Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, People's Republic of China.
Bioprocess Biosyst Eng. 2012 Sep;35(7):1125-36. doi: 10.1007/s00449-012-0697-1. Epub 2012 Feb 15.
The production of porcine interferon-α (pIFN-α) by Pichia pastoris was largely enhanced when adopting sorbitol/methanol co-feeding induction strategy at 30 °C in a 10-L fermentor. Analysis of energy regeneration pattern and carbon metabolism revealed that major energy metabolism energizing pIFN-α synthesis shifted from formaldehyde dissimilatory energy metabolism pathway to TCA cycle under the methanol/sorbitol co-feeding induction strategy. The sorbitol/methanol co-feeding induction strategy weakened formaldehyde dissimilatory pathway and repressed the accumulation of toxic metabolite-formaldehyde, reduced theoretical oxygen consumption rate and oxygen supply requirement, and increased energy/methanol utilization efficiency so that more methanol could be effectively used for pIFN-α synthesis. As a result, pIFN-α antiviral activity reached a highest level of 1.8 × 10(7) IU/mL which was about 10- to 200-folds of those obtained under pure methanol induction at 20 and 30 °C, respectively.
当在 10 升发酵罐中于 30°C 下采用山梨醇/甲醇共补料诱导策略时，毕赤酵母生产猪干扰素-α（pIFN-α）的产量大大提高。能量再生模式和碳代谢分析表明，在甲醇/山梨醇共补料诱导策略下，为 pIFN-α 合成提供动力的主要能量代谢从甲醛异化能量代谢途径转移到 TCA 循环。山梨醇/甲醇共补料诱导策略削弱了甲醛异化途径，抑制了有毒代谢物甲醛的积累，降低了理论耗氧率和供氧需求，提高了能量/甲醇利用效率，从而可以更有效地利用甲醇用于 pIFN-α 的合成。结果，pIFN-α 的抗病毒活性达到了 1.8×10(7)IU/mL 的最高水平，分别比在 20 和 30°C 下纯甲醇诱导时获得的水平高出 10-200 倍。
