臺灣博碩士論文加值系統

從雲林虎尾當地市場購得四季檸檬(Citruslimon (L.) osbeck)，而紅茶購自天仁茗茶之市售日月潭阿薩姆紅茶包，並以5%(w/v)含量在90oC熱水中萃取10min後得到紅茶萃取液(black tea water extract; BTWE)，漢氏醋桿菌(Komagataeibacter hansenii)是來自台灣當地康普茶菇菌所篩選出一種醋酸桿菌。在BTWE中加入10%(w/v)的檸檬與10%(w/v)蔗糖(定義為LsBT)作為K. hansenii的培養基質。另外，來自甘露醇基質(mannitol broth)前培養基質的K. hansenii被使用於探討有添加與無添加K. hansenii菌種發酵LsBT基質(分別相對定義為LsBTwKh與LsBTwoKh)之不同發酵天數(0-30 days)對LsBT發酵上清液之pH值、糖度(°Brix)及抗氧化活性探討。對LsBTwKh上清液pH值與糖度而言，其值會隨發酵時間增長而略微緩慢下降，在30天的發酵過程中LsBTwKh的pH值從第0天的pH 3.0緩慢下降到第30天的pH 2.5，而LsBTwoKh的pH值一直維持在pH3.0左右，而且在發酵期間LsBTwKh和 LsBTwoKh糖度(oBrix)均未發生明顯變化。對LsBT上清液之抗氧化活性定義為DPPH(1,1-diphenyl 2-picrylhydrazyl)自由基清除能力(scavenging activity)、總酚類化合物含量(total phenolic content; TPC)與還原能力(reducing power; RP)，一般而言，LsBTwKh上清液之抗氧化活性皆高於LsBTwoKh上清液之抗氧化活性，這歸因於前培養在甘露醇基質之起始菌(starter) K. hansenii所致。對LsBTwKh上清液而言，其DPPH自由基清除能力(定義為DPPH自由基清除率;DPPH %)在前9-12天發酵期間無顯著地略微上升，而後即逐漸緩慢下降，而LsBTwoKh之DPPH %值在儲存期間即開始逐漸下降。再者，LsBTwKh的TPC值在前3天發酵時間上升，而後無顯著地略微上升至第15天發酵時即開始逐漸下降，但LsBTwoKh之TPC值在前18天儲存期間無明顯變化，而後即開始逐漸下降。另外，LsBTwKh的RP值在前6天發酵時間上升，而後即開始逐漸下降，然而LsBTwoKh的RP值在儲存期間即開始逐漸下降。接著分別對LsBTwKh與LsBTwoKh上清液的DPPH%、TPC與RP值進行相互迴歸關係的探討，然而，對 LsBTwKh與LsBTwoKh上清液的TPC與RP值無法得到線性關係，但兩者的DPPH%-TPC與DPPH%-RP可以得到迴歸關係值DPPHmax%與½DPPHmax%濃度分別接近於86.93%與0.01963 mg GAE/mL，而DPPH%-RP之DPPHmax%與½DPPHmax%濃度值分別接近於82.64%與0.02994 mg Vit.C/mL，LsBTwKh的迴歸關係值皆優於LsBTwoKh者(分別相對接近於72.20%與0.02165 mg GAE/mL與83.37%與0.02699 mg Vit.C/mL，這些迴歸關係結果顯示應該歸因於K. hansenii起始菌前培養狀態所致，而且顯示要達到相同DPPH自由基清除能力時，RP濃度必須約1.5倍於TPC濃度，因此TPC貢獻較多。本研究結果顯示，K. hansenii起始培養於甘露醇基質可改進提高發酵檸檬紅茶上清液的抗氧化活性能力，且將貢獻於具有抗氧化活性的新健康型康普檸檬紅茶之研究發展。

Lemon (Citruslimon (L.) osbeck) was purchased from the local market in Huwei, Yunlin, and the black tea was purchased from the commercially available Sun Moon Lake Assam black tea bags from Tenren Tea, and was heated at 90oC with 5% (w/v) content. After extraction in water for 10 minutes, black tea extract (black tea water extract; BTWE) was obtained. Acetobacter hansenii (Komataeibacter hansenii) is an acetobacter isolated from the local kombucha mushroom in Taiwan. 10% (w/v) lemon and 10% (w/v) sucrose (defined as LsBT) were added to BTWE as the culture substrate for K. hansenii. In addition, K. hansenii from the pre-culture substrate of mannitol broth was used to investigate the different fermentation days (0- 30 days) to investigate the pH value, sugar content (°Brix) and antioxidant activity of LsBT fermentation supernatant. The pH value and sugar content of LsBTwKh supernatant will decrease slightly with the increase of fermentation time. During the 30-day fermentation process, the pH value of LsBTwKh slowly decreased from pH 3.0 on day 0 to pH 2.5 on day 30, while the pH value of LsBTwoKh remained around pH3.0, and neither LsBTwKh nor LsBTwoKh sugar content (oBrix) changed significantly during the fermentation. The antioxidant activity of LsBT supernatant was defined as DPPH (1,1-diphenyl 2-picrylhydrazyl) radical scavenging activity (scavenging activity), total phenolic compound content (total phenolic content; TPC) and reducing power (reducing power; RP). In general, the antioxidant activity of the LsBTwKh supernatant was higher than that of the LsBTwoKh supernatant, which was attributed to the starter K. hansenii pre-cultured on a mannitol matrix. For LsBTwKh supernatant, its DPPH free radical scavenging capacity (defined as DPPH free radical scavenging rate; DPPH %) did not significantly increase slightly during the first 9-12 days of fermentation, and then gradually decreased slowly, while the DPPH of LsBTwoKh. The % value begins to gradually decrease during storage. Furthermore, the TPC value of LsBTwKh increased during the first 3 days of fermentation, and then increased slightly without a significant increase until the 15th day of fermentation, and then began to gradually decrease, but the TPC value of LsBTwoKh did not change significantly during the first 18 days of storage, and then began to decrease. decreasing gradually. In addition, the RP value of LsBTwKh increased during the first 6 days of fermentation and then began to decrease gradually, whereas the RP value of LsBTwoKh began to decrease gradually during the storage period. Then, the mutual regression relationship between the DPPH%, TPC and RP values of LsBTwKh and LsBTwoKh supernatant was discussed. However, a linear relationship could not be obtained for the TPC and RP values of the supernatants of LsBTwKh and LsBTwoKh, but the regression relationship values of DPPH%-TPC and DPPH%-RP could be obtained. The DPPHmax% and ½DPPHmax% concentrations were close to 86.93% and 0.01963, respectively. mg GAE/mL, while the DPPHmax% and ½DPPHmax% concentrations of DPPH%-RP were close to 82.64% and 0.02994 mg Vit.C/mL, respectively. The regression relationship values of LsBTwKh were better than those of LsBTwoKh (relatively close to 72.20% and 0.02994 mg Vit.C/mL, respectively). 0.02165 mg GAE/mL and 83.37% and 0.02699 mg Vit.C/mL, these regression relationship results show that it should be attributed to the pre-culture state of K. hansenii starting bacteria, and show that to achieve the same DPPH free radical scavenging ability, The RP concentration must be about 1.5 times higher than the TPC concentration, so TPC contributes more. The results of this study show that K. hansenii initial culture on mannitol substrate can improve the antioxidant activity of fermented lemon black tea supernatant, and will contribute to Research and development of a new healthy type of Kombucha lemon black tea with antioxidant activity.

摘要......i
Abstract......iii
誌謝......v
目錄......vi
表目錄......viii
圖目錄......ix
符號說明......x
第一章 緒論......1
1.1前言......1
1.2研究目的......2
第二章 文獻回顧......3
2.1康普茶的發現......3
2.2康普茶的菌種......4
2.3康普茶的化學成份......6
2.4細菌纖維素......7
第三章 材料與方法......8
3.1 實驗材料......8
3.2 實驗藥品......8
3.3 實驗儀器......9
3.4 菌種的篩選......10
3.5 前培養......10
3.6 主培養......10
3.7 pH值與糖度測定......12
3.8 DPPH自由基清除能力測定......12
3.9總酚類化合物含量測定......13
3.10 總還原能力測定......14
第四章 結果與討論......15
4.1 pH值與糖度變化......15
4.4 DPPH 自由基去除能力測定......17
4.3 總酚類化合物含量測定......19
4.2 還原能力測定......21
4.5 還原能力、總酚類化合物含量與DPPH自由基去除能力關係......23
第五章 結論......27
參考文獻......28
附錄一......33
附錄二......34
附錄三......35
附錄四......36
附錄五......37
附錄六......38
附錄七......39
附錄八......40
Abstract......41

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