降温诱导改善猪α干扰素发酵生产性能和ATP再生效率-发酵工程专业论文.docxVIP

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降温诱导改善猪α干扰素发酵生产性能和ATP再生效率-发酵工程专业论文.docx

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Abstract Abstract wi也 a recombinant Pichia psatoris (IFN町PICZaA)部 mode 附盹伽 effects of environmental factors on porcine interferon-a (pIFN-α) concentration and antiviral activity were investigated in shake flask and 10L fermentor cultivations.ηle results in shake flask experiments showed 也at the influence of tempera阳re on pIFN-a expression w;ωmost significant.伽 the basis of shake flask expe血时时， the influence of temperature on pIFN-α production performance in 10L fermenter cultivation w;础 h时1er analized by simultaneously monitoring of fermentation process s阳te parame忧rs and detailed analysis of 由.e simplified Pichia psatoris methanol metabolic model.丁he main results ofthis 剑udyareωfollows: (1)To investigate the influence of environmenta1 factors on pIFN-a expression，也ree impo此ant environmental factors(temperature ，methanol concentration and pH) 扭扭duction ph邸e were selected for shake flask expe巾nent. By SDSanalysis，compared to 。induction at 30.C ，也e 恤tensity oftargeted pIFN-a band in 20.C cultivation w.础 significantly stronger at 72h after induction. The maximwn pIFN唱 concen甘ation of 0.19gIL in shake flask cultivation was obtained when induction at 20C ，pH 5.5 and methanol concentration of 20gIL. 。 )η1e Pichia pastoris high cell density cultivation experiment was carried out in 10L fermenter under different induction tempera阳res. The fermentation process state 阳ame忧m suchωspecific methanol consumption rate，。即gen uptake rate(OUR) ，C02 evolution rate (CER) and respirato叩 quotient(RQ) were on-line monitored by common instruments used in industrlal production ，and there were gre创 difference in 由.e above p町ameters when inducing at different temperatures.η1e cells metabolic activity and specific methanol consumption rate at 20.Cwas higher than 由创 when inducing at 30C.ηle simplified Pichia pωtoris methanol metabolism analysis indicated 也at the change of 始mpera阳re could lead to the change of metabolic flux ratio between energy generation pathway (pathway A)