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研究生:彭信蒼
研究生(外文):Hsin-Tsang Peng
論文名稱:利用納豆菌、酵母菌及米麴菌發酵糙米
論文名稱(外文):Use of Bacillus subtilis var. natto, Saccharomyces cerevisae, and Aspergillus oryzae in fermentation of brown rice
指導教授:王正仁王正仁引用關係
指導教授(外文):Chen-Jen Wang
學位類別:碩士
校院名稱:大葉大學
系所名稱:分子生物科技學系碩士班
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
論文頁數:99
中文關鍵詞: 米麴菌 酵母菌 心血管 消化性
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  • 被引用被引用:1
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糙米是營養豐富的食物,可以促進新陳代謝,預防動脈硬化、癌症、心血管疾病和貧血症,但糙米因被周邊纖維組織包埋,不易被人體消化吸收,且風味不佳;若經長時間蒸煮,則其營養素因過度加熱而損失。但若利用納豆菌(Bacillus subtilis var. natto)及米麴菌(Aspergillus oryzae)的多種水解酵素分泌能力及釀酒酵母菌(Saccharomyces cerevisiae)的風味產生能力,當能解決糙米的難消化問題,保存糙米的營養,提高消化性,並提升多樣的風味與保健成分。 因此,本研究的目的是以熟糙米為原料,利用多菌株發酵技術,生產機能性發酵糙米,並對發酵糙米進行機能性成分分析,以建立發酵糙米的生產依據。
菌種的製備方式及接種量對發酵影響的結果顯示,以酵母麥芽精培養液(yeast and malt extract broth,YMB)當作製備培養液且個別製備菌種時,再以10%的接種量接至糙米中發酵,可得較高納豆菌孢子數、酵母菌數及葡萄糖胺含量。糙米中的水添加量比例對糙米發酵的影響顯示,以1:0.9的比例進行發酵可以得到最多的納豆菌孢子數。而不管以單菌、雙菌或三菌發酵,納豆菌孢子數均可達108 CFU/g,酵母菌亦可達108 CFU/g,尤其當酵母菌與米麴菌一起發酵時,酵母菌數更可高達109 CFU/g。米麴菌只有與納豆菌同時發酵時,葡萄糖胺的含量較低,其餘皆可達3 mg/g。超氧歧化酶(superoxide dismutase)活性以酵母菌與米麴菌一起發酵時最高,有33 U/g。自由基清除率則以單一米麴菌或酵母菌與米麴菌及三菌發酵時最高,可達30%以上。糙米中蛋白酶活性以三菌同時發酵最高,達13.42 U/g。添加葡萄糖或酵母粉則對菌數或其他成分無顯著助益。

關鍵字:糙米、納豆菌、釀酒酵母菌、米麴菌、超氧歧化酶、自由基清除率、蛋白酶、葡萄糖胺。
Brown rice is a nutritious food, capable of enhancing metabolism and preventing arterial sclerosis, cancer, cardiovascular's disease and anaemia. However, its nutritious ingredients are mostly bound by peripheral tissues and thus not be easily digested or have a lower bioavailability. If it is overly cooked, its nutritious components may disapeear due to overheating. But if use of the ability of secreting various hydrolytic enzymes by Bacillus nato (BN) and Aspergillus oryzae (AO) with that of forming varied flavors by Saccharomyces cerevisiae (SC), the indigestibility of brown rice may be solved and its nutrition may well be preserved. In the mean time, its digestibility will be significantly strengthened and various flavors and healthy components are additionally formed in it. Therefore, the purpose of this study is to develop a fermentation process for production of functional fermented food from brown rice by mixed-culture fermentation. The functional components of fermented brown rice and its physiological function will be investigated too. These results can provide information for establishment of a fermentation process for production of functional fermented brown rice.
Study on the effect of starter preparation method and inoculum size on brown rice fermentation shows that use of YMB (yeast and malt extract broth) as starter cultivation broth and 10% of inoculum size resulted in the highest BN spore and SC cell numbers with a substantial amount of glucosamine in fermented brown rice. A 1:0.9 of brown rice to water ratio led to the best formation of BN spores. No matter what single culture, double culture or mixed culture was used in fermentations, the final numbers of BN spores and SC cells could all reach 108 CFU/g. When both SC and AO were simultaneously used in fermentations, the final number of SC cells even reach 109 CFU/g. Only when both BN and AO were simultaneously used in fermentations, glucosamine content in harvested brown rice reached the least. And for other cases, glucosamine content in harvested brown rice could all reach 3mg/g. The SOD (superoxide dismutase) activity reached the highest value of 33U/g when both AO and SC were used in fermentation. The DPPH free radical scavenging ability could exceed 30% when single AO or both AO and SC or triple culture was inoculated. When three starters were simultaneously used, the highest protease activity could be obtained with 13.42U/g. Supplementation of glucose or yeast powder in brown rice could not help production of functional ingredients.

Key words: brown rice, Bacillus natto, Saccharomyces cerevisae, Aspergillus oryzae, SOD, DPPH, protease, glucosamine.
封面內頁
簽名頁
授權書 iii
中文摘要 iv
英文摘要 vi
致謝 vii
目錄 x
圖目錄 xii
表目錄 xv

1. 緒言 1
2. 文獻回顧 3
2.1 稻米 3
2.1.1 稻米之分類 3
2.1.2 米的營養成份 5
2.2 糙米(brown rice) 5
2.2.1 糙米之功能性物質 6
2.2.2 米糠醇 7
2.2.3 植酸 7
2.2.4 肌醇 8
2.2.5 γ-胺基丁酸(γ-amino butyric acid, GABA) 9
2.3 混合菌發酵(mixed culture fermentation) 9
2.4 菌種介紹 10
2.4.1 麴菌屬(Aspergillus) 10
2.4.2 米麴菌(Aspergillus oryzae) 11
2.4.3 枯草芽孢桿菌(Bacillus subtilis) 12
2.4.4 納豆菌(Bacillus subtilis var. natto) 12
2.4.5 酵母(yeasts) 13
2.4.6 釀酒酵母菌(Saccharomyces cerevisae) 14
2.5 機能性成分 14
2.5.1 超氧歧化酶(superoxide dismutase) 14
2.5.2 1,1-diphenyl-2-picryl-hydrazil (DPPH) 15
2.5.3 蛋白酶(protease) 15
2.5.3.1 蛋白酶的應用 16
3. 材料與方法 17
3.1 實驗材料 17
3.1.1 糙米 17
3.1.2 發酵菌種 17
3.2 儀器設備 17
3.3 培養基與藥品 18
3.3.1 培養基 18
3.3.2 藥品 20
3.4 藥品配製 21
3.4.1 HPLC系統用移動相 21
3.4.2 自由基清除率分析 22
3.5 實驗方法 22
3.5.1 發酵方法 22
3.5.2 發酵接種液製備 23
3.5.2.1 分別培養 23
3.5.2.2 混合培養 24
3.5.3 發酵菌接種量 24
3.5.3.1 單菌發酵 24
3.5.3.2 雙菌發酵 25
3.5.3.3 三菌發酵 25
3.5.4 糙米與水添加量的比例 25
3.6 分析方法 26
3.6.1 含水量測試 26
3.6.2 粗酵素萃取 26
3.6.3 納豆菌孢子數 26
3.6.4 酵母菌菌數 27
3.6.5 米麴菌生長測量 27
3.6.6 超氧歧化酶(SOD)活性 29
3.6.7 自由基清除率(DPPH) 29
3.6.8 蛋白酶活性 30
3.6.9 發酵糙米上頭之氧氣百分比 30
3.6.10感官分析 31
4. 結果與討論 32
4.1 菌種製備方式及接種量之探討 32
4.2 水添加量比例對混合菌發酵糙米之影響 33
4.3 其他發酵菌種對發酵糙米中納豆菌生長的影響 34
4.4 其他發酵菌種對發酵糙米中酵母菌生長的影響 35
4.5 其他發酵菌種對發酵糙米中米麴菌生長的影響 37
4.6 單菌或多菌株發酵對發酵糙米中超氧歧化酶活性的 39
4.7 單菌或多菌株發酵對發酵糙米中自由基清除活性的影
響 40
4.8 單菌或多菌株發酵對發酵糙米中蛋白酶的影響 41
4.9 葡萄糖及酵母粉對發酵表現的影響 42
4.10 感官分析 43
5. 結論 45
6. 參考文獻 78
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