Effect of Methanol Concentration on the Production of Human β_2-Glycoprotein I Domain V by a Recombinant Pichia pastoris : A Simple System for the Control of Methanol Concentration Using a Semiconductor Gas Sensor

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The methylotrophic yeast Pichia pastoris is one of the best for the production of foreign proteins because of the presence of the strong AOX1 promoter induced by methanol. Methanol feeding during the production phase of the foreign proteins is important because methanol not only induces protein production but also provides energy source for the host cells. Excess methanol inhibits the growth of host cells, while an insufficient amount of energy source and/or methanol starvation lead to poor growth and production. We constructed a simple methanol control system consisting of a semiconductor gas sensor and a relay. Using this system, we studies the effect of methanol concentration on the production of a model foreign protein, human β_2-glycoprotein I domain V. The methanol concentrations were kept constant at 1.5, 10, 17, or 31g・l^<-1>(±5%) during the production phase. Although the specific rates of growth and methanol consumption decreased with increase in the methanol concentration, the specific production rates increased, indicating that the energy for the production competed with that for cell growth. Accordingly, we provided glycerol as an extra energy source during the production phase, with the result that the specific production rate increased two times. Our simple and inexpensive system will help bioengineering studies on the production of recombinant proteins in P.pastoris, the growth and production of objective proteins in which are dependent on the methanol concentration.
社団法人日本生物工学会の論文
1998-11-25

Effect of Methanol Concentration on the Production of Human β_2-Glycoprotein I Domain V by a Recombinant Pichia pastoris : A Simple System for the Control of Methanol Concentration Using a Semiconductor Gas Sensor

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