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  • 學位論文

舞菇固態與液體培養條件之探討及其發酵液之抗氧化活性分析

Solid-state and Submerged Cultivation and Fermentation Fluid Antioxidant Activity of Grifola frondosa

指導教授 : 施養佳 張竣維
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摘要


舞菇(Grifola frondosa)是一種食藥兼用的真菌,經研究發現舞菇具有抗腫瘤、強化免疫系統、降低血糖、血壓和膽固醇、抗糖尿病及瘦身等效果,所以極受國際生物與醫學界之重視。一般傳統栽培方式由接種至生成子實體需耗費很長的時間,而運用發酵技術生產菌絲體和多醣體,不僅能縮短培養時間,且品質和產量都較穩定。有鑑於此,本研究之目的有三項: 首先探討五種品系之舞菇其最佳碳源及氮源的固態培養條件;接著探討舞菇深層培養方式調控轉速與初始培養pH值對其菌絲體、粗多醣和培養後pH值的變化;最後分析舞菇發酵液之總多酚與類黃酮含量,以及發酵液的清除DPPH自由基能力。第一部分的實驗結果顯示,舞菇菌絲體培養20天後,其生長情形最佳的碳源與氮源條件如下:以果糖4%為碳源與玉米漿2%為氮源,使舞菇品系T1生長達40.17 mm;葡萄糖3%及玉米漿1.5%,讓舞菇品系T2生長達42 mm;果糖4%和酵母萃取物1.5%,促使舞菇品系BT生長達42.83 mm;葡萄糖4%和玉米漿1.5%,使舞菇品系D-4生長達41 mm;葡萄糖4%和玉米漿1.5%,使舞菇品系5-T生長達42 mm。第二部分之實驗目的是探討不同轉速與初始pH值對舞菇深層培養之影響。經由實驗結果發現,所有品系(T1、T2、BT、D-4、5-T)在rpm 150的培養下,粗多醣的產量最高,分別為2.82±0.56、2.11±0.13、3.24±0.71、2.85±0.39、2.06±1.23 g/L,但對菌絲體和最終pH值的變化無顯著影響。初始pH值部分,舞菇品系T1、T2、BT,分別在初始pH值為5.5時,有最高菌絲產量1.83 g/L、4.18 g/L、3.16 g/L,而舞菇品系D-4和5-T其最高菌絲體產量分別在初始pH值6.5(3.83 g/L)和4.5(3.14 g/L)。此外,舞菇粗多醣之產量部分,T1、T2、BT和5-T,分別在初始pH值為4.5時,最高粗多醣產量分別為8.39 g/L、7.57 g/L、5.56 g/L和5.07 g/L。而D-4則是在初始pH值6.5時,粗多醣產量最高為7.07 g/L。第三部分抗氧化成分之分析結果顯示,以T2在果糖3%和玉米漿1.5%且初始pH值為6.5時,其發酵液的總多酚和類黃酮成分為所有品系中最高,分別為29.75 mg/g及1.20 mg/g。而清除DPPH自由基能力部分,T1、T2、BT、D-4和5-T清除DPPH自由基能力最高分別為53.70%、46.88%、57.81%、48.68% 及55.54%。以EC50來看,5-T 品系之舞菇以4%葡萄糖和1.5%玉米漿進行深層培養,且初始pH值為4.5之發酵液的抗氧化效果最佳,其EC50為23.67 mg/mL。總結上述之研究結果,舞菇菌絲體在碳源方面較適合用較高濃度的(4~3%)葡萄糖和果糖來培養,而氮源部份,每個品系皆適用於較低濃度玉米漿(2~1.5%),調控初始pH值(弱酸的情形下)對菌絲體和粗多醣產量沒有顯著影響,但對抗氧化能力會有些微影響。未來可以再深入探討不同種類的碳氮源對於舞菇之抗氧化力的影響,以及探討添加不同界面活性劑或是植物油等,對舞菇深層培養菌絲體及多醣的產量與抗氧化能力之影響。本論文的成果,不僅可提供菇農對於不同品系舞菇之栽培及深層培養的相關資訊,未來亦可將舞菇發酵液應用於機能性保健食品的開發,以增加舞菇之產業利用性及提升其品之附加價值。

並列摘要


Grifola frondosa is one kind of medical and edible fungus, and it has been found lots of functions such as: anti-tumor, strengthen the immune system, lowering blood glucose, blood pressure and cholesterol, and weight-loss and other anti-diabetic effects. The international biological and medical community has been highly paid attention to G. frondosa recently. Due to the traditional cultivation of G. frondosa from the inoculation to produce fruiting bodies which takes a long time, the fermentation technology to produce mycelium and polysaccharides can not only shorten the training time, but also increase the quality and the yield stably. For the above reasons, the three purposes of this study are as follows. (1) To investigate the best carbon and nitrogen source of the five strains of G. frondosa by solid-state culture. (2) To study the production effects of G. frondosa mycelium and polysaccharide by different rpm and different initial pH value of submerged culture. (3) To analyze total polyphenol and flavonoid content and fermentation DPPH radical scavenging ability of the fermentation broth of G. frondosa. The first part of experimental results showed that best carbon source and the nitrogen source for the mycelium growth of five strains G. frondosa after 20 days cultivation were as follows: the 4% fructose as the carbon source and the 2% corn steep powder as the nitrogen source caused the mycelium growth of T1 strain reached 40.17 mm; the 3% glucose and the 1.5% corn steep powder let the mycelium growth of T2 strain reached 42 mm; the 4% fructose and the 1.5% yeast extract urged the mycelium growth of BT strain reached 42.83 mm; the 4% glucose and the 1.5% corn steep powder caused the mycelium growth of D-4 strain reached 41 mm; the 4% glucose and the 1.5% corn steep powder caused the mycelium growth of 5-T strain reached 42 mm. The second part of the study was designed to investigate the different rpm and initial pH value on the impact of submerged culture of G. frondosa. According to the experimental results, All strains (T1, T2, BT, D-4 ,5-T) in rpm 150 under cultivation, the highest yield of polysaccharide were 2.82±0.56, 2.11±0.13, 3.24± 0.71, 2.85±0.39, 2.06±1.23 g/L, but the mycelium and the final pH of no significant impact of the change. Part of the initial pH value, T1, T2 and BT strains of G. frondosa, produced respectively the highest mycelial yields as 1.83 g/L, 4.18 g/L and 3.16 g/L when the initial pH value was 5.5. In addition, D- 4 and 5-T strains, produced the highest mycelial yields when the initial pH values were 6.5 (3.83 g/L) and 4.5 (3.14 g/L). Furthermore, the maximum polysaccharide production of G. frondosa T1, T2, BT and 5-T strains were 8.39, 7.57, 5.56 and 5.07 g/L when the initial pH was 4.5. The D-4 strain presented the highest yield of polysaccharides (7.07 g/L) when the initial pH was 6.5. The third part of the antioxidant ability experiments indicated that the fermentation broth of T2 strain contained the highest total polyphenols (29.75 mg/g) and flavonoids (1.20 mg/g) above other strains under the cultivation conditions of 3% fructose and 1.5% corn steep powder at initial pH of 6.5. Moreover, T1, T2, BT, D-4, and 5-T strains presented the highest ability of scavenging DPPH radicals were 53.70%, 46.88%, 57.81%, 48.68% and 55.54%. Especially, G. frondosa 5-T strain presented the best results of EC50 within five strains was 23.67 mg/mL under the fermentation conditions of 4% glucose, 1.5% corn steep powder and initial pH of 4.5. Summarize the above results, higher concentrations (4~3%) of glucose and fructose as the carbon resources were suitable for the mycelia growth of all five strains of G. frondosa. Lower concentrations of corn steep powder (2~1.5%) as nitrogen resources were suitable for all strains of G. frondosa. The initial pH values (the case of weak acid) of G. frondosa fermentation were not significantly affected the mycelium and polysaccharide productions, but they had a little impact on the antioxidant capacity of G. frondosa. The relationship between different carbon-nitrogen resources and antioxidant abilities; or between different surfactants such as vegetable oil and polysaccharide production of G. frondosa can be further investigated. Based on the achievement of this study, the related information can be provided to the farmers for the solid-state of cultivation and the submerged culture in five different strains of G. frondosa. In the future, the fermentation fluid of G. frondosa can be applied for the development of functional health products to increases the industrial utilities and to promote the additional values of G. frondosa.

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