牛樟树水提物中促进樟芝无性产孢的效应物鉴定及其添加量优化

Identification of Water-Soluble Compounds from Cinnamomum kanehirae Hay Promoting the Asexual Sporulation of Antrodia cinnamomea and Optimization of Their Addition Levels in the Culture Medium

为明确牛樟树水提物(aqueous extract of Cinnamomum kanehirae Hay,CWE)中促进樟芝(Antrodia cinnamomea)深层发酵无性产孢的效应物。首先,采用醇沉法及不同有机溶剂分级萃取法对CWE进行分离,并以产孢量为检测指标,考察不同组分对樟芝无性产孢的影响。结果表明,30μg/mL的CWE醇沉上清氯仿萃取物LFE和50μg/mL的醇沉上清乙酸乙酯萃取物YZE对樟芝无性产孢有明显的促进效果,且LFE的促进效果明显优于YZE,表明效应物主要存在于LFE和YZE中。随后,采用液相色谱-质谱联用仪对CWE不同萃取组分进行成分分析,并结合不同组分对樟芝产孢的影响效果,确定主要效应物为赤藓糖醇。最后,考察赤藓糖醇(纯度98%)对樟芝深层发酵性能的影响,结果表明,赤藓糖醇对樟芝无性产孢有显著促进效果,且在最佳添加量(1.0μg/mL)下,产孢量较对照组提高55.17%;同时,1.0μg/mL赤藓糖醇显著促进樟芝深层发酵菌丝体生长及胞内多糖的合成,生物量和胞内多糖产量较对照组分别提高18.65%和260.13%,但对三萜的合成无显著影响。

In order to determine compounds present in the aqueous extract of Cinnamomum kanehirae Hay(CWE)that promote the asexual sporulation of Antrodia cinnamomea in submerged fermentation.First,CWE was isolated by alcohol precipitation and fractional extraction with different organic solvents.The influence of the obtained fractions on the asexual sporulation of A.cinnamomea was investigated.It was showed that the chloroform(at 30μg/mL)and ethyl acetate(at 50μg/mL)extracts of the supernatant after ethanol precipitation of CWE presented remarkable promoting effects on the sporulation of A.cinnamomea,and the effect of LFE was significantly more pronounced than that of YZE,indicating that both LFE and YZE contained compounds that promote the sporulation of A.cinnamomea.Subsequently,liquid chromatography-mass spectrometry(LC-MS)was used to analyze the chemical components of the fractions obtained from CWE,and erythritol was considered as the major component that promotes the sporulation of A.cinnamomea.Finally,the effect of erythritol with a purity of 98%on the fermentation performance of A.cinnamomead was investigated.The result showed that erythritol did significantly promote the sporulation of A.cinnamomea and increased the spore production by 55.17%compared with the control group at the optimal concentration of 1.0μg/mL.Meanwhile,1.0μg/mL erythritol significantly promoted the mycelial growth and synthesis of intracellular polysaccharides(IPS)of A.cinnamomea in submerged fermentation and increased the biomass and the yield of IPS by 18.65%and 260.13%,respectively.However,erythritol had no significant effect on the synthesis of triterpenes in A.cinnamomea.