臺灣博碩士論文加值系統

本研究以台灣區收集之克弗爾菌元(kefir starter)為對象，除鑑定菌相組成及分析特性外，並嘗試以豆乳(soymilk)為原料，製作豆乳克弗爾，以期開發兼具二者優點之新產品。
自台灣收集之克弗爾，分離鑑定之乳酸菌為Lactobacillus helveticus與Leuconostoc mesenteroides，酵母菌為Kluyveromyces marxianus與Pichia fermentans。比較以上各菌株於牛乳中的生長特性，結果以Lb. helveticus之生長性最佳，而Kluy. marxianus之L(+)乳酸與乙醇產量較多。Leu. mesenteroides所產生的乳酸以D(-)型為主。P. fermentans雖不具有發酵乳糖的能力，但其蛋白分解活性極高，故於牛乳中仍可大量增殖。
在不同保存法對克弗爾菌元活性之影響方面，將克弗爾菌元以凍乾或未經凍乾的方式，分別存放於25、4及－20℃，再測定保存期間各菌元的發酵活性。結果顯示，克弗爾菌元中的乳酸菌於凍乾後尚可維持極高的活力，於－20℃保存9週再經發酵後，乳酸菌數與對照組間無顯著差異。而未經凍乾者，乳酸菌數則有隨貯藏時間延長而減少的趨勢。酵母菌對凍乾處理又較乳酸菌敏感。凍乾菌元製作之克弗爾，酵母菌數較未經凍乾者約減少90 %；而未凍乾即貯存於25℃及－20℃下，酵母菌均有明顯減少的趨勢，3週後已無酵母菌存在。各處理組之菌元產酸能力均有降低的現象，其中又以未凍乾而貯藏於25℃者最明顯。此外，凍乾對酒精產量亦有顯著降低的影響。故於9週貯藏期間內，以凍乾而凍藏於－20℃的方式，對克弗爾菌元的影響最小。
將克弗爾菌元接種於豆乳中，並探討添加不同糖類對克弗爾菌元生長性之影響。發現克弗爾於豆乳中，初始時乳酸菌數較接種於牛乳者高，但酵母菌數則較低。培養32小時後，豆乳中的酵母菌數、乳酸含量、酒精濃度均顯著地較牛乳者為低。添加1 %葡萄糖對於克弗爾發酵豆乳的乳酸菌數、酵母菌數、乳酸產量、酒精濃度，以及β半乳糖活性均有促進的效果，但對α半乳糖則有抑制的作用。克弗爾菌元於豆乳中生長時，其多醣產量較低。多醣之分子量為1.7 × 106，主要由葡萄糖與半乳糖組成，其中又以前者佔多數。豆乳經克弗爾菌元發酵後，主要風味成分以乙醛、丙酮、聯乙醯及乙醇為主，其含量均隨發酵時間而增加；而己醛含量則逐較減少。與發酵牛乳比較時，豆乳克弗爾僅有乙醛含量較高，其他風味成分則均較低。
克弗爾之機能性方面，探討牛乳及豆乳分別經克弗爾菌元發酵後，對小鼠體內腫瘤細胞之抑制性，以及對腸道生理機能的影響。結果顯示，餵食牛乳克弗爾或豆乳克弗爾均可減緩小鼠皮下腫瘤細胞生長，生長抑制率分別達64.8及70.9 %，且可觀察到腫瘤細胞有細胞凋亡現象，而小腸集合淋巴結亦受刺激活化。此外，腸道微生物中，雙叉乳酸桿菌(Bifidobacterium)均有顯著提高的現象，而糞便與小腸中總免疫球蛋白A (IgA)的含量亦增加。

This study was conducted to investigate the characterization and functional activity of milk and soymilk kefir.
The lactic acid bacteria isolated from kefir grains were identified as Lactobacillus helveticus and Leuconostoc mesenteroides, and the yeasts were identified as Kluyveromyces marxianus and Pichia fermentans. From the results of studying on the growth characteristics of the above strains, Lb. helveticus possessed better growth characteristics, and K. marxianus exhibited better L-lactic acid and ethanol production. D-lactic acid was the major form produced by Ln. mesenteroides. P. fermentans was a lactose-nonfermenting yeast, therefore the active proteolytic enzymes was essential for its growth in milk.
The lyophilized or non-lyophilized kefir grains were stored at 25, 4, and －20℃ for evaluated metabolic activity, and the results indicated that lyophilized grains maintained high level of lactic acid bacterial viabilities, and fermented milk obtained from these grains showed the same bacterial counts as fermented milk obtained from nonstored grains. Viabilities of lactic acid bacteria for non-lyophilized grains decreased with time. Yeast was more sensitive to lyophilization. Counts of yeast from lyophilized grains were about 90 % lower than those from nonstored grains. Counts of yeast from non-lyophilized grains stored at 25℃ or －20℃ decreased significantly, and yeast did not survive any more after 3 weeks storage. Acid production was low for each treatment except control. Lyophilization effected ethanol production by grains significantly. Storage at －20℃ is a better method to preserve lyophilized kefir grains.
For the manufacture of a new fermented product, the effects of added glucose, lactose, and sucrose on microbial growth, acid and ethanol production, and galactosidase activity in soymilk fermented with kefir grains were studied. Immediately after the addition of kefir grains to soymilk, the lactic acid bacterial counts were higher but the yeast counts were lower than in milk kefir. After fermentation for 32 hr, the concentrations of yeast, lactic acid, and ethanol in soymilk were significantly lower than those in milk kefir. Addition of 1% glucose to soymilk stimulated growth of lactic acid bacteria and yeast, the production of lactic acid and ethanol, and the β-galactosidase activity. Nevertheless α-galactosidase activity was suppressed by 1% glucose.
Soymilk kefir grains had lower polysaccharide contents than milk kefir grains, and its polysaccharide contained primarily glucose and galactose with the former predominating. The apparent molecular weight of the polysaccharide was estimated to be 1.7 ´ 106. In comparison to nonfermented soymilk, the concentrations of key volatile compounds, i.e., acetaldehyde, acetone, diacetyl, and ethanol increased, while n-hexanal decreased in soymilk kefir. The acetaldehyde level in soymilk kefir was slightly higher than for milk kefir, but the other volatile compounds were lower than for milk kefir.
The antitumor activities of milk kefir and soymilk kefir were studied in the mice subcutaneously inoculated with sarcoma 180 (S180). The growth of S180 solid tumor was inhibited, and the rates of inhibition were 65 % and 71 %, respectively. Peyer’s patches from milk kefir and soymilk kefir fed mice were activated. Furthermore, the counts of Bifidobacterium in cecum and feces, and the levels of total IgA in tissue extracts of the small intestinal wall as well as in fecal extracts were significantly higher in the milk and soymilk kefir groups than that of control group.

封面
壹、中文摘要
貳、緒言
參、文獻檢討
一、乳酸菌胞外多醣之組成與功能
二、發酵乳之抗癌性
三、黃豆之抗癌性
四、黃豆與血膽固醇
肆、實驗
第一章 克弗爾菌元中乳酸菌與酵母菌之分離鑑定
一、克弗爾菌元中微生物之菌相分佈
二、克弗爾菌元中乳酸菌之分離與鑑定
三、酵母菌之分離與鑑定
四、分離菌株之生長特性
第二章 克弗爾菌元之保存性
一、不同保存方式對菌元活力之影響
二、不同保存方式對菌元活性之影響
第三章 克弗爾菌元於豆乳中之生長性
一、乳酸菌於牛乳及豆乳中之生長性
二、酵母菌於牛乳及豆乳中之生長性
三、克弗爾菌元之乳酸生成量
四、克弗爾菌元之乙醇產量
五、克弗爾菌元之半乳糖活性
六、牛乳與豆乳克弗爾之黏度
第四章 牛乳與豆乳克弗爾之風味及多醣組成
一、牛乳與豆乳克弗爾粒之外觀型態
二、牛乳與豆乳克弗爾粒之微細構造
三、牛乳與豆乳克弗爾粒之化學組成
四、克弗爾粒之多醣組成及其分子量
五、克弗爾之揮發性物質組成
第五章 克弗爾之機能活性
第一節 克弗爾對小鼠腫瘤細胞生長之抑制
一、接種S180腫瘤細胞對小鼠生長之影響
二、S180細胞對臟器重量之影響
三、克弗爾對腫瘤組織生長之抑制
四、克弗爾對血液生化值之改善
五、克弗爾對腫瘤組織之影響
六、S180細胞對小鼠脾臟之影響
七、克弗爾對集合淋巴結之影響
第二節 克弗爾對小鼠腸道生理機能之影響
一、克弗爾對小鼠生長及臟器重量比之影響
二、克弗爾對小鼠腸內菌叢之影響
三、克弗爾對腸道IgA分泌量之影響
伍、結論
陸、參考文獻
柒、英文摘要
捌、作者小傳
玖、附錄

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