甘油及甲醇补料策略对毕赤酵母表达猪α-干扰素的影响

本文研究了甘油及甲醇补料策略对重组毕赤酵母细胞生长及猪α-干扰素(pIFN-α)表达的影响。结果表明, 甘油采取不同的补料策略, 以及甲醇浓度控制于不同水平时, 细胞生长速度不同, pIFN-α抗病毒活性水平也明显不同。高密度发酵阶段, 甘油采用指数方式流加控制时, 相较于40 g/L/h、10 g/L/h, 恒速流加组的细胞浓度最先达最高水平132 g/L, 后一直保持稳定, 且发酵液中几乎无残留甘油; 与之相应的pIFN-α抗病毒活性水平最高。诱导表达阶段, 甲醇浓度须控制并恒定于12 g/L时, pIFN-α抗病毒活性最高水平能达到5.95×106 IU/mL, 而当甲醇浓度稳定于3.5 g/L、16.0 g/L、或者浓度波动较大(10.2～13.8 g/L)时, pIFN-α抗病毒活性均远低于最高水平。

Abstract

In this paper, effects of the feeding strategy of glycerol and methanol on Pichia pastoris growth and pIFN-α expression were investigated. The results showed that cell growth rate and antiviral activity of pIFN-α were both different under different glycerol feeding rates and methanol concentrations. In the exponential phase, which was fed by exponential method with glycerol, cell concentration reached the highest yield of 132 g/L more quickly than fed by glycerol with constant flow as 40 g/L/h and 10 g/L/h, the cell concentration also remained stable and there was almost no glycerol left in the broth; And the corresponding antiviral activity of pIFN-α was the highest to the highest cell concentration. During the induced expression phase, only when the methanol concentration was stabilized at 12 g/L, the antiviral activity of pIFN-α could reached 5.95×106 IU/mL. While the methanol concentration was at 3.5 g/L or 16.0 g/L, or ranged from 10.2 g/L to 13.8 g/L, antiviral activity of pIFN-α was much less than the highest production of 5.95×106 IU/mL.

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王丙莲, 冯东, 梁晓辉, 史建国, 刘仲汇. 甘油及甲醇补料策略对毕赤酵母表达猪α-干扰素的影响[J]. 激光生物学报, 2016, 25(6): 553. WANG Binglian, FENG Dong, LIANG Xiaohui, SHI Jianguo, LIU Zhonghui. Effects of Glycerol and Methanol Supplementation on Porcine Interferon α (PIFN-α) Expression of Pichia Pastoris[J]. Acta Laser Biology Sinica, 2016, 25(6): 553.

甘油及甲醇补料策略对毕赤酵母表达猪α-干扰素的影响

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