Application of a radial-flow bioreactor in the production of beta1,3-N-acetylglucosaminyltransferase-2 fused with GFPuv using stably transformed insect cell lines.
An RFB (radial-flow bioreactor) with a reactor volume of 5 ml was applied to produce human beta3GnT (beta1,3-N-acetylglucosaminyltransferase) using two stably transformed insect cell lines. When air was supplied to the RFB, cell growth stopped at 4 days of culture and beta3GnT was not detected. However, with a supply of pure oxygen, the cell concentration, assumed from glucose consumption, increased by 1.3x10(7) cells/ml. Insect cells attached to poly(vinyl alcohol) matrixes packed in the RFB and grew confluently; 5.6 m-units/ml beta3GnT was produced under the conditions of pure oxygen supply and addition of glucose and glutamine. This RFB was first applied in beta3GnT production using stably transformed insect cells. The amount of beta3GnT production in only a 5 ml-scale RFB was comparable with that of a 100 ml shaking flask culture.
Kwon
Mi Sun
MS
Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, 836 Ohya, Shizuoka 422-8529, Japan.
Kato
Tatsuya
T
Dojima
Takashi
T
Park
Enoch Y
EY
D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D002460:Cell Line; D049109:Cell Proliferation; D004867:Equipment Design; D019544:Equipment Failure Analysis; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000528:radiation effects; D006801:Humans; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods; D009036:Moths / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D017351:N-Acetylglucosaminyltransferases / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014170:Transformation, Genetic; D014466:Ultraviolet Rays
D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D017351:N-Acetylglucosaminyltransferases; C428132:B3GNT2 protein, human