Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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Effect of Various Carbon Sources on the Production and Stabilization of hGM-CSF in Transgenic Plant Suspension Culture

형질전환된 식물세포에서 hGM-CSF 생산과 안정성에 대한 다양한 탄소원의 효과

  • Lee Jae-Hwa (Department of Bioscience and Biotechnology, Silla University)
  • Published : 2005.12.01
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Abstract

The effects of various carbon sources on the secretion of hGM-CSF, total protein and protease into the medium were investigated in transgenic tobacco cells. The dry cell weight (11.2 g/L) and wet cell weight (310.8 g/L) were highest at 30 g/L glucose after 5-day culture but, the dry cell weight (13.4 g/L) and wet cell weight (480 g/L) were highest at 30 g/L sucrose after 10-day culture. The total protein (110.3 mg/L), protease activity (3950 U/L) and total secreted hGM-CSF (56 mg/L) were highest at 30 g/L sucrose after 10-day culture. Stabilization of the total secreted protein and hGM-CSF in various carbon source concentrations was determined. Total secreted protein was most stabilized in the medium containing sucrose. However, the loss of the total protein was increased with the concentrations of high level in medium containing sorbitol, mannitol, fructose, and glucose. hGM-CSF was more stabilized in the medium containing sucrose than in the medium containing sorbitol, mannitol, fructose, glucose.

다양한 탄소원을 사용하였을 시, 세포성장에 미치는 영향은 배양 5일제 glucose를 사용한 배양세포에서 dry cell weight가 11.2 g/L, wet cell weight가 310.8 g/L로 가장 높았다. 이후 배양 10일차에는 오히려 sucrose를 탄소원으로 사용한 배양세포에서 dry cell weight가 13.4 g/L, wet cell weight가 480 g/L로 가장 높이 성장한 것을 확인할 수 있었다. Total secreted protein의 경우는 배양 10일차에 sucrose를 기본 탄소원으로 사용하였을 때 110.3 mg/L로 가장 높게 나왔다. Total secreted protease 역시, 배양 10일차에 sucrose를 기본 탄소원으로 사용한 배양씩에서 3950 U/L로 가장 많은 양이 측정되었다. 최종적으로 재조합된 단백질인 hGM-CSF의 생산량에 대한 측정결과, sucrose를 기본 탄소원로 사용한 배지에서 배양 10일차에 56 mg/L로 가장 높게 측정됨을 확인할 수 있었다. 이 외에 다양한 탄소원 농도변화에 따른 total secreted protein과 hGM-CSF의 안정성 평가를 확인해본 결과, total secreted protein의 경우, 비교대상으로 사용한 sucrose에서는 농도변화에도 큰 손실률을 보이지는 않았으나, sorbitol, mannitol, fructose, glucose의 경우에는 농도가 높아질수록 배양액내의 protein의 손실률이 증가 하였다. 총 분비된 hGM-CSF 역시 sucrose에 비해 sorbitol, mannitol, fructose, glucose에서는 분비된 hGM-CSF의 양이 크게 감소하는 것을 확인할 수 있었고, 탄소원의 농도가 높아질수록 hGM-CSF의 양이 감소하는 경향이 크게 나타났다.

Keywords

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