Submerged fermentation production and characterization of intracellular triterpenoids from Ganoderma lucidum using HPLC-ESI-MS

利用液体发酵提高灵芝三萜产量及HPLC-ESI-MS鉴定灵芝菌丝中灵芝酸成分(英文)

As the main bioactive metabolites of Ganoderma lucidum, triterpenoids have various pharmacological effects. In this paper, the nutritional requirements and culture conditions of a submerged culture of G. lucidum were optimized using the response surface methodology; maximum mycelia biomass and intracellular triterpenoid production reached 1.87 g/100 ml and 93.21 mg/100 ml, respectively, for a culture consisting of wort 4.10% （0.041 g/ml） and yeast extract 1.89% （0.0189 g/ml）, pH 5.40. For the first time, we established that wort, which is cheap and abundant, can replace the more commonly used glucose as the sole source of carbohydrate. Using high-performance liquid chromatography-electrospray ionization-mass spectrometry （HPLC-ESI-MS）, 10 major ganoderic acids were tentatively identified based on the predominant fragmentation pathways with the elimination of H20 and CO2, as well as cleavage of the D-ring.

目的:通过优化液体发酵培养基提高灵芝三萜产量,并对其三萜成分进行初步鉴定。创新点:麦芽作为啤酒生产原料,含有丰富的糖类及其他营养物质,并且价格低廉,来源广泛。本文首次将麦芽汁作为碳源物质以提高灵芝三萜产量,并对其所含灵芝酸成分进行初步鉴定。方法:以灵芝菌丝生物量和胞内三萜产量为检测指标,对其液体发酵培养基中的碳(C)源和氮(N)源进行筛选,并利用单因素实验确定其相应浓度,再经响应面法优化液体发酵培养基(C源、N源、pH)(表1)。另一方面,酸碱转化法纯化灵芝三萜提取物后,利用高效液相色谱-电喷雾-质谱(HPLC-ESI-MS)法初步鉴定灵芝所含灵芝酸成分(表2)。结论:本实验响应面结果显示:当培养基含麦芽汁4.1%,酵母浸出粉1.89%,pH为5.40时,灵芝液体发酵菌丝生物量及胞内三萜产量均达到最优,分别为1.87g/100ml和93.21mg/100ml。经HPLCESI-MS鉴定,初步确定其含有10种灵芝酸成分。