[Constitutive expression of human angiostatin in Pichia pastoris using glycerol as only carbon source].
Sheng Wu Gong Cheng Xue Bao
; 23(5): 902-6, 2007 Sep.
Article
em Zh
| MEDLINE
| ID: mdl-18051873
Carbon source plays an important role in the constitutive expression of foreign proteins in Pichia pastoris. In present study, glucose , glycerol , methanol and oil acid, was used respectively as the only carbon source to constitutively express hAS in Pichia pastoris GS115 (pGAP9K-AS)in shaking flask. The result shows that oleic acid is the best (163 mg/L) compared with glycerol (83mg/L), glucose (76 mg/L)and methanol (57 mg/L). Since oleic acid is insoluble in water, glycerol was used as the carbon source in the high-density cell culture of GS115 (pGAP9K-AS) in a 30 liter bioreactor and 169 mg/L of angiostatin was obtained after 48h of culture. The expressed angiostatin is immunologically active as shown by Western blotting. The recombinant hAS inhibits bFGF induced CAM angiogenesis and suppresses the growth of B16 melanoma in C57BL/6J mice. The tumor inhibition rate is 90% after 12 days of treatment. Statistics analysis revealed that the tumor volume difference of mice between the hAS group and PBS group is prominent (P < 0.01).
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Pichia
/
Reatores Biológicos
/
Angiostatinas
/
Glicerol
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Sheng Wu Gong Cheng Xue Bao
Ano de publicação:
2007
Tipo de documento:
Article