臺灣博碩士論文加值系統

中文摘要
長期攝食的高膽固醇與高油脂飲食易引發高血脂症 (hyperlipidemia) 與肥
胖，進而導致粥狀動脈硬化、糖尿病與癌症等疾病。1979 年已有研究指出，紅
麴次級代謝物 monacolin K 具有降低膽固醇之效果，monascin 具有抗發炎、
ankaflavin 具有抗癌之功效。深層海水中具有豐富的鈣、鎂、磷酸根等離子，具
有促進酵母菌生長或麴菌的活性功能。紅麴與深層海水之過去研究均證實具有降
低體脂肪之功效。本研究以深層海水為水分來源發酵之紅麴山藥，進行降血脂與
降低體脂肪之保健功效評估，並以反應曲面法探討紅麴黃色素 monascin 與
ankaflavin 之固態發酵的最適培養條件。保健功效評估以高膽固醇飲食之倉鼠進
行降血脂之動物試驗及高油脂飲食之大鼠進行降低體脂肪試驗。在降血脂試驗的
研究結果顯示，餵食深層海水發酵之紅麴山藥一倍劑量可顯著降低高膽固醇飲食
組中的血清中總膽固醇、三酸甘油酯、低密度脂蛋白膽固與脂質過氧化程度，且
具有提高高密度脂蛋白膽固醇與預防動脈粥狀硬化之效果，且對肝臟不具毒性。
其中與以逆滲透水發酵之紅麴山藥比較，在降低脂質過氧化程度與心臟主動脈脂
質斑塊方面具有更顯著之效果。體脂肪方面，深層海水發酵之紅麴山藥與逆滲透
水發酵之紅麴山藥比較，在體重變化量、總體脂率、脂肪細胞截面積、血清中的
三酸甘油酯與肝臟中總膽固醇含量具有更顯著的降低效果。由降低血脂與體脂肪
試驗中發現，monascin 與 ankaflavin 均為降低血脂與體脂肪之功效成分，進而
以反應曲面探討其固態發酵的最適培養條件。研究結果發現，moanscin 與
ankaflavin 在 300 g 山藥基質進行發酵，於加水量 80 mL、pH 值為 3~4 之間
與深層海水含量越高的培養條件下，均具有較高的生成量。另一方面，紅麴菌所
產生的肝腎毒素— citrinin，其含量需越低越好，而在最適培養條件中發現，
citrinin 在加水量 80 mL、pH 值3~4 之間與深層海水含量較高的培養條件之
下，具有較低的生成量。因此，選取加水量 80 mL、pH 值為 3.2 與深層海水
含量為 89% 作為培養條件，結果可得可得 monascin 含量為 7933 mg/kg、
ankaflavin 為 2591 mg/kg 與 citrinin 含量為 1720 ppb。

Dietary containing high cholesterols and excessive fats for a long period are
easily contributable to hyperlipidemia and obesity, which lead to atherosclerosis,
diabetes, cancers and other diseases and disorders. Recent studies has indicated that
monacolin K, monascin and ankaflavin, the Monascus secondary metabolites,
expressed cholesterol-lowering effect, anti-inflammation and anti-cancer, respectively.
Calcium ions, magnesian ions, and phosphate ion as well as other ions contained in
deep seawater are able to stimulate the growth of yeast and Aspergillus. However, the
previous researches have proved that both Monascus and deep sea water reduced body
fats. In this study, deep sea water was used as the water source for the fermentation of
red mold dioscorea (known as the Monascus fermented dioscorea). We further carried
out assessment on hypolipidemic effect and anti-obesity effect. After confirming the
functional ingredients, the optimal culture conditions of solid fermentation for the
production of functional ingredients were investigation by using response surface
methodology (RSM). In the assessment on healthcare effects, hypolipidemic hamsters
induced by fed high cholesterol diet and obesity rat induced by fed high fat diet were
used as animal model of hyperlipidemic and anti-obesity test, respectively. The results
of hyperlipidemic experiments showed that one-fold dose of RMD fermented using
deep sea water (DSW-RMD) would perform significant decrease in the levels of total
cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and
lipids, raising in the levels of high-density lipoprotein cholesterol (HDL-C),
atherosclerosis prevention, as well as without damage for liver tissue. The results of
body fat-lowering experiments revealed that DSW-RMD performed more significant
lowering effect on weight gain, total fat ratio, the crossing area of adipose cells, serum
TG, and liver TC than RO-RMD. In addition, both monascin and ankaflavin should be
the functional ingredients of hypolipidemic and body fat-lowering effect. Therefore,
the optimal culture condition for higher monascin and ankaflavin production and
lower citrinin concentration was investigated further. High moanscin and ankaflavin
production and lower citrinin concentration was observed as using the condition with
80mL water added of deep sea water at pH 3.2. The RSM condition (80 mL water
added, 89% DSW, pH 3.2) resulted in an optimal result with higher monascin and
ankaflavin at least 7933 mg/kg and 2591 mg/kg, as well as lower citrinin below 1720
ppb.

目錄
中文摘要……………………………………………………………………………..Ⅰ
英文文要……………………………………………………………………………..Ⅱ
縮寫表………………………………………………………………………………..Ⅲ
前言……………………………………………………………………………………1
第一章 文獻回顧……………………………………………………………………4
第一節 深層海水之介紹與保健功效……………………………………………..4
第二節 紅麴菌之介紹與保健功效………………………………………………..9
第二章 實驗動機與實驗架構……………………………………………………..20
第三章 材料與方法………………………………………………………………..22
第一節 實驗材料……………………………………………………………...….22
第二節 藥劑………………………………………………………………………23
第三節 紅麴山藥之培養條件……………………………………………………24
第四節 Monascin、ankaflavin 與 citrinin 之分析方法……………………….26
第五節 以反應曲面法探討最適培養條件之實驗設計…………………………27
第六節 體內動物試驗……………………………………………………………30
第四章 研究結果……………………………………………………………………42
第一節 深層海水之成分分析……………………………………………………42
第二節 不同水分來源發酵之紅麴山藥次級代謝物分析………………………42
第三節 降低體內血脂肪之動物實驗評估………………………………………42
第四節 降低體脂肪之動物實驗評估……………………………………………59
第五節 以 RSM 探討 monascin 與 ankaflavin 之最適培養條件…………...74
第五章 討論…………………………………………………………………………83
第六章 結論…………………………………………………………………………86
第七章 未來研究方向………………………………………………………………88
第八章 參考文獻........................................................................................................89
附錄、已發表之研討會論文…………………………………………………………94
表目錄
表 1-1 深層海水之營養元素及其功能………………………………………….....7
表 1-2 深層海水之功能與應用產品…………………………………………...…10
表 3-1 三變數-三階層反應曲面設計之操作條件變數及階層………………….28
表3-2 固態發酵之三因子-三階層之中心旋轉組合設計………………………..29
表3-3 實驗動物 Syrian 倉鼠分組與樣品劑量之配置………………………….32
表 3-4 降血脂動物試驗各組之飼料配方組成…………………………………...33
表3-5 實驗動物 SD 大鼠分組與樣品餵食劑量之配置………………………..38
表3-6 降低體脂肪動物試驗各組之飼料配方組成………………………………39
表4-1 深層海水之成分分析………………………………………….…………...43
表4-2 紅麴次級代謝物成分分析…………………………………………………49
表4-3 餵食不同試驗物質對倉鼠血清中 TC、TG、HDL-C 與 LDL-C 含量之
影響…………………………………………………………………………………..51
表 4-4 不同試驗物質對倉鼠血清中 AST 與 ALT 活性之影響……………...53
表 4-5 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠體重之影響……….60
表 4-6 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠的總熱量攝取、攝食
量及利用率之影響…………………………………………………………………..61
表 4-7 不同試驗物質對高油脂飲食之雄性 SD 大鼠總體脂肪重量與體脂率之
影響…………………………………………………………………………………..63
表 4-8 不同試驗物質對高油脂飲食之雄性 SD 大鼠腎周圍體脂肪與體脂率之
影響…………………………………………………………………………………..64
表 4-9 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠副睪周圍體脂肪與體
脂率之影響…………………………………………………………………………..65
表 4-10 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠脂肪細胞截面積之
影響…………………………………………………………………………………..67
表 4-11 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠血清脂質之影響…68
表 4-12 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠血糖含量之影響...69
表 4-13 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠肝、腎指數之影響.71
表 4-14 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠電解質平衡之影
響……………………………………………………………………………………..72
表 4-15 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠酮體濃度與肌酸酶
之影響………………………………………………………………………………..73
表 4-16 餵食不同試驗物質對高油脂飲食之雄性 SD 大鼠肝臟脂質含量之影
響……………………………………………………………………………………..75
表 4-17 單因子培養條件對紅麴次級代謝物之影響……………………………76
表 4-18 RSM 驗證結果…………………………………………………………..82
表 4-19 實驗結果綜合比較…………………………………………………….…87
圖目錄
圖1-1 深層海水的生成過程圖…………………………………………………….5
圖1-2 紅麴菌之生活史…………………………………………………………...12.
圖 1-3 八種紅麴色素之化學結構………………………………………………..14
圖 1-4 Dimerumic acid 抗氧化機制……………………………………………..17
圖 2-1 實驗大綱架構圖…………………………………………………………..21
圖 4-1 紅麴次級代謝物 monascin 與 ankaflavin 之紫外光全波長吸收波峰圖
譜……………………………………………………………………………………44
圖 4-2 紅麴次級代謝物 monascin、ankaflavin 與 citrinin 之 HPLC 層析
圖……………………………………………………………………………………45
圖 4-3 Monascin 之 HPLC 檢量線…………………………………………….46
圖 4-4 Ankaflavin 之 HPLC 檢量線…………………………………………...47
圖 4-5 Citrinin 之 HPLC 檢量線……………………………………………….48
圖 4-6 餵食不同試驗物質對倉鼠血清中脂質過氧化程度之影響……………..55
圖4-7 餵食不同試驗物質之高膽固醇倉鼠心臟主動脈脂質斑塊………………57
圖4-8 餵食不同試驗物質對於倉鼠胸主動脈中脂質堆積程度之影響………58
圖 4-9 培養 pH 與深層海水比例對紅麴次級代謝物 monascin 之反應曲面
圖……………………………………………………………………………………..77
圖 4-10 培養 pH 值與深層海水比例對紅麴次級代謝物 ankaflavin 之反應曲面
圖……………………………………………………………………………………..78
圖 4-11 培養 pH 與深層海水比例對紅麴次級代謝物 citrinin 之反應曲面
圖……………………………………………………………………………………..80
圖 4-12 培養 pH 與深層海水比例對紅麴次級代謝物 monascin、ankaflavin 與
citrinin 生成量之等高線交疊圖……………………………………………………81

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