荧光物质(MFA、MFB)的分离、结构鉴定及金华地区高产菌株的筛选

Isolation,Structure Characterization of Fluorescent Substances(MFA,MFB)and Screening of High-Yielding Strains in Jinhua Region

采用硅胶柱层析及制备型液相色谱仪对红曲米中两种荧光物质进行分离纯化,使用高效液相色谱法(HPLC)检测荧光物质纯度,然后使用高分辨质谱(ESI-HRMS)对两种荧光物质进行分析,得到两种荧光物质的分子量分别为356和384,ESI-MS/MS二级质谱把两者鉴定为monasfluore A(MFA)和monasfluore B(MFB);从金华地区红曲米中分离得到10株红曲菌株,经固态发酵采用HPLC法分析,筛选获得1株高产MFA、MFB的菌株WZWZ,该菌株发酵制得红曲米中MFA含量为3.63 g/kg,MFB含量为7.29 g/kg,对WZWZ菌株进行外观形态学及显微观察、ITS基因序列测定与分析,最终将菌株WZWZ鉴定为紫色红曲霉(Monascus purpureus)。

In this experiment,two kinds of fluorescent substances in red yeast rice or red kojic rice were first separated and purified by silica gel column chromatography and preparative liquid chromatography,then the purity of the fluorescent substance was detected by high performance liquid chromatography(HPLC),followed by analysis with high resolution mass spectrometry(ESI-HRMS)for the two fluorescent substances,and obtained molecular weights of the two fluorescent substances were at 356 and 384 respectively,and identified the two by ESI-MS/MS secondary mass spectrometry as monasfluore A(MFA)and monasfluore B(MFB);Ten Monascus strains of were isolated from red yeast rice in Jinhua region.A strain of high-yield MFA and MFB WZWZ was obtained by solid state fermentation.The MFA content of red yeast rice was at 3.63 g/kg and the MFB content was at 7.29 g/kg,the appearance morphology of WZWZ strain was observed under microscope,ITS gene sequence was determined and analyzed,finally identified strain WZWZ as Monascus purpureus.