臺灣博碩士論文加值系統

部份由植物中分離出的多醣由於具有促進乳酸菌生長的能力，因此被認為是一種益菌生物質(prebiotic)。而菇類多醣長久以來即因為免疫調節、抗腫瘤以及抗氧化等功效而廣為人知，但目前對於菇類多醣是否可做為益菌生物質了解仍不多。本研究利用香菇柄、杏鮑菇基部以及金針菇基部等做為多醣萃取來源，探討菇類多醣是否能提升乳酸益生菌(probiotic) Lactobacillus acidophilus BCRC 10695、Lactobacillus casei BCRC14080以及Bifidobacterium longum BCRC 14664於低溫儲存下之存活率，以及在腸胃道試驗中提升其對胃酸及膽鹽之抵抗能力，並評估添加多醣之發酵乳中的益生菌存活率以及是否有助增加發酵乳機能性。

實驗結果顯示，添加菇類多醣可以減緩三株測試乳酸菌在於低溫儲存下存活菌數減少之速率，中低劑量之多醣添加量(0.1%、0.5%(w/w))較添加高劑量(1.0%)者更能達到延緩乳酸菌死滅的效果。而添加0.5%多醣也可以改善乳酸菌對胃酸及膽鹽的抵抗能力，在同樣測試時間下，添加多醣之試驗組其存活菌數高於未添加多醣之控制組。

於低溫儲存試驗中，菇類多醣對發酵乳中益生菌之助益因菌種而異，其可維持L. acidophilus BCRC 10695於低溫下有較高存活菌數、可提升L. casei BCRC 14080之存活菌數以及減緩B. longum BCRC 14664存活菌數之減少速率，研究結果證明菇類多醣可幫助發酵乳中的乳酸菌，於低溫儲存下維持其存活率。

而在發酵乳蛋白質水解活性方面，含有B. longum BCRC 14664的發酵乳在添加金針菇多醣(FVP, Flammulina velutipes polysaccharide)後，於儲存期第28天時達到本次實驗中最高的蛋白質水解；ACE（Angiotensin I converting enzyme）抑制活性方面，發酵乳中的ACE抑制活性最高約為20%，且ACE抑制活性的變化與添加的多醣種類或儲存的天數沒有明顯的相關性，

本研究證實了藉由添加菇類多醣，可以提升益生菌在低溫儲存下的存活率、改善益生菌對胃酸及膽鹽的抵抗能力以及提升發酵乳中的益生菌存活率以及增進發酵乳的機能性，菇類多醣確實具有當做益菌生物值的潛力。

Some plant polysaccharides had been considered as having prebiotic effect which can selectively stimulate the growth of probiotics. The mushroom polysaccharides are well known for their immunomodulatory, anti-tumor and anti-cancer activity, however, litter information is available showing if they could be used as prebiotics. In this study, stipes of Lentinula edodes, bases of Pleurotus eryngii and Flammulina velutipes were used as raw materials for polysaccharides extraction. The effect of mushroom polysaccharides on viability of Lactobacillus acidophilus BCRC 10695, Lactobacillus casei BCRC14080 and Bifidobacterium longum BCRC 14664 under refrigerated storage and on their gastrointestinal tolerance were investigated. The viability of probiotics, proteolysis and angiotensin-converting enzyme (ACE) inhibitory activity of the fermented milk, which supplemented with mushroom polysaccharides were also investigated during cold storage.

Results showed that mushroom polysaccharides enhanced survival of probiotics in MRS broth under refrigerated storage. Supplemented mushroom polysaccharides in lower concentrations (0.1-0.5% (w/w)) had better effect to enhance the probiotics viability than in higher concentration (1.0% (w/w)). The survival rate of probiotic bacteria in simulated gastric acid (pH 2.0) and bile juice (4.5%. pH 8.0) was higher compared to control group. Probiotic bacteria with 0.5% (w/w) mushroom polysaccharides supplementation improved their tolerance to such conditions.

Supplementation with mushroom polysaccharides could keep the higher microbial counts of L. acidophilus BCRC 10695, improve viability of L. casei BCRC 14080 and retard the death of B. longum BCRC 14664 in fermented milk during cold storage.

The presence of high functionality of proteolysis in fermented milk containing B. longum with 0.1 % (w/w) FVP (F. velutipes polysaccharide) at 28 days in cold storage was observed; 20% ACE inhibition was observed in the fermented milk In addition, the presence of prebiotics or storage period did not influence the ACE inhibitory activity.

This study indicated that mushroom polysaccharides enhanced probiotics survival in MRS during cold storage and improve their tolerance in simulated gastric and bile juice. Supplementation with mushroom polysaccharides also increased the population of probiotics and functionality of the fermented milk, which suggest that mushroom polysaccharides are potential prebiotics.

目錄 v
圖次目錄 viii
表次目錄 x
縮寫對照表 xi
壹、前言 1
貳、文獻回顧 3
一、常見食藥用菇類之成份、生理活性及市場概況介紹 3
(一)、菇類介紹 3
(二)、菇類產品市場現況 3
(三)、臺灣市場上常見之菇類 4
(四)、菇類的生理活性 10
(五)、菇類多醣的功能特性 14
(六)、菇類下腳 20
二、乳酸菌之介紹 22
(一)、乳酸菌之定義 22
(二)、乳酸菌之分類 23
(三)、生理機能活性 26
(四)、ABC三益菌 28
三、發酵乳製品 30
(一)、定義及種類 30
(二)、相關機能性 31
(三)、發酵乳之品質維持 34
四、Probiotics、Prebiotics、Synbiotics介紹 35
(一)、Probiotics 35
(二)、Prebiotics 36
(三)、Synbiotics 38
參、實驗架構 42
肆、材料與方法 43
一、實驗材料 43
(一)、試驗用菌種 43
(二)、菇柄樣品 43
(三)、藥品與培養基 43
(四)、培養基與緩衝溶液配置 46
(五)、儀器設備 49
(六)、電腦套裝軟體 50
二、實驗方法 51
(一)、菌種活化及街種源之製備 51
(二)、菇柄粉末及水萃多醣之製備 51
(三)、MRS培養低溫儲存試驗 52
(四)、耐酸耐膽鹽試驗 53
(五)、乳酸菌株選擇性培養 54
(六)、發酵乳低溫儲存試驗 55
(七)、發酵乳機能性探討 57
(八)、統計分析 59
三、分析方法 60
(一)、粗萃多醣總醣量測定 60
(二)、乳酸菌數計算 60
(三)、發酵乳pH測定 61
(四)、ACEI活性計算 61
伍、結果與討論 66
一、菇柄水萃多醣之產率 66
二、粗萃多醣總醣含量 67
三、低溫冷藏試驗 67
(一)、不同濃度多醣添加量探討 68
(二)、不同種類多醣添加試驗 70
(三)、小結 72
四、耐酸耐膽鹽試驗 74
五、選擇性培養基測試 78
六、乳低溫儲存試驗 79
七、發酵乳機能特性探討 84
陸、結論 107
柒、未來展望 109
捌、參考文獻 110



圖次目錄
圖一、具抗腫瘤活性之含β-(1→6)支鏈的β-(1→3)主鏈-D葡聚醣結構 18
圖二、由β-葡聚醣投藥產生的寄主免疫反應 19
圖三、香菇、杏鮑菇及金針菇之柄部或基部 21
圖四、各種微生物之葡萄糖代謝途徑 24
圖五、益生菌的健康功效 39
圖六、酚-硫酸法之葡萄糖檢量線 63
圖七、(A) Spiral plater計數盤；(B) 利用spiral plater所製作之plate 64
圖八、Hippuric acid之HPLC檢量線 65
圖九、乳酸益生菌培養於添加不同濃度香菇多醣的MRS培養基於低溫儲存下28
天的存活菌數變化 89
圖十、乳酸益生菌培養於添加0.1%菇類多醣的MRS培養基於低溫下儲存28天
的存活菌數變化 90
圖十一、乳酸益生菌培養於於添加0.5%菇類多醣的MRS培養基於低溫下儲存
28天的存活菌數變化 91
圖十二、經過菇類纖維固定化的L. acidophilus BCRC 10695培養於添加0.5%菇
類多醣的胃酸模擬液中180分鐘的存活菌數 92
圖十三、經過菇類纖維固定化的L. casei BCRC 14080培養於添加0.5%菇類多醣
的胃酸模擬液中180分鐘的存活菌數 93
圖十四、經過菇類纖維固定化的B. longum BCRC 10695培養於添加0.5%菇類多
醣的胃酸模擬液中180分鐘的存活菌數 94
圖十五、經過菇類纖維固定化的L. acidophilus BCRC 10695培養於添加0.5%菇
類多醣的膽鹽模擬液中0分鐘及240分鐘的存活菌數 95
圖十六、經過菇類纖維固定化的L. casei BCRC 14080培養於添加0.5%菇類多醣
的膽鹽模擬液中0分鐘及240分鐘的存活菌數 96
圖十七、經過菇類纖維固定化的B. longum BCRC 10695培養於添加0.5%菇類多
醣的膽鹽模擬液中0分鐘及240分鐘的存活菌數 97
圖十八、含L. acidophilus BCRC 10695之發酵乳於儲存期間蛋白質水解活性變化 102
圖十九、含L. casei BCRC 14080之發酵乳於儲存期間其蛋白質水解活性變化 103
圖二十、含B. logum BCRC 14664之發酵乳於儲存期間其蛋白質水解活性變化 104
圖二十一、含L. casei BCRC 14080之發酵乳於儲存期間其ACE抑制活性變化 105
圖二十二、含B. longum BCRC 14664之發酵乳於儲存期間其ACE抑制活性變化 106



表次目錄
表一、台灣各種菇類生產概況 7
表二、台灣菇類在批發市場交易量 8
表三、香菇在醫療上之應用 9
表四、菇類與其他食品營養價值比較 12
表五、菇類的藥效及機能性成分 13
表六、自食藥用菇類中所分離得具抗腫瘤活性多糖 17
表七、同型發酵和異型發酵之乳酸菌區分 25
表八、生產傳統發酵乳製品所採用的微生物 32
表九、發酵製品的類型 33
表十、常被用作益生菌之乳酸菌株 40
表十一、通過衛生署健康食品認證與乳酸菌有關產品中synbiotics所含乳酸菌株
與益菌生 41
表十二、基礎培養基之組成 47
表十三、培養基中NNPL組成 48
表十四、香菇柄、杏鮑菇及金針菇基部之多醣產率 87
表十五、香菇、杏鮑菇及金針菇多醣以酚-硫酸法測定純度 88
表十六、以MRS培養基以及選擇性培養基計數乳酸益生菌 98
表十七、含L. acidophilus BCRC 20695之發酵乳於儲存期間其pH及存活菌數之
變化 99
表十八、含L. casei BCRC 14080之發酵乳於儲存期間其pH及存活菌數變化 100
表十九、含B. longum BCRC 14664之發酵乳於儲存期間其pH及存活菌數變化 101

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