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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/35147

Title: 靈芝複合菌株液態培養所得菌絲體與胞外多醣產量及其活性之探討
Production and bioactivity of mycelium and extracellular polysaccharide from mixed strains of Ganoderma lucidum using submerged fermentation
Authors: Hsun-Ming Chang
張勛銘
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: 靈芝;胞外多醣;複合菌株發酵;超氧化歧化酶;小鼠巨噬細胞;免疫調節
Ganoderma lucidum;extracellular polysaccharide;mixed strains fermentation;superoxide dismutase;RAW264.7 macrophage;immune-modulating
Date: 2013
Issue Date: 2013-10-07T02:50:57Z
Abstract: 本研究以本研究室自行研發之液態培養基 Gano medium 搖瓶震盪培養不同靈芝菌株,並行不同組合之複合菌株培養,希望利用此方式增加菌絲體及胞外多醣產量,並收集胞外多醣比較分子量分布情形,最後以小鼠巨噬細胞 RAW264.7 評估胞外多醣之免疫調節活性。首先由 8 株靈芝菌株 (Ganoderma lucidum) 中,篩選出菌絲體與胞外多醣產量較高的 5 株 (A、B、C、D、H),以此 5 株靈芝菌株進行 2 株、3 株、4 株菌與 5 株菌等 26 種組合複合菌株培養,篩選出菌絲體量最高組合 ACD (0.33 g/100 mL) 及胞外多醣產量最高組合 ABDH (0.344 g/L)。26 組複合菌株組合當中,以 DH 組合之菌絲體 SOD 活性為最高 (359.22 U/g),且僅有此組合超過 200 U/g,其次為 AH 組合 (197.26 U/g)。將菌體及胞外多醣產量較高的組合 ACD 及 ABDH 長時間培養 21 天,ACD 組合菌絲體含量於第 11 天時開始趨於平穩,第 21 天時菌絲體量為 0.49 g/100 mL,為第 7 天菌絲體量之 1.40 倍,而 ABDH 組合則是至第 17 天開始趨於平穩,其在第 21 天之菌絲體量 (0.40 g/100 mL),為第 7 天菌絲體量之 1.30 倍。ACD 與ABDH 組合第 21 天所得胞外多醣則分別是其第 7 天之 2.04 與 1.34 倍。ACD 及 ABDH 組合菌絲體之 SOD 活性均在第 9 天時達到高峰,分別為 76.53 及 110.44 U/g,之後逐漸下降,反觀培養液部份則是培養 21 天內均不超過 10 U/g。ACD 組合培養 21 天所得胞外多醣經過膠體過濾層析可得到分子量為 1557.8 kDa、552.7 kDa 及 5.2 kDa 之多醣,而 ABDH 則是得到 233.1 kDa 及 5.2 kDa 之多醣。以 RAW264.7 測試胞外多醣對細胞存活率、NO 分泌及吞噬活性之影響,兩種組合於濃度 10、40、100 與 400 g/mL 下均無顯著影響其存活率,且均以培養 21 天所得胞外多醣具有較佳抑制 NO 分泌之效果,在濃度 400 g/mL 下,可將 LPS 誘導產生之 NO 分別抑制 39.7% 及 40.1%。促吞噬活性則是以 ACD 組合培養 21 天及 ABDH 組合培養 7 天所得胞外多醣具有較佳促吞噬活性,在濃度 400 g/mL 下,可分別提升 3.38 及 2.85 倍之 細胞吞噬能力。
Five strains (A, B, C, D and H) of Ganoderma lucidum were chosen, based on data of mycelium amount and extracellular polysaccharide (EPS) production of 8 strains in medium at 30oC for 7 days. 26 combinations of mixed strains from these 5 srains. After measuring the mycelium amount and EPS production for 26 combinations of mixed strains from these 5 srains, ACD mixed cultures and ABDH mixed cultures had the highest mycelium amount (0.33 g/100 mL) and EPS production (0.344 g/L), respectively. DH mixed cultures produced the highest SOD activity in mycelium (359.22 U/g). The mycelium amount for ACD mixed cultures at 11th day reached the stationary phase and the mycelium amount at 21th day were 0.49 g/100 mL, which were 1.40 fold of that at 7th day. The mycelium amount for ABDH mixed cultures reached the stationary phase at 17th day and the mycelium amount at 21th day were 0.40 g/100 mL, which were 1.30 fold of that at 7th day. EPS amounts for ACD mixed strains and ABDH mixed cultures, were 2.04 and 1.34 folds of that at 7th day, respectively. Maximal mycelial SOD activity were observed at 9th day for ACD and ABDH mixed cultures, which were 76.53 and 110.44 U/g, while, SOD activity in culture filtrates of both mixed cultures were less than 10 U/g during 21 days of incubation. The EPS at 400 g/mL from ACD and ABDH mixed cultures for 21 days of incubation did not significantly affect cell viability; while the inhibition percentages for NO production in LPS-induced RAW264.7 cells by these two EPS at 400 g/mL were 39.7% and 40.1%, respectively. EPS samples at 400 g/mL from ACD mixed cultures for 21 days of incubation and from ABDH mixed cultures for 7 days of incubation increased the phagocytosis of RAW264.7 cells by 3.38 and 2.85 folds, respectively.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0010032031
http://ntour.ntou.edu.tw/handle/987654321/35147
Appears in Collections:[食品科學系] 博碩士論文

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