Study on production of sparkling lychee wine

荔枝為我國重要經濟果樹之ㄧ，由於產期短且不易保鮮，常造成產銷問題，故研製新產品以增加荔枝的加工利用有其必要。本研究以黑葉種荔枝為原料，探討荔枝氣泡酒之最適製程。 傳統氣泡酒製程為兩段式發酵，先經一次發酵(主發酵)製得基酒，再以二次發酵產生二氧化碳，使酒中富有細緻而持久之氣泡。主發酵條件經試驗後確認為荔枝原汁調糖至22 ºBrix，添加檸檬酸 (l.0 g/L) 及亞硫酸鹽(使游離二氧化硫濃度達200 ppm)，並接種Danstil D (Saccharomyces cerevisiae)，於25 ºC下發酵七天，製得荔枝發酵基酒。荔枝基酒再分別以傳統法，即添加25 g/L蔗糖及100 ppm亞硫酸鹽，並接種Fermivin PDM (Saccharomyces bayanus) 於15 ºC下進行二次發酵；或固定化酵母發酵法，即接種經3 %褐藻酸鈉固定化之酵母細胞進行二次發酵；或直接充填高壓二氧化碳使其最終壓力達5 kg/cm²，製成荔枝氣泡酒。 三種不同製程所得之成品經熟成六個月後分析比較，高壓氣體充填法所製得之荔枝氣泡酒氣泡粒徑較大；於濁度方面，以高壓氣體充填法及固定化酵母發酵法所製得之荔枝氣泡酒皆低於傳統發酵法；經喜好性品評試驗結果得知，在外觀方面，固定化酵母發酵法及高壓氣體充填法皆優於傳統法，在風味及口感方面，固定化酵母發酵法與傳統發酵法並無顯著差異，皆優於高壓氣體充填法。整體喜好性上，以固定化酵母發酵法有最高之接受度，其次為傳統法。確認二次發酵最適製程為荔枝基酒添加25 g/L蔗糖及100 ppm亞硫酸鹽，並接種經3 %褐藻酸鈉固定化之酵母Fermivin PDM ( Saccharomyces bayanus ) (膠體起始菌數：108 cells/mL，顆粒大小：2~3 mm) 8 g/每瓶600 mL於 15 ºC下進行發酵。以固定化酵母發酵法所製造之荔枝氣泡酒能免除傳統製程中的轉瓶步驟，縮短製程時間，並保有良好之風味口感。Lychee is an important fruit in Taiwan. There are some problems in production and marketing because lychee has a short harvesting period and difficult to store. So it is justified to improve its utilization by making new product. The project was to establish a suitable process for making sparkling lychee wine. Traditional method for the making of sparkling wine involves two steps of fermentation. The base wine is produced in the first step, called the primary fermentation. The secondary fermentation follows, and generates fine, sustainable bubbles in the wine. An adequate procedure for the primary fermentation was found to be adding sucrose, citric acid and sulfite to lychee juice to reach 22 ºBrix, 1.0 g/L and 200 ppm SO2 concentration, respectively, inoculating Saccharomyces cerevisiae Danstil D, and then incubating at 25 ºC for 7 days to obtain lychee base wine. The base wine can be added with 25 g/L sucrose and 100 ppm sulfite, then inoculated with Saccharomyces bayanus “Fermivin PDM” and held at 16 ºC for the traditional secondary fermentation, or inoculated with yeast cells bound in 3 % sodium alginate for the immobilized secondary fermentation. The base wine can also be infused with carbon dioxide to reach 5 kg/cm² pressure level in the production of sparkling wine in an alternative way. Both of the sparking wines produced by the carbon dioxide infusion method and by the immobilized yeast fermentation method have a lower turbidity than the one produced by the conventional fermentation method. There is no significant difference in flavor and mouth feel between the products from the immobilized yeast fermentation and from the conventional fermentation. However, both of the bubble stability and the acceptance of the sparkling wine produced by the carbon dioxide infusion method are inferior to those of the products produced by the other two methods. We concluded that to produce sparkling lychee wine by immobilized yeasts enjoys the advantage of being exempted from the tedious riddling operation in the conventional method, shortening the process time, yet retaining good flavor and taste; and therefore is worth to be recommended.目錄 中文摘要 .................................................I 英文摘要 ................................................II 壹、前言 .................................................1 貳、文獻整理..............................................2 一、氣泡酒................................................2 二、氣泡酒之類型..........................................3 三、固定化生物技術.......................................13 1.固定化方法之區分.....................................13 2.固定化酵母細胞於製酒上的應用.........................16 参、材料與方法...........................................18 一、實驗材料.............................................18 二、儀器設備.............................................19 三、實驗架構.............................................20 四、實驗方法.............................................23 1、荔枝果汁之製備......................................23 2、主發酵條件試驗......................................23 3、二次發酵條件試驗....................................25 (1)、傳統二次發酵法...................................25 (2)、固定化酵母發酵法.................................27 4、高壓氣體充填試驗....................................29 5、熟成試驗............................................29 五、分析方法.............................................30 1、可溶性固形物........................................31 2、酸鹼值..............................................31 3、可滴定酸............................................31 4、揮發性酸............................................32 5、總糖................................................32 6、總酚類化合物........................................34 7、酒精度..............................................34 8、色澤 ................................................34 9、濁度................................................35 10、感官品評............................................35 11、懸浮粒徑之測定......................................35 12、氣體壓力測定........................................35 13、有機酸分析..........................................36 14、香氣成分分析及鑑定..................................36 .肆、結果與討論..........................................40 一、主發酵...............................................40 1、荔枝果汁之成分分析..................................40 2、荔枝發酵基酒於主發酵期間之成分變化..................41 3、建立荔枝發酵基酒製程................................51 二、二次發酵.............................................61 三、氣泡粒徑分析.........................................72 四、香氣成分分析.........................................79 五、品評試驗.............................................84 伍、結論.................................................89 陸、參考文獻.............................................91 圖目錄 圖一、溫度及壓力對酒中二氧化碳含量之影響..................5 圖二、三種氣泡酒製程之流程圖..............................9 圖三、傳統法氣泡酒製程...................................10 圖四、酵母菌自解過程所產生之各種物質對氣泡酒品質之影響...11 圖五、酵素或微生物細胞的各種固定法.......................15 圖六、以傳統法及固定細胞進行氣泡酒瓶中發酵之流程圖.......17 圖七、荔枝發酵基酒之製程.................................24 圖八、傳統氣泡酒之發酵製程...............................26 圖九、酵母細胞固定化流程圖...............................28 圖十、高壓氣體充填機裝置圖...............................30 圖十一、揮發性酸測定之裝置圖.............................33 圖十二、氣體壓力測定.....................................37 圖十三、不同酵母對荔枝酒發酵基質於發酵過程中可溶性固形物 含量之影響.......................................44 圖十四、不同酵母對荔枝酒發酵基質於發酵過程中酒精含量之影響 ................................................45 圖十五、不同酵母對荔枝酒發酵基質於發酵過程中可滴定酸含量之 影響 ............................................46 圖十六、不同酵母對荔枝酒發酵基質於發酵過程中pH值之影響...47 圖十七、不同酵母對荔枝酒發酵基質於發酵過程中揮發性酸含量之 影響.............................................48 圖十八、檸檬酸添加量對荔枝發酵基酒於發酵過程中總糖含量之影 響...............................................53 圖十九、檸檬酸添加量對荔枝發酵基酒於發酵過程中酒精含量之影 響...............................................54 圖二十、檸檬酸添加量對荔枝發酵基酒於發酵過程中pH值之影響 .................................................55 圖二十一、檸檬酸添加量對荔枝發酵基酒於發酵過程中可滴定酸含 量之影響.........................................56 圖二十二、檸檬酸添加量對荔枝酒發酵基質於發酵過程中揮發性酸 含量之影響.......................................57 圖二十三、荔枝發酵基酒之製程.............................59 圖二十四、Pasture Champagne及Fermivin PDM對荔枝氣泡酒二次發 酵期間壓力之影響.................................62 圖二十五、Pasture Champagne及Fermivin PDM對荔枝氣泡酒二次發 酵期間揮發性酸含量之影響.........................63 圖二十六、Pasture Champagne及Fermivin PDM對荔枝氣泡酒二次發 酵期間A420之影響.................................64 圖二十七、Pasture Champagne及Fermivin PDM對荔枝氣泡酒二次發 酵期間A520之影響.................................65 圖二十八、二氧化硫添加量對荔枝氣泡酒於25 ºC熟成期間褐變程度 之影響...........................................68 圖二十九、二氧化硫添加量對荔枝氣泡酒於25 ºC熟成期間紅變程度 之影響...........................................69 圖三十、固定化於褐藻酸鈉膠體之不同酵母量對二次發酵期間氣體 壓力之影響.......................................70 圖三十一、荔枝基酒中懸浮物質粒徑分布圖...................74 圖三十二、荔枝基酒中懸浮物質重量分布圖...................75 圖三十三、傳統法發酵法之氣泡酒中懸浮粒子粒徑分布圖.......76 圖三十四、固定化發酵法之氣泡酒中懸浮粒子粒徑分布圖.......77 圖三十五、充填法之氣泡酒中懸浮粒子粒徑分布圖.............78 圖三十六、荔枝氣泡酒之最適製程...........................90 表目錄 表一、各國對氣泡酒壓力之規範..............................4 表二、氣泡酒依調糖程度分類...............................12 表三、歷年荔枝果汁物化特性分析之比較.....................43 表四、酒中揮發性酸標準...................................49 表五、荔枝基酒外觀性質分析...............................58 表六、荔枝基酒基本性質分析...............................60 表七、不同酵母固定化膠體添加量對二次發酵壓力與混濁度之影響 ...................................................71 表八、不同菌種發酵所得荔枝氣泡酒之香氣成份鑑定...........81 表九、不同熟成時間所得荔枝氣泡酒之香氣成份鑑定...........82 表十、不同製程所得荔枝氣泡酒之香氣成份鑑定...............83 表十一、荔枝氣泡酒基本成分分析...........................86 表十二、填充法之不同糖酸比嗜好性品評.....................87 表十三、不同製程所得荔枝氣泡酒之嗜好性感官品評...........8