利用Bacillus subtilis CW-116 發酵香菇柄生產抗氧化活性物質

Production of antioxidative activity compounds from shiitake stipes by Bacillus subtilis CW-116

葛俊宏(Ko Chun Hung)

指導教授 ： 張駿志 ； 黃晉修

亞洲大學/健康學院/保健營養生技學系碩士班/碩士(2008年)

摘要

本實驗主要利用Bacillus subtilis CW-116以香菇柄作為營養源生產抗氧化物質，並探討不同的培養條件對於抗氧化物質生產的影響。在抗氧化活性測定包含有清除DPPH自由基活性、螯合亞鐵離子能力、還原力、ABTS清除自由基活性(Trolox equivalent antioxidant capacity ,TEAC)，並探討最適合培養條件，其中包括最適培養時間、培養濃度及培養體積。進一步探討發酵液分離純化後其理化性質。發酵液進行聚焦層析和硫酸銨沉澱來分離蛋白質，並測定其抗氧化活性。由結果顯示香菇柄經B. subtilis CW-116 發酵培養後可有效提升其抗氧化活性。最適培養條件：培養時間三天、濃度4％、體積75mL有最高清除DPPH自由基活性為88.34±0.54 (trolox eq mg/g sample) (p<0.05)；培養時間二天、濃度6％、體積100mL之螯合亞鐵離子能力為63.91±0.56% (p<0.05)；培養時間五天、濃度4％、體積100mL之還原力為1.12±0.01 (p<0.05)；培養時間二天、濃度4％、體積100mL之ABTS清除自由基活性為390.45±0.53 (trolox eq mg/g sample) (p<0.05)。另外，本實驗結果發現發酵液中蛋白質及酚類化合物都具有抗氧化活性。綜合以上結果，香菇柄萃取物具有強力抗氧化活性，在天然抗氧化物製備上甚具潛力。

關鍵字

香菇柄 ； 抗氧化活性 ； 聚焦層析

並列摘要

The shiitake stipes was used in this project as a nutritional source for the production of antioxidants from Bacillus subtilis CW-116, and the effects of different culture conditions on antioxidant production were investigated. DPPH free radical scavenging activity, ferrous ion-chelating ability, reducing ability, and ABTS free radical scavenging activity (Trolox Equivalent Antioxidant Capacity assay, TEAC) were measured in order to evaluate antioxidant activity. The effects of different culture times, concentrations, and volumes were compared in order to obtain the optimal culture conditions. To further investigate the physical and chemical properties of the active components, the fermentation broth was further separated and purified using chromatofocusing and ammonium sulfate precipitation, and the isolated active components were then analyzed to determine the antioxidant activity. The results showed that the antioxidant activity of B. subtilis CW-116 was significantly enhanced when cultured with shiitake stipes. The optimal culture conditions for different antioxidant activities were as follows: at a culture time of three days, concentration of 4%, and volume of 75mL, the culture exhibited the highest DPPH free radical scavenging activity of 88.34±0.54 (trolox eq mg/g sample) (p<0.05); at a culture time of two days, concentration of 6%, and volume of 100mL, the culture exhibited the highest ferrous ion-chelating ability of 63.91±0.56% (p<0.05); at a culture time of five days, concentration of 4%, and volume of 100mL, the culture exhibited the highest reducing ability of 1.12±0.01 (p<0.05); and at a culture time of two days, concentration of 4%, and volume of 100mL, the culture exhibited the highest ABTS free radical scavenging activity of 90.45±0.53 (trolox eq mg/g sample) (p<0.05). In addition, it was demonstrated in this study that both the proteins and phenolic compounds in the fermentation broth exhibited antioxidant activity. In conclusion, extracts of the shiitake stipes have a strong antioxidant activity, and may be of great potential for use in the preparation of natural antioxidants.

並列關鍵字

shiitake stipes ； antioxidant activity ； chromatofocusing

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