Fig. 3

Intracellular expression of pGH in Pichia pastoris X33 after gene optimization. a SDS–PAGE analysis of soluble and insoluble protein fractions of Pichia pastoris X33 expressing optimized pGH under the control of AOX1 promoter after induction at 30 °C. Lanes 1 and 6, non optimized pGH gene for the expression of soluble pGH protein in broken cells supernatant after induction cultivation for 12 h and 24 h, lanes 2 and 7, non optimized pGH gene for the expression of insoluble pGH protein in whole cells after induction cultivation for 12 h and 24 h, lanes 3 and 8, optimized pGH gene for the expression of soluble pGH protein in broken cells supernatant after induction cultivation for 12 h and 24 h, lanes 4 and 9, optimized pGH gene for the expression of insoluble pGH protein in whole cells after induction cultivation for 12 h and 24 h, lane M, the standard molecular weight Marker of 3452 wide type protein, lane 5, Pichia pastoris X33 of control sample. b Western-blot analysis of pGH protein of Pichia pastoris X33 expressing optimized pGH under the control of AOX1 promoter after induction at 30 °C for 24 h. lane 1, optimized pGH gene for the expression of insoluble pGH protein in broken cells sedimentation, lane 2, non optimized pGH gene for the expression of insoluble pGH protein in broken cells sedimentation, lane 3, Pichia pastoris X33 of control sample, lane 4, optimized pGH gene for the expression of soluble pGH protein in broken cells supernatant, lane 5, non optimized pGH gene for the expression of soluble pGH protein in broken cells supernatant, lane M, Dual Color standard molecular weight pre dye Marker, lane 6, optimized pGH gene for the expression of total pGH protein in whole cells, lane 7, non optimized pGH gene for the expression of total pGH protein in whole cells