The invention discloses a method for preparing succinic acid by fermentation of sucrose, including strain activation, seed cultivation, anaerobic fermentation of three steps, and creatively in the fermentation step using sucrose and sucrose glucose mixed with sugar as carbon source. The technical scheme of the invention of the succinic acid yield and production rate using glucose fermentation results in high sucrose concentration was 100g/L, culture can produce 57.1g/L succinic acid in serum in anaerobic fermentation tank, the fed batch fermentation can produce single 60.5g/L Ding two acid. The best ratio of sucrose and glucose in sucrose glucose mixed sugar was 4:1, and the concentration and yield of succinic acid reached the maximum, respectively, 61.6g/L, 85.3%.
【技术实现步骤摘要】
【技术保护点】
一种利用蔗糖发酵制备丁二酸的方法,其特征在于,所述方法包括如下步骤:(1)斜面培养:将冻存菌种融化并在斜面培养基中进行平板划线后,在温度为30~40℃的厌氧培养箱中活化培养24~72?h,活化好的单菌落用于种子培养基接种和菌株保存;(2)种子培养:向100?mL血清瓶中加入种子培养基30~60?mL,115~121?℃灭菌15~25?min,冷却后接入斜面培养菌株,通入CO2?1~3?min,30~40℃培养,摇床转速100~200?r/min,培养时间为10~16?h,用于发酵培养基接种;(3)血清瓶培养:100?mL?血清瓶中加入发酵培养基20~50?mL,加入碱性中和剂,115~121℃灭菌15~25?min,冷却后接入种子培养液,通入CO2?1~3min保证厌氧环境,接种完成后,培养温度为30~40℃,摇床转速100~200?r/min,培养时间为24~72?h;(4)3L发酵罐发酵培养:3?L发酵罐中发酵培养基体积为1.5~2?L,接种量为5~10%(v/v),温度为30~40?℃,发酵罐中持续通入CO2以保持发酵体系的厌氧环境,搅拌转速100~300?rpm,发酵过程分别采 ...
【技术特征摘要】
【专利技术属性】
技术研发人员:姜岷,戴文宇,奚永兰,徐蓉,张九花,陈可泉,张敏,
申请(专利权)人:南京工业大学,
类型:发明
国别省市:
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