油酸促进灵芝三萜液态深层发酵的工艺研究及规模化验证

Synthesis of triterpenes in liquid submerged fermentation of Ganoderma lingzhi promoted by oleic acid

灵芝三萜是灵芝中主要的活性成分之一,前期研究发现油酸可以促进灵芝三萜液态深层发酵下的发酵合成。本研究主要对油酸促进灵芝三萜液态深层发酵的工艺进行优化,并进行3L发酵罐规模的验证。通过单因素实验考察油酸的添加方式、添加时间和添加浓度对灵芝三萜的影响,结合响应面实验,获得最优工艺条件并进行验证:在发酵第32h添加1.21%高温灭菌油酸,最高灵芝三萜含量为42.69mg/g;在发酵第7h添加1.35%过滤除菌油酸,最高三萜含量为43.38mg/g,分别比对照提高2.04倍和2.08倍。在1000m L摇瓶中添加高温灭菌油酸和过滤除菌油酸,灵芝三萜含量分别为32.18和32.48mg/g,为对照的1.96倍和1.95倍;在3L发酵罐规模下灵芝三萜含量分别为28.66和25.13mg/g,为对照的1.62倍和1.42倍。本研究系统优化了油酸促进灵芝三萜液态深层发酵的工艺条件,并在与工业生产相对应的3L发酵罐上进行验证。该研究可为灵芝三萜的规模化发酵提供重要参考和借鉴。

Triterpene is one of the most important active ingredients in Ganoderma lingzhi.Previous studies found that oleic acid could promote the biosynthesis of triterpenes in liquid submerged fermentation of Ganoderma lingzhi.In this study,the process of liquid submerged fermentation for synthesis of triterpenes by using oleic acid as promoter was optimized and verified on the scale of a 3 L fermenter.The effects of the additive method,time for addition and concentration of oleic acid on triterpene synthesis were investigated by single factor experiment.The optimal process conditions were obtained and verified by response surface experiment.The highest triterpene content reaching 42.69 mg/g was obtained by adding 1.21%moist heat sterilized oleic acid at 32 h of fermentation,and by adding 1.35%filtered sterilized oleic acid at the 7 h of fermentation,the highest triterpene content was 43.38 mg/g,being 2.04 times and 2.08 times higher than triterpene content of the control,respectively.The triterpenes content was 32.18 mg/g and 32.48 mg/g respectively by adding moist heat sterilized and filterer sterilized oleic acid at 1000 m L scale of shake culture,being 1.96 times and 1.95 times higher as compared with the control.The triterpene content in 3 L fermenter was 28.66 mg/g and 25.13 mg/g,1.62 times and 1.42 times higher than that of the control.The technological conditions of liquid submerged fermentation of triterpenes promoted by oleic acid were successfully optimized and verified in a 3 L fermenter,corresponding to industrial production.This study provide an important reference for large-scale fermentation of Ganoderma lingzhi for production of triterpenes.