深層培養松茸(Tricholoma matsutake) 之最適化培養液組成

Optimization of the medium components for Tricholoma matsutake submerged culture

張志豐(Chih-feng Chang)

指導教授 ： 李綉鈴

大同大學/工程學院/生物工程學系所/碩士(2004年)

摘要

松茸 (Tricholoma matsutake)又名松口蘑，是一種珍貴的食用菌，具有抗腫瘤及提高免疫力等生理活性。本研究乃在探討，於液態培養T. matsutake BCRC 36614中之培養液，包括碳源種類、胺基酸及銨態氮、有機氮peptone及yeast extract之添加，對松茸菌絲體及胞內多醣生產之影響，再利用反應曲面法找出各組成分之最適濃度。結果顯示，glucose、maltose、mannose、lactose及malt extract不僅促進T. matsutake BCRC 36614之生長，亦能促進胞內多醣之累積，其中以含有2 % glucose之基礎培養液培養T. matsutake BCRC 36614至四十天後，可獲得最大菌絲乾重，約為8.4 g/L，而胞內多醣含量亦最高，約為3.6 mg/100 mg CDW，產量約為30.1 mg/100 ml。L-arginine或L-glutamic acid之添加可促進T. matsutake BCRC 36614之生長及胞內多醣之累積；而ammonium tartrate或ammonium nitrate兩種銨態氮則有抑制效果。添加0.2 % L-arginine於培養液中，T. matsutake BCRC 36614之生長狀況最佳，培養至第四十天時，可獲得最大菌絲體乾重，約為9.9 g/L，為不添加者之1.2倍；而胞內多醣含量亦最高，約為7.4 mg/100 mg CDW，為不添加者之2倍，產量約為73.4 mg/100 ml。Peptone或yeast extract之添加，對T. matsutake BCRC 36614之生長及胞內多醣含量皆有顯著影響，以0.2 %為最適添加濃度。經由反應曲面實驗設計法得知，當培養基組成為2.25 % glucose、0.2 % peptone、0.22 % yeast extract、0.2 % L-arginine、0.05 % KH2PO4、0.05 % MgSO4•7H2O時，為培養T. matsutake BCRC 36614獲得最大胞內多醣含量之最適組成，約為7.3 mg/ 100 mg CDW。

關鍵字

反應曲面法 ； 深層培養 ； 真菌 ； 胞內多醣 ； 松茸

並列摘要

Tricholoma matsutake, also named matsutake, is a precious edible fungus which shows many pharmacological activities including anti-tumor effect and immunology enhancement. In this study, we investigated the effects of adding different carbon, amino acid, ammonium nitrogen and organic nitrogen (peptone and yeast extract) in T. matsutake BCRC 36614 submerged cultures on the growth and endopolysaccharide formation. Finally, we used response surface methodology to find the optimal concentration of each medium component. Our results showed that glucose, maltose, mannose, lactose and malt extract promoted not only the growth of T. matsutake BCRC 36614, but also the formation of endopolysaccharide. Among them, T. matsutake BCRC 36614 cultured in 2 % glucose-added basal medium after 40 days could result in maximal cell dry weight (8.4 g/l), endopolysaccharide content (3.6 mg/ 100mg CDW), and maximal endopolysaccharide production (30.1 mg/ 100 ml). Addition of L-arginine or L-glutamic acid promoted not only the growth of T. matsutake BCRC 36614, but also the formation of endopolysaccharide. However, two ammonium nitrogen, ammonium tartrate and ammonium nitrate, showed suppressive effects. The best growth of T. matsutake BCRC 36614 was obtained in 0.2 % L-arginine-added medium. In the 40-day culture, we obtained maximal cell dry weight (9.9 g/l) which was 1.2 times compared with the control group, maximal endopolysaccharide content (7.4 mg/ 100mg CDW) which was 2 times compared with its control group, and maximal production (73.4 mg/ 100 ml). Addition of peptone or yeast extract could significantly affect the growth and endopolysaccharide formation of T. matsutake BCRC 36614. The optimal concentrations of peptone and yeast extract were both 0.2 %. Using response surface methodology, we found the optimal medium components formula: 2.25 % glucose, 0.2 % peptone, 0.22 % yeast extract, 0.2 % L-arginine, 0.05 % KH2PO4, and 0.05 % MgSO4•7H2O. Cultured in the optimal medium, T. matsutake BCRC 36614 produced maximal endopolysaccharide content (7.3 mg/100 mg CDW).

並列關鍵字

endoplysaccharide ； fungi ； mushroom ； response surface methodology ； submerged culture ； Tricholoma matsutake

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