Methanol addition after glucose depletion improves rPOXA 1B production under the pGap in P. pastoris X33: breaking the habit

Abstract The purpose of this study was to demonstrate that methanol addition after glucose depletion has a positive effect on improving rPOXA 1B production under the control pGap in P. pastoris. Four different culture media (A, B, C and D) were used pastoris X33/ pGapZα A- LaccPost-Stop (clone 1), containing previously optimized POXA synthetic gene coding for ostreatus laccase, which supplemented or not with methanol. Enzyme activity without influenced by components, presenting 1254.30 ± 182.44, 1373.70 1343.50 40.30 8771.61 218.79 U L −1 , respectively. In contrast, same 24 h attained 4280.43 148.82, 3339.02 64.36, 3569.39 68.38 14,868.06 461.58 at 192 h, respectively, representing an increase approximately 3.9-, 2.4-, 3.3- 1.6-fold compared Methanol supplementation had greater impact volumetric enzyme comparison biomass production. We demonstrated what theoretically biochemically expected: recombinant protein successful, as feasible laboratory strategy sequential carbon source addition, breaking habit utilizing glucose.