Fig. 5

Detection of co-expression of pGH with Ssa1–Sis1 in Pichia pastoris.a SDS–PAGE analysis of pGH protein from Pichia pastoris X33 expressing optimized pGH alone and with Ssa1–Sis1 co-expression. Lanes 1–4, X33-pPICZA-optimized pGH (pGH1) for the expression of soluble pGH protein in broken cells supernatant cultivation for 48 h, 36 h, 24 h, and 12 h, lane 5, Pichia pastoris X33 of control sample, lane M, the standard molecular weight Marker of 3452 wide type protein, lanes 6–9, X33-pPICZA-optimized pGH–Ssa1–Sis1 (pGH1–Ssa1/Sis1) for the expression of soluble pGH protein in broken cells supernatant cultivation for 12 h, 24 h, 36 h, and 48 h, lane 10, Pichia pastoris X33 of control sample, lanes 11 and 12, pGH1 and pGH1–Ssa1/Sis1 for the expression of total pGH protein in whole cells cultivation for 48 h. b Western-blot analysis of intracellular soluble pGH protein fractions from Pichia pastoris X33 expressing optimized pGH alone and with Ssa1–Sis1 co-expression. Lanes 1–4, X33-pPICZA-optimized pGH–Ssa1–Sis1 (pGH1–Ssa1/Sis1) for the expression of soluble pGH protein in broken cells supernatant cultivation for 12 h, 24 h, 36 h, and 48 h, lane M, Dual Color standard molecular weight pre dye Marker, lane 5, Pichia pastoris X33 of control sample, lanes 6–9, X33-pPICZA-optimized pGH (pGH1) for the expression of soluble pGH protein in broken cells supernatant cultivation for 12 h, 24 h, 36 h, and 48 h