Wei Gong-Yuan, Li Yin, Du Guo-Cheng, Chen Jian
Key Lab of Industrial Biotechnology, Ministry of Education, Southern Yangtze University, Wuxi 214036, China.
Sheng Wu Gong Cheng Xue Bao. 2003 Nov;19(6):734-9.
The effects of dissolved oxygen (DO) and pH on glutathione batch fermentation by Candida utilis WSH-02-08 in a 7 liters stirred fermentor were investigated. It was shown that DO concentration is an important factor in glutathione production. With the initial glucose concentration of 30 g/L and a 5 L/min air flow rate, and the agitation rate less than 250 r/min, the DO concentration was not sufficient to satisfy the oxygen requirement during the fermentation. With an agitation rate of more than 300 r/min, the cell growth and glutathione production were enhanced significantly, with the dry cell mass and glutathione production were 20% and 25% higher than that at 200 r/min. When C. utilis WSH 02-08 was cultivated in a batch process without pH control, cell growth and glutathione production were inhibited, likely due to a dramatic decrease in the pH. Intracellular glutathione leakages were observed when the pH was 1.5 or less. To assess the effect of pH on glutathione production, six batch processes controlled at pH 4.0, 4.5, 5.0, 5.5, 6.0 and 6.5 were conducted. The yield was highest at pH 5.5, when the dry cell mass and yield were 27% and 95% respectively higher than fermentation without pH control. The maximal intracellular glutathione content (2.15 %) was also achieved at the pH. To improve our understandings on the effect of pH on the batch glutathione production, a modified Logistic equation and Luedeking-Piret equation were used to simulate cell growth and glutathione production, respectively, under different pH. Based on the parameters obtained by the nonlinear estimation, kinetic analysis was performed to elucidate the effect of pH on the batch glutathione production. The process controlled at pH 5.5 was proven to be the best due to the higher value of K(I) (substrate inhibitory constant in the Logistic equation), lower value of a and higher value of beta (slope and intercept in the Luedeking-Piret equation, respectively).
研究了溶解氧(DO)和pH值对产朊假丝酵母WSH-02-08在7升搅拌式发酵罐中进行谷胱甘肽分批发酵的影响。结果表明,溶解氧浓度是谷胱甘肽生产的一个重要因素。当初始葡萄糖浓度为30 g/L、空气流速为5 L/min且搅拌速率低于250 r/min时,溶解氧浓度不足以满足发酵过程中的需氧量。当搅拌速率高于300 r/min时,细胞生长和谷胱甘肽产量显著提高,干细胞质量和谷胱甘肽产量比200 r/min时分别高出20%和25%。当产朊假丝酵母WSH 02-08在无pH控制的分批过程中培养时,细胞生长和谷胱甘肽产量受到抑制,这可能是由于pH值急剧下降所致。当pH值为1.5或更低时,观察到细胞内谷胱甘肽泄漏。为了评估pH值对谷胱甘肽生产的影响,进行了六个控制在pH 4.0、4.5、5.0、5.5、6.0和6.5的分批过程。在pH 5.5时产量最高,此时干细胞质量和产量分别比无pH控制的发酵高出27%和95%。在该pH值下也实现了最大细胞内谷胱甘肽含量(2.15%)。为了加深我们对pH值对分批谷胱甘肽生产影响的理解,分别使用修正的Logistic方程和Luedeking-Piret方程来模拟不同pH值下的细胞生长和谷胱甘肽生产。基于通过非线性估计获得的参数,进行动力学分析以阐明pH值对分批谷胱甘肽生产的影响。由于Logistic方程中的底物抑制常数K(I)值较高、a值较低以及Luedeking-Piret方程中的斜率和截距β值较高,证明pH 5.5控制的过程是最佳的。