Li Xiao, Huang Cong, Xu Chao-Qun, Tan Ya-Li, Luo Yu-Di, Zou Kun, Li Jian-Hua, Deng Zhang-Shuang, Zheng Zhong, Ye Han, Zhang Xiao-Long, Zheng Nian
Hubei Key Laboratory of Natural Products Research and Development, College of Biological and Pharmaceutical Sciences, China Three Gorges University, Yichang, China.
Angel Yeast Co., Ltd., Yichang, China.
Biotechnol Appl Biochem. 2019 May;66(3):389-397. doi: 10.1002/bab.1735. Epub 2019 Feb 12.
The high cell density culture of baker's yeast FX-2 was investigated in a 50 L(A) automatic bioreactor. Herein, it was found firstly that the Crabtree effect clearly existed in batch fermentation with higher glucose content, then the critical initial glucose content range (≤2.00 g L ) was reasonably ascertained to effectively avoid Crabtree effect. In the next fed-batch fermentations with different strategies, the second strategy (maintain ethanol concentration lower than 0.10% and pH around 4.80) was confirmed to be more beneficial to yeast growth than the first strategy (keep reducing sugar not more than 2.00 g L and control steady Carbon/Nitrogen ratio 3.05:1.00). After that, one optimal control strategy (maintain pH around 4.80 and keep respiratory quotient in the range of 0.90-1.00) was constructed to further enhance cell yield. Under an optimal control strategy, four schemes with the aim of achieving pH-stat were compared, and yeast extract instead of other alkaline materials was selected as a better regulator. As a result, 148.37 g L dry cell weight, 38.25 × 10 mL living cells, and 8.24 g L h productivity were harvested, which respectively elevated 23.74%, 135.38%, and 24.47% compared to that obtained under the traditional scheme (regulate pH with ammonia); meanwhile, the maximum oxygen uptake rate and carbon dioxide excretion rate were both more than 250.00 mmol L min .
在50L(A)自动生物反应器中对面包酵母FX-2进行了高细胞密度培养研究。在此发现,首先在葡萄糖含量较高的分批发酵中明显存在克勒勃屈利效应,然后合理确定了临界初始葡萄糖含量范围(≤2.00g/L)以有效避免克勒勃屈利效应。在接下来不同策略的补料分批发酵中,证实第二种策略(保持乙醇浓度低于0.10%且pH值在4.80左右)比第一种策略(保持还原糖不超过2.00g/L并控制稳定的碳氮比为3.05:1.00)更有利于酵母生长。此后,构建了一种最优控制策略(保持pH值在4.80左右并使呼吸商在0.90 - 1.00范围内)以进一步提高细胞产量。在最优控制策略下,比较了四种旨在实现恒pH的方案,选择酵母提取物而非其他碱性物质作为更好的调节剂。结果,收获了148.37g/L干细胞重量、38.25×10个活细胞以及8.24g/L·h的生产率,与传统方案(用氨调节pH)相比分别提高了23.74%、135.38%和24.47%;同时,最大氧摄取率和二氧化碳排泄率均超过250.00mmol/L·min。