Biochemical Analysis of Baculovirus-insect Cell Interaction: I. Improved Recombinant ${\beta}-Galactosidase$ Production Using Medium Additives at AcNPV Infection of Insect Cells

Baculovirus-곤충세포 상호반응에 대한 생화학적 연구 -I. AcNPV의 곤충세포 감염시 배지 첨가물을 이용한 재조합 ${\beta}-galactosidase$ 생산 향상-

  • Lee, Ki-Woong (Department of Genetic Engineering, Kyung Hee University) ;
  • Kim, Tae-Yong (Department of Genetic Engineering, Kyung Hee University) ;
  • Chung, In-Sik (Department of Genetic Engineering, Kyung Hee University)
  • 이기웅 (경희대학교 자연과학대학 유전공학과) ;
  • 김태용 (경희대학교 자연과학대학 유전공학과) ;
  • 정인식 (경희대학교 자연과학대학 유전공학과)
  • Published : 1995.12.31

Abstract

The medium additives such as fatty acid, lipid, mannose, folic acid, $CaCl_2$ were examined to enhance recombinant ${\beta}-galactosidase\;({\beta}-gal)$ production in T-flask and air-lift bioreactor. The addition of each component. such as cholesterol, tocopherol, tricaprylin, mannose and folic acid at AcNPV infection of Tn5B1-4 cells enhanced ${\beta}-gal$ production, whereas the addition of $CaCl_2$ did not increase ${\beta}-gal$ production. The recombinant ${\beta}-gal$ production using the infection medium supplemented with a mixture of 0.34 mM cholesterol, 2.2 mM mannose and 0.045 M folic acid was enhanced 2 fold in an air-lift bioreactor, compared to the basal medium.

T-flask와 air-lift 생물반응기를 이용하여 Autographa californica nuclear polyhedrosis virus (AcNPV)의 Tn5B1-4 세포에의 감염시 fatty acid 및 lipid, mannose, folic acid, $CaCl_2$등의 배지 첨가물이 재조합 ${\beta}-galactosidase\;({\beta}-gal)$ 생산에 미치는 영향을 조사하였다. Cholesterol, tocopherol, tricaprylin 또는 mannose를 첨가하거나 folic acid를 보강첨가 했을 때 AcNPV의 재조합 ${\beta}-gal$ 생산은 향상되었으나 $CaCl_2$의 보강 첨가는 ${\beta}-gal$ 생산을 높이는데 효과적이지 못했다. Air-lift 생물반응기에서 0.34 mM cholesterol, 2.2 mM mannose 및 0.045 M folic acid를 보강 첨가한 배지를 이용하였을 때 재조합 ${\beta}-gal$ 생산은 basal medium에 비해 약 2배 정도의 증진 효과가 있었다.

Keywords