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| High density fermentation control of Pichia pastoris |
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Abstract One of the important strategies for improving recombinant protein expression in Pichia pastoris is optimizing control during the high density fermentation process. Some fermentation control has been used and got obviously higher expression level: optimization of basal salt medium composition; using two stage pH and two stage temperature control during cell growth and induction stage; appropriately increasing dissolved oxygen; selecting optimal cell density prior to induction (CWPTI) and specific growth rate prior to induction (μPTI), using limit amount of initial glycerolin the basic media and feeding it during the fermentation with exponential fedbatch Model, selecting appropriate methanol feeding strategy by methanollimited feeding batch (MLFB), oxygenlimited feeding batch (OLFB), methanolunlimited feeding batch (MNLFB) or temperaturelimited feeding batch (TLFB) . Some research suggested that the expression level of the recombinant protein increase 9 times after optimizing the fermentation control.
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Received: 16 September 2008
Published: 06 May 2009
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| ［1］ Daly R，Hearn M T W. Expression of heterologous proteins in Pichia pastoris: a useful experimental tool in protein engineering and production. J Mol Recognit,2005,18 (2):119~138
［2］ Hasslacher M，Schall M，Hayn M，et al. Highlevel intracellular expression of hydroxynitrile lyase from the tropical rubber tree Hevea brasiliensis in microbial hosts. Protein Expres Purif，1997，11 (1)：61~71
［3］ Xiong A S，Yao Q H，Peng R H，et al. High level expression of a synthetic gene encoding Peniophora lycii phytase in methylotrophic yeast Pichia pastoris. Appl Microbiol Biotechnol，2006，72 (5)：1039~1047
［4］ Zhang J G，Wang X D，Zheng Y，et al. Enhancing yield of Sadenosylmethionine in Pichia pastoris by controlling NH4+ concentration. Bioprocess Biosyst Eng，2008，31 (2)：63~67
［5］ Huang H Q，Yang P L，Luo H Y，et al. Highlevel expression of a truncated 1,31,4βDglucanase from inFibrobacter succinogenes Pichia pastoris by optimization of codons and fermentation. Appl Microbiol Biotechnol，2008，78：95~103
［6］ 洒荣波，石贵阳，王正祥. 基因工程菌Pichia pastoris 密度培养条件的摇瓶研究. 食品研究与开发，2005，26 (2)：52~56
Sa R B, Shi G Y, Wang Z X. Food Research and Developent，2005，26 (2)：52~56
［7］ Reverter D，Ventura S，Villegas V，et al. Overexpression of human procarboxypeptidase A2 in Pichia pastoris and detailed characterization of its activation pathway. The Journal of Biological Chemistry，1998，273 (6)：3535~3541
［8］ 周祥山，范卫民，张元兴. 不同甲醇流加策略对重组毕赤酵母高密度发酵生产水蛭素的影响. 生物工程学报，2002，18 (3)：348~351
Zhou X S, Fan W M, Zhang Y X. Chinese Journal of Biotechnology，2002，18 (3)：348~351
［9］ 谢静莉，周庆玮，杜鹏，et al. 重组巴斯德毕赤酵母高密度培养中铵离子浓度的影响. 华东理工大学学报，2004，30 (6)：723~726
Xie J L, Zhou Q W, Du P, et al. Jouranl of East China University of Science and Technology，2004，30 (6)：723~726
［10］ Yang J Z，Zhou X S，Zhang Y X. Improvement of recombinant hirudin production by controlling NH4+ concentration in Pichia pastoris fermentation. Biotechnology Letters，2004，26 (12)：1013~1017
［11］ Surribas A，Stahn R，Montesinos J L，et al. Production of a Rhizopus oryzae lipase from Pichia pastoris using alternative operational strategies. Journal of Biotechnology，2007，130 (3)：291~299
［12］ Zhao H L，Xue C，Wang Y，et al. Increasing the cell viability and heterologous protein expression of Pichia pastoris mutant deficient in PMR1 gene by culture condition optimization. Appl Microbiol Biotechnol，2008，81(2)：235~241
［13］ 李洪淼，王红宁，许钦坤. 毕赤酵母高密度发酵研究进展. 生物技术通讯，2005，16 (2)：210~212
Li H M, Wang H N, Xu Q K. Letters in Biotechnology，2005，16 (2)：210~212
［14］ Xiao A F，Zhou X S，Zhou L，et al. Improvement of cell viability and hirudin production by ascorbic acid in Pichia pastoris fermentation. Appl Microbiol Biotechnol，2006，72 (4)：837~844
［15］ Ayed A，Rabhi I，Dellagi K，et al. High level production and puri?cation of human interferon α2b in high cell density culture of Pichia pastoris. Enzyme Microb Technol，2008，42：173~180
［16］ 孙战胜，陈劲春. 重组人血清白蛋白在毕赤酵母表达中的降解控制. 北京化工大学学报 ，2004，31 (4)：9~15
Sun Z S, Sheng J C. Journal of Beijing University of Chemical Technology，2004，31 (4)：9~15
［17］ 姚钰舜，储炬，杭海峰，et al. 培养条件对重组毕赤酵母高密度表达猪胰岛素前体的影响. 华东理工大学学报（自然科学版），2006，32 (4)：397~401
Yao Y S, Chu J, Hang H F, et al. Journal of East China University of Science and Technology(Natural Science Edition)，2006，32 (4)：397~401
［18］ 史琪琪，郝玉有，吴康华，et al. 诱导相温度对毕赤酵母表达重组人复合α干扰素聚合的影响. 生物工程学报，2006，22 (2)：311~315
Shi Q Q, Hao Y Y, Wu K H, et al. Chinese Journal of Biotechnology，2006，22 (2)：311~315
［19］ Jahic M，Wallberg F，Bollok M，et al. Temperature limited fedbatch technique for control of proteolysis in Pichia pastoris bioreactor cultures. Microbial Cell Factories，2003，2 (1)：6
［20］ 方曙光，储炬，黄立，et al. 温度对巴斯德毕赤酵母表达瑞替普酶( reteplase ) 的影响.工业微生物，2007，37 (4)：11~15
Fang S G, Chu J, Huang L, et al. Industrial Microbiology，2007，37 (4)：11~15
［21］ Charoenrat T，KetudatCairns M，StendahlAndersen H，et al. Oxygenlimited fedbatch process: an alternative control for Pichia pastoris recombinant protein processes. Bioprocess Biosyst Eng，2005，27 (6)：399~406
［22］ Chen H X，Chu J，Zhang S L，et al. Intracellular expression of Vitreoscilla hemoglobin improves Sadenosylmethionine production in a recombinant Pichia pastoris. Appl Microbiol Biotechnol，2007，74 (6)：1205~1212
［23］ Wang Y，Wang Z H，Du G C，et al. Enhancement of alkaline polygalacturonate lyase production in recombinant Pichia pastoris according to the ratio of methanol to cell concentration. Bioresource Technol，2009，100 (3)：1343~1349
［24］ Wei C，Zhou X S，Zhang Y X. Improving intracellular production of recombinant protein in Pichia pastoris using an optimized preinduction glycerolfeeding scheme. Appl Microbiol Biotechnol，2008，78 (2)：257~264
［25］ Zhang J G，Wang X D，Su E Z，et al. A new fermentation strategy for Sadenosylmethionine production in recombinant Pichia pastoris. Biochem Eng J，2008，41：74~78
［26］ Sinha J，Inan M，Fanders S，et al. Cell bank characterization and fermentation optimization for production of recombinant heavy chain Cterminal fragment of botulinum neurotoxin serotype E (rBoNTE(H(c)): antigen E) by Pichia pastoris. Journal of Biotechnology，2007，127 (3)：462~474
［27］ Bahrami A，Shojaosadati S A，Khalilzadeh R，et al. Twostage glycerol feeding for enhancement of recombinant hGCSF production in a fedbatch culture of Pichia pastoris. Biotechnology letters，2008，30 (6)：1081~1085
［28］ 宋留君，周长林，窦洁，et al. 甘油对毕赤酵母高密度发酵表达重组水蛭素II的影响. 中国药科大学学报，2006，37 (4)：371~374
Song L J, Zhou C L, Dou J, et al. Journal of China Pharmaceutical University，2006，37 (4)：371~374
［29］ Khatri N K，Hoffmann F. Impact of methanol concentration on secreted protein production in oxygenlimited cultures of recombinant Pichia pastoris. Biotechnology and Bioengineering，2006，93 (5)：871~879
［30］ Khatri N K，Hoffmann F. Oxygenlimited control of methanol uptake for improved production of a singlechain antibody fragment with recombinant Pichia pastoris. Appl Microbiol Biotechnol，2006，72 (3)：492~498
［31］ Trinh L B，Phue J N，Shiloach J. Effect of methanol feeding strategies on production and yield of recombinant mouse endostatin from Pichia pastoris. Biotechnology and Bioengineering，2003，82 (4)：438~444
［32］ Hu X Q，Chu J，Zhang S L，et al. A novel feeding strategy during the production phase for enhancing the enzymatic synthesis of SadenosylLmethionine by methylotrophic Pichia pastoris. Enzyme Microb Technol，2007，40：669~674
［33］ Jungo C，Schenk J，Pasquier M，et al. A quantitative analysis of the benefits of mixed feeds of sorbitol and methanol for the production of recombinant avidin with Pichia pastoris. Journal of Biotechnology，2007，131 (1)：57~66
［34］ Jungo C，Marison I，von Stockar U. Mixed feeds of glycerol and methanol can improve the performance of Pichia pastoris cultures: A quantitative study based on concentration gradients in transient continuous cultures. Journal of biotechnology，2007，128 (4)：824~837
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