甘油及甲醇补料策略对毕赤酵母表达猪α-干扰素的影响  

Effects of Glycerol and Methanol Supplementation on Porcine Interferon α( PIFN-α) Expression of Pichia Pastoris

作　　者：王丙莲[1,2] 冯东[1,2] 梁晓辉[3] 史建国[1,2] 刘仲汇[1,2] WANG Binglian FENG Dong LIANG Xiaohui SHI Jianguo LIU Zhonghui(l. Biology Institute of Shandong Academy of Sciences, Jinan 250014, Shandong, China Key Laboratory for Biosensors of Shandong Province, Jinan, Shandong, China Energy Research Institute of Shandong Academy of Sciences, Jinan 250014, Shandong, China)

机构地区：[1]山东省科学院生物研究所,山东济南250014 [2]山东省生物传感器重点实验室,山东济南250014 [3]山东省科学院能源研究所,山东济南250014

出　　处：《激光生物学报》2016年第6期553-558,共6页Acta Laser Biology Sinica

基　　金：山东省重点研发项目(2015GSF121047);山东省重点研发项目(2015GSF117034)

摘　　要：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（p IFN-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,p IFN-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p IFN-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,p IFN-α抗病毒活性最高水平能达到5.95×10^6IU/m L,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,p IFN-α抗病毒活性均远低于最高水平。In this paper,effects of the feeding strategy of glycerol and methanol on Pichia pastoris growth and p IFN-αexpression were investigated. The results showed that cell growth rate and antiviral activity of p IFN-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 p IFN-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 p IFN-α could reached 5. 95 × 10^6 IU/m L. 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 p IFN-α was much less than the highest production of 5. 95 × 10^6 IU/m L.

关 键 词：甘油 甲醇 补料控制 猪α-干扰素 

分 类 号：TQ920[轻工技术与工程—发酵工程]