舞茸菌絲體醱酵及免疫調節功能探討

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標題:	舞茸菌絲體醱酵及免疫調節功能探討 Studies of Submerged Culture and Immunomodulatory Activity of Grifola frondosa TSRI01
作者:	鄭淑云 Shu-Yun Cheng
貢獻者:	吳明娟鄭作林嘉南藥理科技大學：生物科技研究所
關鍵字:	自然殺手細胞毒殺能力舞茸免疫調節作用吞噬作用 natural killer cytotoxicity CD11b immunomodulatory effect phagocytosis Grifola frondosa
日期:	2005
上傳時間:	2008-11-24 17:00:43 (UTC+8)
摘要:	前人研究發現，舞茸子實體中的多醣體具有抗腫瘤及免疫調節的功能探討因此已被使用做為抗腫瘤與調節免疫訴求的膳食補充品。本論文目的旨在探討舞茸(Grifola frondosa TSRI01)，以液態深層培養的最適醱酵條件，並建立人類全血免疫調節功能掍郊x，分析菌絲多醣萃取液及其分子量區分對人類全血免疫調節之幼纂C Grifola frondosa TSRI01 於5 公升醱酵槽進行攪拌、通氣培養，七天後獲得乾菌絲體10.37 g/L。接續放大至100 公升醱酵槽，經十天的攪拌、通氣培養，獲得乾菌絲體12.85 g/L。以過濾方法分離醱酵液及菌絲體，菌絲體再分別以熱水，冷鹼等逐步萃取，萃取液經酒精沉澱，獲得不同的區分，其中菌絲體熱水可溶區分，明顯誘導全血中TNF-ㄐBIL-6 和 IFN-γ等細胞激素的釋放。熱水可溶區分，再以不同濃度的酒精逐步沉澱，區分出不同分子量的多醣，以GPC測定多醣區分 I 及 II的分子量分佈，分別為43~140 kDa和13~38 kDa，區分 I 及II 對於誘導全血TNF-ㄐBIL-6 和 IFN-γ等細胞激素的釋放有顯著的效果，並提升全血中嗜中性顆粒球及單核球的吞噬能力。隨著多醣劑量的增加，多型核白血球(PMN)的CD11b的表現量也隨之增加。證明這些多醣區分可以有效活化補體受體CR3的表現，提升吞噬細胞的活性。同時多醣區分也具有促進人類自然殺手細胞毒殺K562人類骨髓腫瘤細胞的能力。本研究的體外試驗(in vitro)顯示，從舞茸TSRI01菌絲體所分離的多醣體區分，可以增加細胞免疫的調節能力，因此上述多醣成分可作為生物反應修飾劑(Biological response modifiers, BRM)。 Polysaccharides isolated from maitake (Grifola frondosa) fruiting bodyhave been used as dietary supplements due to their antitumor and immunomodulatory properties. The objective of this research was to optimize the conditions to produce the maximum amount of mycelia biomass by Grifola frondosa TSRI01 in submerged culture. This research was to produce and characterize the immunomodulatury functional polysaccharides of G. frondosa TSRI01.The optimal fermentation in a 5 L jar fermenter for 7days.The final biomass yield was 10.37g/L. G. frondosa was cultivated in a 100-L fermenter for 10 days. The final biomass yield was 12.85 g/L. The intracellular polysaccharide fraction, Fraction B, which was prepared by hot water extraction of mycelium followed by 4X ethanol precipitation, induced the release of TNF-ㄐBIL-6 and IFN-γ in human whole blood dose-dependently. Using the same strategy, we narrowed down the most potent fractions to Fractions I and II,which were prepared from the same hot water extract of mycelium by different ethanol concentration precipitation,respectively, and their active concentration was as low as 0.025 mg/mL. Chemical analysis revealed that Fractions I and II were composed of ~100% and ~92% of carbohydrates with very little protein contamination. Their molecular mass distributions were 43~140 and 13~38 kDa, respectively, as determined by GPC with refractive index detector.We also demonstrated that incubating human whole blood with 0.025 mg/mL of Fractions B, I and II enhanced phagocytic activities of PMN (polymorphonuclear neutrophils). The expression of CD11b, an early marker of PMN activation, was also up-regulated by these polysaccharides dose-dependently. This result suggested that complement receptor 3 (CR3) was primed by these fractions. In addition to activation of phagocytes, these bioactive fractions also increased human peripheral blood natural killer (NK) cell cytotoxicity. These results imply that the low-molecular-mass polysaccharides isolated from liquid-cultivated mycelium of G. frondosa TSRI01 can enhance innate immunity in vitro and therefore may serve as biological response modifiers (BRM).
關聯:	校內校外均不公開
顯示於類別:	[生物科技系(所)] 博碩士論文

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