杏鮑菇液態菌種培養之研究

Studies of liquid spawn culture for Pleurotus eryngii

陳志豪(Chih-Hao Chen)

指導教授 ： 詹効松 ； 石信德

朝陽科技大學/理工學院/生化科技研究所/碩士(2011年)

摘要

中文摘要
本研究目的是以杏鮑菇ESPS品系，開發液態菌種發酵技術，生產菌絲體以利於商業化栽培。利用CS基礎培養基分別添加數種單糖、雙糖與有機添加物，比較其對杏鮑菇菌絲體生長之影響，試驗結果顯示菌絲經15,000 rpm轉速均質化後以10 %接種量進行培養，在單因子試驗中之碳源種類試驗結果以蔗糖與葡萄糖產生的菌絲量較高；添加有機氮源部份之結果可發現酵母萃取物與蛋白腖對菌絲體生長的影響極為顯著。將上述4種因子進行回應曲面法試驗 (Response Surface Methodology, RSM)，在一階迴歸方程式中顯示添加此4種因子對菌絲體生質量皆有正面影響，其中以酵母萃取物添加最為顯著，而在陡升路徑試驗中菌絲體生質量在試驗第4組時達到最高，則得到一新的中心混成試驗原點 (蔗糖2.48 %、葡萄糖1.11 %、酵母萃取物2.18 % 和蛋白腖0.68 %)，將此中心混成原點進行中心混成試驗。所得實驗數據結果以多元迴歸技巧進行二階模式理論契合，由回應曲面法求得杏鮑菇菌絲體產生最大生質量之培養基組成，其生質量產量之理論預測值為9.48 g/L，經搖瓶試驗驗證結果為10.19 g/L，此顯示回應曲面模式之可靠性。相較於CS培養基，最適化培養基生產菌絲體之生質量為其10.7倍。將最適化培養基擴大至6公升攪拌式生物反應器培養8天，將培養之液態菌種接種於太空包，結果顯示所有液態菌種之接種條件走菌速度均高於木屑菌種，子實體產量也比木屑菌種高，其中以接種條件為20 mL之液態菌種菌絲長滿太空包平均時間為25天，而木屑菌種平均時間則為45天；子實體產量可達331.67±7.55 g/bag為木屑菌種的 (276.67±6.75 g/bag) 1.2倍；而生物效率為94.22 %對照組則為78.60 %。

關鍵字

太空包 ； 攪拌式生物反應器 ； 杏鮑菇 ； 液態菌種培養 ； 回應曲面試驗設計法

並列摘要

Abstract‏
Pleurotus eryngii is an edible mushroom it has a thick, meaty white stem and a small tan cap. The mushroom has a good shelf life but an effective cultivation method is needed to enhance the biomass of this fungus for commercialization. Hence, the present work describes the use of different additives (sugars and nitrogen source) and treatments to enhance the biomass of commercially used P. eryngii ESPS strain for liquid spawn. The CS medium was used as basal medium and mycelium were grown in submerged culture conditions. The initial inoculums were obtained from the mycelia harvested from the petri dish and homogenized at 15,000 rpm. Approximately, 10 % (v/v) homogenized mycelia was used as initial inoculums and transferred to the 250 mL Erlenmeyer flask containing 50 mL of liquid CS medium. Effect of different monosaccharides, disaccharides and organic nitrogen source were examined one by one on mycelium growth. The significant growth response was observed with different carbon source (sucrose and glucose) and nitrogen source (yeast extract and peptone). Response Surface Methodology (RSM), was used to investigate the effect of these additives on mycelia growth. With the goal to achieve maximum mycelium production four components were tested in the process of media optimization. According to central component design and mathematical calculations the optimal concentration of sucrose, glucose, yeast extract and peptone were 24.8 g/L, 11.1 g/L, 21.8 g/L and 6.8 g/L respectively. The prediction of maximum mycelium production was 9.48 g/L, which coincides with the shaker-flask mycelium production 10.19 g/L. Based on results obtained from RSM, the experiment was extended to 6-L stirred tank reactor (STR) and mycelium were grown for eight days. About, 20 mL of this inoculum was poured as seed in bag cultivation as well as in sawdust. The high growth rate and fruiting body production were obtained in bag cultivation than the sawdust cultivation. An increase of 1.2 fold of the fruiting body production was obtained with bag cultivation (331.67 ± 7.55 g) compared to sawdust spawn (276.67 ± 6.75 g). The efficiency of cultivation was increased to 94.22% in bag cultivation compared to 78.60% in the control group. The present investigation on media optimization can be used for higher biomass production of P. eryngii ESPS strain and may be implemented to other Pleurotus strains.

並列關鍵字

Pleurotus eryngii ； liquid spawn culture ； stirred tank reactor (STR) ； Response Surface Methodology (RSM) ； bag cultivation.

參考文獻

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