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KMID : 0380020060210040260
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Korean Journal of Biotechnology and Bioengineering
2006 Volume.21 No. 4 p.260 ~ p.266
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Effect of Various Additives on the Production of Recombinant HBsAg during Methanol Induction in Pichia pastoris
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Lee Kyoung-Hoon
Lim Sang-Min
Kim Dong-Il
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Abstract
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Methanol induction conditions with various additives for the enhanced production of recombinant hepatitis B surface antigen (HBsAg) were investigated in Pichia pastoris, which can utilize methanol as a carbon source and produce recombinant proteins under the control of strong, tightly-regulated alcohol oxidase (AOX) promoter. The presence of non-methanol carbon sources such as glycerol and glucose fully repressed the expression of AOX promoter. Various additives were tested to improve the production of recombinant protein and it was found that sorbitol could be a good carbon source during methanol induction period. An optimized concentration of amino acid mixture enhanced the production of HBsAg significantly. Pluronic F-68, a non-ionic surfactant, also improved the production of HBsAg without inhibiting cell growth. Addition of oleic acid at 0.01% (v/v) during the induction period showed positive effect on the production of HBsAg. Finally, 1.2% (v/v) of trace salts enhanced the production of HBsAg 1.9 times compared to that of control culture.
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KEYWORD
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Pichia pastoris, alcohol oxidase promoter, hepatitis B surface antigen, pluronic F-68, oleic acid, trace salts
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