한국생물공학회:학술대회논문집
- 한국생물공학회 2001년도 추계학술발표대회
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- Pages.99-102
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- 2001
재조합 효모를 이용한 항혈전 단백질 히루딘 발효 생산공정의 최적화
초록
본 연구에서는 히루딘을 생산할 수 있는 재조합 S. cerevisiae 에서 ‘히루딘 유전자의 copy number 와 히루딘 발현양과의 관계를 규명하였으며 ,
Recombinant Saccharomyces cerevisiae strains harboring various copy numbers of hirudin gene were developed to study dependency of hirudin expression level on its gene copy number. A linear relationship between the copy number of hirudin expression cassette and hirudin expression level was observed up to 10 copies. A double <5-integration vector truncated wi 디 1 the unnecessary bacterial genes before yeast transformation showed a four-fold increase in transformation efficiency and a 1.3-fold enhancement in hirudin expression level compared with a single <5 system. Gratuitous hirudin expression strain was developed by disrupting the GALl gene of S. cerevisiae. Glucose that was fed in a limited manner effectively supported cell growth and hi겨din expression by the gratuitous strain. Effects of methanol concentrations on hirudin production in recombinant Hansenula polymorpha were investigated in continuous and fed-batch cultures. At a steady-state of continuous culture, an optimum methanol concentration of 1.7 g/L was determined at a dilution rate of 0.18
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