| Abstract: | 麩胱甘? (glutathione) 和麥角硫因 (ergothioneine) 具有抗氧化功效，屬於人體重要的抗氧化劑。由於麥角硫因無法在人體內合成，僅能由飲食中攝取，近幾年，學者發現食藥用菇中含有豐富的麩胱甘?與麥角硫因，可清除自由基及過氧化物質生成，達到有效預防老化、阿茲海默症及心血管疾病發生，並富含多種生理活性物質，如麥角固醇、維生素D2、三?類及多醣體等。本研究分別篩選最適子實體與菌絲體之高麩胱甘?與高麥角硫因含量。在食藥用菇子實體方面，以狼尾草杏鮑菇與柳松菇為最適條件，並探討冷藏試驗及栽培條件。狼尾草杏鮑菇與柳松菇，由冷藏試驗可知，保存越久麥角硫因含量會提升，麩胱甘?含量則是相反；杏鮑菇栽培介質與部位方面，麩胱甘?含量以狼尾草杏鮑菇 (2.23 mg/g dw) 最高，麥角硫因含量以牧草杏鮑菇菇頭 (2.40 mg/g dw) 最高；栽培天數方面，狼尾草杏鮑菇栽培 90 天有最高麩胱甘? (3.51 mg/g dw) 含量和麥角硫因 (4.25 mg/g dw) 含量；杏鮑菇菌傘變化以菌傘開之狼尾草杏鮑菇菇頭有較高含量之麩胱甘? (3.51 mg/g dw) 和麥角硫因 (2.97 mg/g dw)，結果得知，以狼尾草杏鮑菇菌傘越開與栽培 90 天之麩胱甘?及麥角硫因含量較佳。食藥用菇菌絲體方面，麩胱甘?與麥角硫因含量分別以舞菇 (3.82 mg/g dw) 和香杉芝 (2.58 mg/g dw) 最高，由於學者較少發現香杉芝含有較高麥角硫因含量，因此以香杉芝進行液態發酵實驗；香杉芝調整初始pH 值8 能提升麩胱甘? (3.46 mg/g dw) 含量；香杉芝添加4 mM與 8 mM 前驅物 (半胱胺酸、甲硫胺酸及組胺酸)，以第 7 天添加 4 mM半胱胺酸 (7day4Cys) 有最高麩胱甘? (1.98 mg/g dw) 及麥角硫因 (4.47 mg/g dw) 含量，第 11 天添加 8 mM 半胱胺酸 (11day8Cys) 有較高麩胱甘? (4.92 mg/g dw)；添加甲硫胺酸與第 7 天添加 4 mM 組胺酸能小幅度增加麩胱甘?含量，但麥角硫因含量無提升之效。因此，以 7day4Cys、11day8Cys 與香杉芝液態發酵 31天為最適含有麩胱甘?及麥角硫因 (AS) 進行生理活性成分、生理活性物質及生理活性成分比較。生理活性成分方面，三?含量以 11day8Cys (28.28 mg/g) > 7day4Cys (25.66 mg/g) > AS (12.72 mg/g)，多醣以 11day8Cys (13.86 mg/g) > 7day4Cys (12.61 mg/g) > AS (12 mg/g)，麥角固醇含量以 11day8Cys (3.23 mg/g) > AS (3.02 mg/g) > 7day4Cys (2.82 mg/g)，由結果可知，11day8Cys 與 7day4Cys 可提升三?、多醣含量；麥角固醇方面，11day8Cys 較無提升效果。因此推測，三?與麥角硫因屬於次級代謝產物，香杉芝液態發酵培養 31 天 (AS) 有較高麥角硫因和三?含量，另外發現添加半胱胺酸能提升三?及多醣含量，但 7day4Cys 麥角固醇含量降低，推測菌絲生長情形較差。抗氧化性質 EC50 值方面，7day4Cys 與 11day8Cys 皆提升清除 ABTS+ 自由基能力、清除 DPPH 自由基能力及還原力，與 AS 相比，總酚與類黃酮含量也提升，實驗結果表示，香杉芝添加半胱胺酸不僅能提升麩胱甘?含量、麥角硫因含量，同時也增加生理活性成分、抗氧化物質及抗氧化成分，因此可利用香杉芝以液態發酵添加半胱胺酸以提升抗氧化效果，未來能應用於保健食品及保養品。
Glutathione and ergothioneine act as the important antioxidants in human body. Since ergothioneine cannot be synthesized by the body, people can only absorb it by daily diet. Recently, scholars found that edible and medicinal mushrooms are rich in glutathione and ergothioneine which neutralize free radicals and effectively prevent aging, Alzheimer's disease and cardiovascular disease. In addition, the mushrooms are full of bioactive materials, such as ergosterol, vitamin D, triterpenes and polysaccharides. In the present study, we individually selected the optimal content of glutathione and ergothioneine from fruiting bodies and mycelia. In the experiment of edible and medicinal mushrooms, Pleurotus eryngii and Agrocybe aegerita are the optimal conditions to do the refrigerated test and to be cultivated. It can be seen from the refrigeration experiment of Pleurotus eryngii and Agrocybe cylindracea that the longer they keep, the contents of ergothioneine are increasing while glutathione are decreasing. In the experiment of Pleurotus eryngii cultivated by the medium and its part, Pleurotus eryngii contained the highest content of glutathione with 2.23 mg/g dw; In the experiment of different planting days,, Pleurotus eryngii planting 90 days contained the highest content of glutathione (3.51 mg/g dw) and ergotathione (4.25 mg/g dw); Changes of caps for Pleurotus eryngii contained the highest content of glutathione (3.51 mg/g dw) and ergotathione (2.97 mg/g dw). As a whole, changes of caps and 90 days were the best planting cycle to cultivate Pleurotus eryngii.In the experiment of edible and medicinal mushrooms mycelia, Grifola frondosa and Antrodia salmonea contained the highest content of glutathione (3.82 mg/g dw) and ergotathione (2.58 mg/g dw) respectively. It was hardly for scholars to discover the highest content of glutathione and ergothioneine in Antrodia salmonea, therefore, it was experimented by submerged fermentation as the basic concept ; the contents of glutathione of Antrodia salmonea for initial pH 8 value are increasing at submerged fermentation; Antrodia salmonea adding 4 mM and 8 mM precursor (cysteine、methionine and histidine), 7day4Cys contained the highest content of glutathione (1.98 mg/g dw) and ergotathione (4.47 mg/g dw). 11day8Cys contained the highest content of glutathione (4.92 mg/g dw); The addition of methionine can slightly increase the content of glutathione, but the content of ergotathione has no effect on it; Antrodia salmonea adding histidine can inhibit the content of glutathione and thioneine. Therefore, 7day4Cys, 11day8Cys and the submerged fermentation of Antrodia salmonea with high glutathione and ergotathione on 31th day (AS).In the aspect of physiologically active ingredients, the content of triterpenoids from high to low is in the sequence of 11day8Cys, 7day4Cys, AS (28.28 mg/g > 25.66 mg/g >12.72 mg/g); the contents of polysaccharide from high to low is in the sequence of 11day8Cys, 7day4Cys, AS (13.86 mg/g > 12.61 mg/g >12 mg/g); the contents of ergosterol from high to low is in the sequence of 11day8Cys, AS, 7day4Cys (3.23 mg/g > 3.02 mg/g > 2.82 mg/g). As the result, 11day8Cys and 7day4Cys can increase the contents of triterpenoid and polysaccharide, while 11day8Cys has no effect on ergosterol. According to the experimental results, triterpenoid and ergotathione are secondary metabolites, Antrodia salmonea contained the highest contents of glutathione and triterpenoid. In addition, it is found that the efficiency of triterpenoid and polysaccharide are increasing by adding cysteine, while the contents of ergosterol are decreasing for 7day4Cys which indicate that the growth of biomass is poor.In terms of antioxidant activity EC50 value substance, both of 7day4Cys and 11day8Cys increase the content of ABTS+ free-radicals scavenging ability, DPPH free-radicals scavenging ability and reducing power. To compare with AS, the total phenol and flavonoids have enhancements. To observe from the experiment, Antrodia salmonea with cysteine can not only increase glutathione and ergotathione but also enhance physiological activity ingredients, antioxidant propertiex and antioxidant components. Therefore, cysteine can be added in Antrodia salmonea by submerged fermentation to improve antioxidant activity. Moreover, it is positive to apply this concept on dietary supplements and facial care products in the future. |
