 200237,China)
Abstract: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.
Keywords:Monascus pigment  micro-bioreactor  high-throughput  compound mutation  mixture culture
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