本研究以樟芝Antrodia camphorata BCRC 35396於攪拌式發酵槽培養下，探討不同通氣量對於樟芝發酵生產脂解酵素的影響，並將最適條件之發酵液進行酵素的分離純化。由實驗結果顯示，0.3 vvm的最大脂解酵素活性為1.151 U/ml，而1.0 vvm的最大酵素活性為2.588 U/ml，大約提升了2.25倍，所以通氣量的增加有利於脂解酵素活性的提升。而YL/P(specific activity)顯示，0.8 vvm有較高的比活性(29.493 U/mg)。在蛋白質的生成方面，0.3 vvm的最大蛋白質含量為0.061 mg/ml，而1.0 vvm的最大蛋白質含量為0.120 mg/ml，大約提升1.97倍，所以提升通氣量對於蛋白質含量有提高的效果。 脂解酵素的純化步驟是將發酵澄清液利用冷凍乾燥再經由DEAE Sepharose陰離子交換管柱層析分離純化。經過冷凍乾燥處理的脂解酵素活性及比活性分別為341.6 U及29.19 U/mg。而通過陰離子交換管柱層析後，純化倍率為4.50倍，回收率為8.28％。最後由SDS-PAGE電泳分析，發現最後純化的脂解酵素分子量是60.0 kDa。 This work studied the production and purification of lipase fermented from Antrodia camphorata BCRC 35396 by aeration rate in the stired tank. Shown by the experimental result, the more aeration rate can enhance the lipase activity. The biggest lipase activity of 1.0 vvm was 2.25 times higher than that of 0.3 vvm. The specific activity YL/P show the 0.8 vvm has the higher quality than other aeration rate. The biggest protein of 1.0 vvm was 1.97 times higher than one of 0.3 vvm. Lipase fermented from Antrodia camphorata BCRC 35396 was purified by lyophilization as concentration, follow by anion exchange chromatography(DEAE-Sepharose). The specific activity of the purified enzyme was 4.5 times higher than that of the crude broth after lyophilization. The lipase had a molecular weight of 60.0 kDa when examined by SDS-polyacrylamide gel electrophoresis.
