Development of high cell density recombinant Pichia Pastoris culture for human growth hormone production

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dc.contributor.authorLee, Yoeu Chek
dc.date.accessioned2014-11-03T02:09:24Z
dc.date.available2014-11-03T02:09:24Z
dc.date.issued2008
dc.descriptionMasteren_US
dc.description.abstractMethylotrophic yeast, Pichia pastoris has been used as an excellent host to produce more than 400 recombinant heterologous proteins of academic and industrial interests. Pichia pastoris strain used in this study was GS 115 2(5) with the phenotype of Mut+, which has been genetically modified to produce intracellular recombinant human growth hormone (rhGH). In this research, the development of fermentation protocols for high cell density culture of recombinant Pichia pastoris was carried out. Medium formulation and improved batch cultivation was investigated prior to fed batch fermentation. Various feeding strategies and methanol induction conditions on biomass and rHGH production were also studied. Ion exchange chromatography was performed to purify the rhGH produced. A protocol of the high cell density culture was developed which gave the highest dry cell weight of 120 g/L with the maximum specific growth rate of 0.133 h-1. A protocol that produces rHGH at the shortest time requirement was also obtained. It took only 39 hours from the inoculation to reach the rhGH production stage, which was almost 50% shorter time required compared to the preliminary 72 h. An enhanced induction condition for rhGH production was also developed where a yield of 2.4 mg of rhGH /L of lysate was achieved. A total of 349.5 μg of rhGH was purified from the total of 2418.8 μg of rhGH, which give the final yield of 14.5% and the purification factor of 1.5.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/203
dc.language.isoenen_US
dc.subjectBiological Scienceen_US
dc.subjectCell densityen_US
dc.subjectHormone productionen_US
dc.titleDevelopment of high cell density recombinant Pichia Pastoris culture for human growth hormone productionen_US
dc.typeThesisen_US
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