篇名 | 利用紅豆芽中的麩胺酸脫羧酶生產γ-胺基丁酸之最適化生產條件探討 |
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卷期 | 56:5 |
並列篇名 | Optimized Production Conditions for Gama-amino Butyric Acid Using Glutamate Decarboxylase from Germinated Adzuki Bean (Vigna angularis) |
作者 | 陳巧馨 、 劉景文 、 江文德 |
頁次 | 128-136 |
關鍵字 | γ-胺基丁酸 、 麩胺酸脫羧酶 、 反應曲面法 、 紅豆 、 γ-amino butyric acid 、 Glutamate decarboxylase 、 Response surface methodology 、 Adzuki bean 、 Scopus 、 TSCI |
出刊日期 | 201810 |
DOI | 10.6578/TJACFS.201810_56(5).0003 |
本研究主要探討紅豆芽中麩胺酸脫羧酶 (GAD) 最適化生產γ-胺基丁酸 (GABA) 之條件。在固定酵素受質比值為0.0136、CaCl2 0.25 mM及反應時間1 h之下,以反應曲面法探討GAD最適化生產GABA之條件包括反應溫度、pH及pyridoxal phosphate (PLP) 濃度,其探討範圍分別為28.6~45.5℃ 、pH 6.32~9.68以及0.032~0.368 mM PLP,經由反應曲面回歸分析顯示,最適化生產條件為37.2℃ 、pH 7.6、0.208 mM PLP,可得最高的GABA產量326.7 mg/L,依照上述最適化條件進行驗證試驗,預測值在試驗值的95%信心度區間之內。接著利用生物反應器分離產物GABA並回收仍具有活性的GAD以持續轉換麩氨酸(Glu)生產GABA,回收利用GAD 2次後的生產率為197.3 mg GABA/mg protein是批次生產的2.89倍。儲存安定性試驗發現在4℃ 與-20℃ 保存半年均仍保有70%的GAD活性。
Response surface methodology was applied to evaluate the independent variables including reaction temperature 28.6~45.5oC, pH 6.32~9.68 and pyridoxal phosphate (PLP) concentration 0.032~0.368 mM under fixed enzyme to substrate ratio of 0.0136, 0.25 mM CaCl2 and reaction time 1 h. Based on response surface regression analysis procedure, the optimum reaction conditions for maximizing GABA production of 326.7 mg/L were 37.2oC, pH 7.6 and 0.208 mM PLP. The maximum predicted value could be achieved within 95% confidence interval of experimental values. Furthermore, bioreactor was employed to convert glutamate with GAD into GABA and to recycle the GAD. The productivity of bioreactor was 197.3 mg GABA/mg protein, which was 2.89 fold than that of batch reactor after reusing the GAD for twice. The result of storage stability test indicated that 70% GAD activity was maintained after half year storage under both 4oC and -20oC.