Effective production process of recombinant pharmaceutical protein with high purity oxygen고순도 산소를 이용한 재조합 의약용 단백질의 고효율 생산공정

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Microbial fermentation technology has been widely used as academic and industrial methods to treat wastewater and food-waste, produce useful proteins. In large scale aerobic fermentation processes, however, a high cell density culture was actually regarded as impossible method due to dissolved oxygen limitation and low competitive power. In this research, Hansenula polymorpha GOT7 was used as microorganism to investigate whether high purity oxygen affects secreting human serum albumin during batch and fed-batch fermentation processes. The optimum cultural conditions were established in advance through several basic flask cultures. Hansenula polymorpha GOT7 was shown best expression quantities at pH 6.0 and 37 ℃. Researches were progressed based on these results. The objectives of batch fermentation cultures were to confirm the mechanism of methanol oxidase promoter which regulates human serum albumin expression and investigate high purity oxygen effect. To achieve these two objectives, culture medium and supplied gases were changed at each case. The results showed that the most albumin (3.136 g/L) was secreted when YPG medium which contained glycerol were used. In case of YPD medium, human serum albumin was increased at 1.46 times, and 1.29 times at YPG medium. In fed-batch fermentation cultures, the effects of two important factors, carbon source feeding strategy and high purity oxygen supplying strategy, were investigated. In culture with YPG medium and high purity oxygen condition, specific growth rate, μ, was regulated manually to decrease slowly (gradual feeding) and induced with methanol. The results were 2.15 times increase (5.762 g/L) when compared with YPG medium and air condition (2.680 g/L). These increases were mainly due to the growth of cell mass. These experimental results showed the possibility of large scale microbial fermentation with high purity oxygen and high cell density cultures.
Advisors
Chang, Ho-Namresearcher장호남researcher
Description
한국과학기술원 : 생명화학공학과,
Publisher
한국과학기술원
Issue Date
2004
Identifier
238383/325007  / 020023382
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생명화학공학과, 2004.2, [ viii. 60 p. ]

Keywords

HIGH PURITY OXYGEN; RECOMBINANT PHARMACEUTICAL PROTEIN (ALBUMIN); YEAST; 효모; 고순도 산소; 재조합 의약용 단백질 (알부민)

URI
http://hdl.handle.net/10203/29826
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=238383&flag=dissertation
Appears in Collection
CBE-Theses_Master(석사논문)
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