基質強化對提升傳統紅茶菇發酵纖維膜、多酚類及抗氧化活性之影響

孫慈霙

doi:10.6845/NCHU.2015.00062

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基質強化對提升傳統紅茶菇發酵纖維膜、多酚類及抗氧化活性之影響

Effects of substrate fortification on enhancing cellulosic pellicle, phenolic compounds and antioxidant activities during traditional kombucha fermentation

孫慈霙 , Ph.D　　Advisor：陳錦樹　　

英文 DOI： 10.6845/NCHU.2015.00062

紅茶菇；酚酸；纖維素；抗氧化；氧自由基吸收能力； Kombucha ； Phenolic compounds ； Cellulose ； Antioxidant activity ； Oxygen radical absorbance capacity

Abstract │ Reference (129) │ Altmetrics

Abstract 〈TOP〉

紅茶菇(kombucha、tea fungus)是一種以含糖紅茶液為基質經茶菇菌(醋酸菌與酵母菌)共同發酵產生特殊風味之傳統發酵飲品，由上層纖維薄膜及下層發酵液組成。目前紅茶菇極少研究是從強化基質作為探討，本研究主要目的為探討強化紅茶菇之發酵基質對提升紅茶菇發酵時纖維膜、多酚類及抗氧化活性之影響。
（一）本研究將不同劑量之咖啡因添加於含糖紅茶湯液(0~0.4 %, w/v)，接種主由酵母菌(Dekkera bruxellensis)和醋酸菌(Gluconacetobacter rhaeticus 及 Gluconobacter roseus)所組成之菌酛，接種量為20% (v/v)，次於29±1℃下靜置培養12天，總共分成5組(A, B, C, D, E)，探討對發酵液中酵母菌及醋酸菌菌數、pH、細菌纖維膜之產量、抗氧化、糖類及主要有機酸變化之影響。結果顯示，各組酵母菌及醋酸菌分別於發酵至第6天及第3天達最大菌量，分別為7.2 log (cfu/mL)及7.3 log (cfu/mL)。隨著發酵時間增加，pH由起始3.9逐漸下降至約2.8。在細菌纖維膜之產量上，發酵至12天添加0.4 %咖啡因的E組比未添加之傳統紅茶菇增加了26.5 %。蔗糖含量皆隨發酵時間增加而緩慢下降。葡萄糖於發酵初期下降迅速，但隨著咖啡因添加量增加，而消耗速率減緩。D及E組之果糖含量於發酵初期有累積的情況。有機酸含量皆隨發酵時間持續累積，醋酸含量以A組累積量最高，發酵至第12天約為17.5 g/L。E組之葡萄糖酸含量則於第3天具有最高濃度15.4 g/L。添加咖啡因可提供菌體生長額外的碳源，並促進發酵液中葡萄糖酸含量的累積，進而改變發酵液中有機酸含量之分布。D組發酵到第3天，C組發酵到第6天，DPPH自由基清除率達最大值。添加咖啡因共同發酵而得的改良式紅茶菇比傳統紅茶菇更具有較高之DPPH自由基清除能力及總抗氧化力。 (二) 本單元研究探討添加不同劑量之兒茶素(0~0.3 %, w/v)混合發酵培養成更具保健能力之飲品。探討發酵期間菌數、pH值、抗氧化特性之變化與糖類及有機酸含量。結果顯示，於紅茶液中添加兒茶素共同發酵之最終狀態，各組pH值雖然有變化上的差異，但發酵過程中之變化趨勢相似。添加兒茶素之各組菌量雖無顯著差異，卻可使發酵液代謝作用更加活化及促使代謝產物如有機酸的產生。DPPH自由基清除能力及總抗氧化能力皆以添加0.1%兒茶素組，發酵6天顯著高於傳統紅茶菇(p<0.05) ，添加兒茶素共同發酵而得的改良式紅茶菇比傳統紅茶菇具有較高且穩定之抗氧化力。發酵液之成分方面，蔗糖隨著發酵天數逐漸降低，葡萄糖於發酵第6天後開始緩慢上升，果糖則於發酵液中緩慢累積。有機酸含量皆隨發酵時間持續累積，由於有機酸的累積，pH值從4.2下降至2.7直到發酵12天結束，因此建議紅茶菇發酵天數不宜太長。綜合上述，以5%含糖紅茶菇中添加0.1 %兒茶素於30 ℃下靜置發酵6天，可得最佳機能性之紅茶菇。(三) 本研究依不同比例混合小麥草汁及含糖紅茶湯液為發酵基質，接種菌酛後，接種量為20 % (v/v)，次於29±1 ℃下靜置培養12天。探討小麥草汁與含糖紅茶湯液混合比例對發酵液中酵母菌及醋酸菌菌數、總酚、總類黃酮，總花青素、各項酚類、糖類、有機酸、水溶性維生素含量及抗氧化特性之影響。結果顯示，各組酵母菌及醋酸菌分別於發酵至第6天及第3天達最大菌量，分別為7.2 log (cfu/mL)及5.1 log (cfu/mL)。混合小麥草汁之發酵液比傳統紅茶菇含有較高之總酚及類黃酮含量，並含有更多種類酚類成分，例如: 沒食子酸，兒茶素，咖啡酸，阿魏酸，槲皮素，綠原酸; 其有機酸(醋酸、葡萄糖酸及酒石酸)及水溶性維生素(B1、B2及泛酸)含量皆較未發酵前提升，維生素B1、B2及泛酸於發酵至第12天其等含量分別約為31.15~76.45 mg/100mL、44.71~76.10 mg/100mL及5.97~15.88 mg/100mL。在DPPH自由基清除能力可達90%以上; 氧自由基吸收能力 (oxygen radical absorbance capacity, ORAC)則由原先5.0 μmol trolox equivalent/mL提升至 12.8 μmol trolox equivalent/mL。以混合菌酛發酵後之含小麥草紅茶以培養基質比例為小麥草汁:紅茶=1:1 (v/v)之混合比例(初始蔗糖含量約4%, w/v)，經發酵6天所得之小麥草汁紅茶較未發酵前不僅顯著提升代謝產物如有機酸及水溶性維生素等含量，除獲得較多的營養成分，更有助於提升發酵液抗氧化力。經小麥草汁強化之紅茶經發酵後及抗氧化力。
本研究解明添加物之添加或基質的調整共同發酵而得的改良式紅茶菇發酵比傳統的及機能性成分(有機酸及酚酸)種類及含量，可以作為未來開發機能性飲品之參考。

Parallel Abstract 〈TOP〉

Kombucha is traditionally made by fermenting sugared black tea using a symbiotic culture of acetic acid bacteria and yeast. It contains many compounds with antioxidant activity, such as phenolics, water-soluble vitamins, organic acids, and minerals. Few studies have used materials besides tea leaves as the fermentation substrate for kombucha. The aim of this study is to create a novel kombucha and enhance cellulosic pellicle, phenolic compounds and antioxidant activities implementing herbal material or additive as the fermentation substrate.
Results showed that (1) The effects of adding caffeine (0%~0.4%, w/v) on the changes in viable cells, cellulose, antioxidant activity, and concentrations of both sugars and organic acids during fermentation were investigated. The results showed that the pH values decreased gradually from 3.9 to 2.8 with time. Viable counts of yeast and acetic acid bacteria reached their maximum values 7.2 log cfu/mL at Day 6, and 7.3 log cfu/mL at Day 3, respectively. The biosynthetic rates for the cellulosic pellicle were greater in the presence of caffeine, and the yields increased by up to 26.5% (based on wet weight). Adding caffeine significantly enhanced the production of gluconic acid (15.4g/L). Compared to traditional kombucha , adding catechin significantly also enhanced the antioxidant activity. These findings might be useful for developing noval beverages with beneficial effects. (2) The effects of adding (+)-catechin (0.1%~0.3%, w/v) on the changes in viable cells, cellulose, antioxidant activity, and concentrations of both sugars and organic acids during fermentation were investigated. The viable counts for acetic acid bacteria and yeast in the kombucha with added catechin were not significantly different, but their metabolic activities, particularly the production of gluconic acid, cellulose, and antioxidants, were enhanced. The biosynthetic rates for the cellulosic pellicle were greater in the presence of (+)-catechin, and the yields increased by up to 21.9% (based on wet weight). Compared to traditional kombucha (19.0 mg/mL), adding (+)-catechin significantly enhanced the production of gluconic acid , which ranged from 22.4 to 30.3 mg/ml. These findings might be useful for developing new beverages with beneficial effects. (3) Sweetened black tea (10% w/v) and wheatgrass juice (WGJ) were mixed in various ratios and used as fermentation substrate for enhancing phenolic compounds and antioxidant activity. Starter comprising of yeast (Dekkera bruxellensis), acetic acid bacteria (Gluconacetobacter rhaeticus and Gluconobacter roseus) was inoculated at 20% (v/v), and fermented statically at 29±1 ℃ for 12 days. The results viable counts of yeast and acetic acid bacteria reached their maximum values 7.2 log cfu/mL at Day 6, and 5.1 log cfu/mL at Day 3, respectively. All fermented products were characterized as having increased amounts of organic acids (acetic acid, gluconic acid, and tartaric acid), water- soluble vitamins (B1, B2, and pantothenic acid), total phenolic and flavonoid contents, but decrease in sugar contents after 12 days of fermentation. Contents of vitamins B1, B2, and pantothenic acid are 31.15-76.45 mg/100 mL, 44.71-76.10 mg/100 mL, and 5.97-15.88 mg/100mL, respectively. All WGJ-blended kombucha were characterized as having various phenolic compounds such as gallic acid, catechin, caffeic acid, ferulic acid, rutin, and chlorogenic acid as compared to traditional ones. Antioxidant activity of the modified kombucha was higher than that of traditional preparations. Addition of WGJ ensured the 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging ability of kombucha was greater than 90%, while the oxygen radical absorbance capacity increased from 5.0 to 12.8 μmol trolox equivalent/mL as the ratio of WGJ increased from 0 to 67% (v/v). The highest antioxidant activity was obtained using a 1:1 (v/v) black tea decoction to WGJ ratio and three days of fermentation, producing various types of phenolic acids. Our results suggest that intake of fermented black tea enhanced with wheatgrass juice is advantageous over unfermented one in terms of its nutritive values and might have more potential to reduce oxidative stress. 
In this study, it is possible to obtain the desired quality of beverage by blending traditional kombucha with herbal material or additive. This strategy ensures fermentation at faster rates without sacrificing quality. These findings might be useful for developing new beverages with beneficial effects.

Reference ( 129 ) 〈TOP〉

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